Preparation and application of pyrimido thiapyrone KRAS mutant protein inhibitor

Abstract

The invention relates to a pyrimido thiapyrone KRAS mutant protein inhibitor. The pyrimido thiapyrone KRAS mutant protein inhibitor has good activity of inhibiting tumor growth. And the safety is good. The invention discloses preparation and application of a Kras inhibitor, and particularly provides a compound shown in a formula (I) and a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound and / or the pharmaceutically acceptable salt thereof, and application of the compound or the pharmaceutically acceptable salt in drugs for treating or preventing Kras kinase related diseases, especially tumors. The invention relates to a heterocyclic compound, and discloses a preparation method of a pharmaceutical composition of the compound or a pharmaceutically acceptable salt thereof. Wherein each substituent in the general formula (I) is as defined in the specification. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical synthesis, and particularly relates to a novel KRAS inhibitor and its preparation method and use.

[0002]

Prior Art

[0003] The present invention generally relates to new compounds and their preparation methods and uses as KRAS inhibitors (for example, for the treatment of cancer).

[0004] RAS represents a group of closely related monomeric globular proteins of 189 amino acids (molecular weight 21 kDa), which are associated with the plasma membrane and bind GDP or GTP. RAS acts as a molecular switch. When RAS contains bound GDP, it is in a resting or closed state and is in an "inactive state". In response to the cell's exposure to certain growth-promoting stimuli, RAS is induced to convert its bound GDP into GTP. After binding to GTP, RAS is "turned on" and is able to interact with other proteins (its "downstream targets") and initiate other proteins. The RAS protein itself has a very low intrinsic ability to hydrolyze GTP back to GDP, thus keeping itself in a closed state. Turning off RAS requires extrinsic proteins called GTPase-activating proteins (GAPs), which interact with RAS and greatly accelerate 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 an extended activation time of the protein, leading to extended cell signaling and allowing it to continue growing and dividing. Since these signals cause cell growth and division, overactive RAS signaling may ultimately lead to cancer.

[0005] Structurally, the RAS protein contains a G domain that is responsible for the enzymatic activity of RAS - guanine nucleotide binding and hydrolysis (the GTPase reaction). It also contains a C - terminal extension called the CAAX box, which undergoes post - translational modification and is responsible for targeting the protein to the membrane. The G domain is approximately 21 - 25 kDa in size and it contains a phosphate - binding loop (P - loop). The P - loop is the pocket for nucleotide binding in the protein and is a rigid part of the domain with conserved amino acid residues ((glycine 12, threonine 26, and lysine 16)), which is crucial 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 the "spring - loaded" mechanism because they are able to switch between a resting and a loaded state. The key interaction is a hydrogen bond formed by threonine 35 and glycine 60 with the γ - phosphate of GTP, which keeps the Switch1 and Switch2 regions in their active conformations respectively. After GTP hydrolysis and release of the phosphate, these two relax to an inactive GDP conformation.

[0006] The best - known members of the RAS subfamily are HRAS, KRAS, and NRAS, mainly because they are associated with various types of cancer. Mutations in any of the three major isoform (HRAS, NRAS, or KRAS) genes of RAS are among the most common in human tumorigenesis. It is found that approximately 30% of human tumors carry RAS gene mutations. Notably, KRAS mutations are detected in 25 - 30% of tumors. In contrast, the rate of oncogenic mutations occurring 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 in the P - loop and at residue Q61. G12C is a frequent mutation of the KRAS gene (glycine 12 mutated to cysteine). This mutation has been found in approximately 13% of cancers, approximately 43% of lung cancers, and approximately 100% of MYH - associated polyposis (familial colon cancer syndrome).

[0007] As a cutting-edge target, the KRAS mutant protein has received extensive attention. Among them, AMG-510 developed by Amgen was approved by the FDA for marketing 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, although progress has been made in this field, there is still a need in the art for improved compounds and methods for treating cancer, such as treating cancer by inhibiting KRAS, HRAS, or NRAS. The present invention meets this need and provides other related advantages.

[0008] In short, the present invention provides compounds capable of inhibiting KRAS mutations, including their stereoisomers, pharmaceutically acceptable salts, tautomers, and prodrugs. Summary of the Invention

[0009] A compound having the general formula (I), its stereoisomers, pharmaceutically acceptable salts, or isomers, wherein the structure of the compound represented by the general formula (I) is as follows:

[0010]

[0011] Each L1, each time it appears, is independently selected from a bond, OC 0-6 alkyl, NHC 0-6 alkyl, C 1-6 alkyl, COC 0-6 alkyl or SC 0-6 alkyl;

[0012] Each Ar, each time it appears, is independently selected from a 5- to 12-membered heteroaryl group, and the heteroaryl group independently contains 1, 2, 3, or 4 heteroatoms selected from N, O, or S each time it appears, wherein the heteroaryl group is optionally substituted by one or more G 1 substituted;

[0013] Each X1, each time it appears, is independently selected from N, CR4;

[0014] Each R2, R3, R4 is independently selected from H, D, cyano, halogen, C 1-6 alkyl, CN;

[0015] Each R1 is independently selected from H, D, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, CN, C 3-6Carbocyclic group, 3- to 10-membered heterocyclic ring, 4- to 10-membered hetero-fused ring, 5- to 12-membered spiroheterocyclic group; the 3- to 10-membered heterocyclic ring, 4- to 10-membered hetero-fused ring, 5- to 12-membered spiroheterocyclic group each independently contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S each time it appears; each R1 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 carbocyclic group, 3- to 10-membered heterocyclic ring, either substituted or unsubstituted;

[0016] U is selected from 3- to 8-membered cycloalkyl, 3- to 8-membered heteroalkyl, 5- to 12-membered fused alkyl, 5- to 12-membered fused heteroaryl, 5- to 12-membered spirocyclic group, 5- to 12-membered spiroheterocyclic group, aryl or heteroaryl, each heteroalkyl, fused heteroaryl, spiroheterocyclic group, heteroaryl independently contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S each time it appears, wherein the cycloalkyl, heteroalkyl, spirocyclic group, fused ring group, fused heteroaryl, spiroheterocyclic group, aryl or heteroaryl is optionally substituted with one or more G 2 substituted;

[0017] G 1 and G 2 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- to 8-membered heterocyclic ring, C 6-10 aryl, 5- to 10-membered heteroaryl, -OR5, -OC(O)NR5R6, -C(O)OR5, -C(O)NR5R6, -C(O)R5, -NR5R6, -NR5C(O)R6, -NR5C(O)NR6R7, -S(O) i R5 or -NR5S(O) i R6, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, aryl, heteroaryl is optionally substituted with one or more deuterium, cyano, halogen, C 1-7 alkyl, C 2-7 alkenyl, C 2-7 alkynyl, C 3-9 cycloalkyl or 3- to 9-membered heterocyclic ring, C 7-10 aryl, 6- to 10-membered heteroaryl, -OR8, -OC(O)NR8R9, -C(O)OR8, -C(O)NR8R9, -C(O)R8, -NR8R9, -NR8C(O)R9, -NR8C(O)NR9R 10 and -S(O) i R8 or -NR8S(O) isubstituted by the substituents of R9;

[0018] R5, R6, R7, R8, R9 and R 10 are each independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 alkyl, C 3-8 cycloalkyl or a 3- to 8-membered monocyclic heterocyclic group, monocyclic heteroaryl or phenyl;

[0019] m and i are 1 or 2.

[0020] In some embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its stereoisomer, formula (I) is further represented by (II-A), (II-B), (II-C) or (II-D)

[0021]

[0022]

[0023] In some embodiments, the compound of formula (I), its pharmaceutically acceptable salt or its stereoisomer, formula (I) is further represented by (II-A), (II-B), (II-C), (II-D), (II-E) or (II-F)

[0024]

[0025] In some embodiments, the compound of formula (I) or its stereoisomer, solvate or precursor, or their pharmaceutically acceptable salts are selected from the following compounds, their isomers, solvates or precursors, or their pharmaceutically acceptable salts:

[0026]

[0027]

[0028]

[0029]

[0030] In some embodiments, the compound of formula (I) or its stereoisomer, solvate or precursor, or their pharmaceutically acceptable salts are selected from the following compounds, their isomers, solvates or precursors, or their pharmaceutically acceptable salts:

[0031]

[0032]

[0033]

[0034]

[0035]

[0036] On the other hand, the present invention also provides a pharmaceutical composition, which comprises the compounds shown by formula (I) and formula (II) or their pharmaceutically acceptable salts and pharmaceutically acceptable excipients.

[0037] On the other hand, the present invention relates to a method for treating diseases related to KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H in mammals, including administering to a mammal in need of such treatment, preferably a human, a therapeutically effective amount of the compounds shown by formula (I) and formula (II) or their pharmaceutically acceptable salts, or their pharmaceutical compositions.

[0038] On the other hand, the present invention relates to the use of the compounds shown by formula (I) and formula (II) or their pharmaceutically acceptable salts in the preparation of a medicament 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.

[0039] On the other hand, the present invention relates to the compounds shown by formula (I) and formula (II) or their pharmaceutically acceptable salts, or their pharmaceutical compositions 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.

[0040] Certain chemical terms

[0041] Unless otherwise stated, the following terms used in the specification and claims.

[0042] The expression "C x-y " used herein represents a range of carbon atom numbers, where x and y are both integers. For example, C 3-8 Cycloalkyl represents a cycloalkyl having 3 - 8 carbon atoms, that is, 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 - ranges therein. For example, C 3-7 , C 3-6 , C 4-7 , C 4-6, C 5-6 etc.

[0043] "Alkyl" refers to a straight-chain or branched hydrocarbon 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.

[0044] "Alkenyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon double bond and usually 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 may be substituted or unsubstituted.

[0045] "Alkynyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond and usually 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.

[0046] "Cycloalkyl" refers to a saturated cyclic hydrocarbon group substituent containing 3 to 14 carbon ring atoms. The cycloalkyl may be a monocyclic ring, usually containing 3 to 7 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Optionally, the cycloalkyl may be a bicyclic or tricyclic ring fused together, such as decahydronaphthyl. The cycloalkyl may be substituted or unsubstituted.

[0047] "Heterocyclic group", "heterocycloalkyl group", and "heterocycle" refer to stable 3- to 18-membered monovalent non-aromatic rings, including 2-12 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, heterocyclic groups can be monocyclic, bicyclic, tricyclic, or tetracyclic systems, which may contain fused rings, spiro rings, or bridged ring systems. 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 via a carbon atom or a heteroatom on the ring. A heterocyclic group containing a fused ring can contain one or more aromatic rings or heteroaromatic rings, provided that the atom connecting to the rest of the molecule is on a non-aromatic ring. For the purposes of this application, the heterocyclic group is preferably a stable 4- to 11-membered monovalent non-aromatic monocyclic or bicyclic ring, which contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, and more preferably a stable 4- to 8-membered monovalent non-aromatic monocyclic ring, which contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heterocyclic groups include azepanyl, azetidinyl, decahydroisoquinolinyl, dihydrofuranyl, dihydroindolyl, dioxolanyl, 1,1-dioxo-thiomorpholinyl, imidazolidinyl, imidazolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidinone, pyranyl, pyrazolidinyl, pyrrolidinyl, quinazinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydropyranyl, etc.

[0048] "Spiroheterocyclic group" refers to a polycyclic heterocyclic group having 5 to 20 members, where a single atom (referred to as a spiro atom) is shared between monocyclic rings, and one or more of the ring atoms are selected from nitrogen, oxygen, or S(O) m (where m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. These can contain one or more double bonds, but none of the rings have a completely conjugated electron system. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. Spirolalkyl groups are classified into monospiroheterocyclic groups, bisspiroheterocyclic groups, or multispiroheterocyclic groups according to the number of spiro atoms shared between rings, and monospiroalkyl groups and bisspiroalkyl groups are preferred. More preferably, they are 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospiro groups. Non-limiting examples of spiroheterocyclic groups include:

[0049]

[0050] "Fused heterocyclic group" refers to a polycyclic heterocyclic group having 5 to 20 members, where each ring in the system shares an adjacent pair of atoms with other rings in the system, and one or more of the rings can contain one or more double bonds, but none of the rings have a completely conjugated π electron system, and one or more of the ring atoms are selected from nitrogen, oxygen, or S(O) ma heteroatom (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. It is preferably 6 to 14-membered, more preferably 7 to 10-membered. According to the number of rings forming the ring, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of the fused heterocyclic group include:

[0051]

[0052] "Aryl" or "aromatic group" refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10-membered, such as phenyl and naphthyl, more preferably phenyl. The aryl ring can be fused to a heteroaryl, heterocyclic or cycloalkyl ring, and the ring connected to the parent structure is the aryl ring.

[0053] "Heteroaryl" or "heteroaromatic group" refers to a 5-16 membered cyclic system that contains 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 can be a monocyclic, bicyclic, tricyclic or tetracyclic system, which may contain fused rings or bridged ring systems, as long as the connection point to the other part of the molecule is an aromatic ring atom. The nitrogen, carbon and sulfur atoms on the heteroaromatic ring can be selectively oxidized, and the nitrogen atom can be selectively quaternized. For the purposes of the present invention, the heteroaryl is preferably a stable 4-11 membered monocyclic aromatic ring that contains 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 5-8 membered monocyclic aromatic ring that contains 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of the heteroaryl include acridinyl, azepinyl, benzimidazolyl, benzindolyl, benzodioxolyl, benzodioxolyl, benzofuranone, benzofuranyl, benzonaphthofuranyl, benzopyrone, benzopyranyl, benzopyrazolyl, benzothiadiazolyl, benzothiazolyl, benzotriazolyl, furyl, imidazolyl, indazolyl, indolyl, oxazolyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinuclidinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, etc. In the present application, the heteroaryl is preferably a 5-8 membered heteroaryl that contains 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably pyridyl, pyrimidinyl, thiazolyl. The heteroaryl can be substituted or unsubstituted.

[0054] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0055] "Hydroxy" refers to -OH, "amino" refers to -NH2, "amide group" refers to -NHCO-, "cyano" refers to -CN, "nitro" refers to -CN, "isocyano" refers to -NC, "trifluoromethyl" refers to -CF3.

[0056] As used herein, the term "heteroatom" or "hetero" when used alone or as part of other components refers to an atom other than carbon and hydrogen. The heteroatoms 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 heteroatoms are present, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different from each other.

[0057] As used herein, the term "fused" or "fused ring" when used alone or in combination refers to a cyclic structure in which two or more rings share one or more bonds.

[0058] As used herein, the term "spiro" or "spiro ring" when used alone or in combination refers to a cyclic structure in which two or more rings share one or more atoms.

[0059] "Optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and this description includes the instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted by an alkyl group" means that the alkyl group may, but need not, be present, and this description includes the cases where the heterocyclic group is substituted by an alkyl group and the cases where the heterocyclic group is not substituted by an alkyl group.

[0060] "Substituted" means that one or more atoms in a group, preferably 5 atoms, more preferably 1 to 3 atoms, are independently replaced by a corresponding number of substituents. It goes without saying that the substituents are at their possible chemical positions, and those skilled in the art can determine (by experiment or theory) the possible or impossible substitutions without undue effort. For example, a free amino group or hydroxyl group may be unstable when combined with a carbon atom having an unsaturated (such as an olefin) bond. The substituents include, but are not limited to, hydroxyl, amino, halogen, cyano, C 1-6 alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, and the like.

[0061] "Pharmaceutical composition" refers to a composition containing one or more of the compounds described herein or their pharmaceutically acceptable salts or prodrugs, as well as other components, such as pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate the administration to an organism, facilitate the absorption of the active ingredient, and thereby exert biological activity.

[0062] "Isomers" refer to compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or in the spatial arrangement of their atoms. Isomers with different spatial arrangements of their atoms are called "stereoisomers". Stereoisomers include optical isomers, geometric isomers, and conformational isomers. The compounds of the present invention can exist in the form of optical isomers. Depending on the configuration of the substituents around the chiral carbon atom, these optical isomers are of the "R" or "S" configuration. Optical isomers include enantiomers and diastereomers. Methods for preparing and separating optical isomers are known in the art.

[0063] The compounds of the present invention can also exist as 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 groups, or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as the Z or E configuration, and substituents around cycloalkyl or heterocyclic groups are designated as the cis or trans configuration.

[0064] The compounds of the present invention may also exhibit tautomerism, such as keto-enol tautomerism.

[0065] 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 structural formula of the compound.

[0066] "Isotopes" are all isotopes of atoms that occur in the compounds of the present invention. Isotopes include those atoms that have 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 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 31 P, 32 P, 35 S, 18 F and 36 Cl. Isotopically 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 appended examples using appropriate isotopically labeled reagents in place of non-isotopically labeled reagents. Such compounds have various potential uses, such as as standards and reagents in the determination of biological activity. In the case of stable isotopes, such compounds have the potential to favorably alter biological, pharmacological, or pharmacokinetic properties.

[0067] "Prodrug" means that the compounds of the present invention can be administered in the form of prodrugs. A prodrug is a derivative that is converted into the bioactive compound of the present invention under physiological conditions in vivo, such as by oxidation, reduction, hydrolysis, etc. (each of which utilizes enzymes or proceeds without the participation of enzymes). Examples of prodrugs are the following compounds: wherein the amino group in the compound of the present invention is acylated, alkylated or phosphorylated, such as eicosanoylamino group, propionamide group, pivaloyloxymethylamino group, or wherein the hydroxyl group is acylated, alkylated, phosphorylated or converted into borate, such as acetoxy group, palmitoyloxy group, pivaloyloxy group, succinyloxy group, fumaroyloxy group, propionoyloxy group, or wherein the carboxyl group is esterified or amidated, or wherein the mercapto group forms a disulfide bridge with a carrier molecule that selectively delivers the drug to the target and / or to the cytosol of the cell, such as a peptide. These compounds can be prepared from the compounds of the present invention according to known methods.

[0068] "Pharmaceutically acceptable salts" or "pharmaceutically acceptable" means those made from pharmaceutically acceptable bases or acids, including inorganic or organic bases or acids. In the case where the compounds of the present invention contain one or more acidic or basic groups, the present invention also encompasses their corresponding pharmaceutically acceptable salts. Thus, the compounds of the present invention containing acidic groups can exist in the form of salts and can be used according to the present invention, for example, as alkali metal salts, alkaline earth metal salts or as ammonium salts. More specific examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts or salts 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 their addition with inorganic or organic acids. Examples of suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, phosphorous acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic 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, malic 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 inner salts or inner ammonium salts in addition to the salt forms mentioned above. Each salt is obtained by conventional methods known to those skilled in the art, such as by contacting these with organic or inorganic acids or bases in a solvent or dispersant or by anion exchange or cation exchange with other salts.

[0069] Accordingly, as used in this application, when referring to "compound", "compound of the present invention" or "compounds of the present invention", it includes all forms of said compounds, such as their prodrugs, stable isotope derivatives, pharmaceutically acceptable salts, isomers, meso forms, racemates, enantiomers, diastereomers and mixtures thereof.

[0070] As used herein, the term "tumor" includes benign tumors and malignant tumors (such as cancer).

[0071] As used herein, the term "cancer" includes various malignant tumors in which KRAS is involved in its occurrence, 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 granular cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, throat cancer, breast cancer, prostate cancer, glioblastoma, 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, splenomegalic erythrocytosis, eosinophilic leukocytosis syndrome, multiple myeloma and other various solid tumors and hematological tumors.

[0072] As used herein, the term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" refers to the amount of at least one agent or compound that, when administered, is sufficient to relieve to some extent one or more symptoms of the disease or disorder being treated. The result can be the reduction and / or alleviation of signs, symptoms or causative factors and / or any other desired change in the biological system. For example, the "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is required to provide a significant alleviation of the disorder clinically. Techniques such as dose escalation trials can be used to determine the effective amount suitable for any individual case.

[0073] As used in the present invention, the term "polymorph" or "polymorphism" means that the compounds of the present invention have multiple crystal lattice forms, and some compounds of the present invention may have more than one crystal form. The present invention encompasses all polymorphic forms or mixtures thereof.

[0074] Intermediate compounds of the compounds of the present invention and their polymorphs are also within the scope of the present invention.

[0075] Crystallization often produces solvates of the compounds of the present invention. As used herein, the term "solvate" refers to an entity formed by the combination of one or more molecules of a compound of the present invention and one or more solvent molecules.

[0076] The solvent can be water, in which case the solvate is a hydrate. Additionally, it can also be an organic solvent. Therefore, the compounds of the present invention can exist as hydrates, including monohydrates, dihydrates, hemihydrates, trihydrates, tetrahydrates, etc., as well as the corresponding solvated forms. The compounds of the present invention can be true solvates, but in some other cases, the compounds of the present invention may only accidentally retain water or a mixture of water and some other solvents. The compounds of the present invention can react in a solvent or precipitate or crystallize in a solvent. The solvates of the compounds of the present invention are also included within the scope of the present invention.

[0077] As used herein, the term "acceptable" in relation to a formulation, composition, or ingredient means that it has no continuing adverse effect on the overall health of the treated subject.

[0078] As used herein, the term "pharmaceutically acceptable" 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 an individual without causing an adverse biological reaction or interacting in an adverse manner with any of the components contained in the composition.

[0079] "Pharmaceutically acceptable carriers" include, but are not limited to, adjuvants, carriers, excipients, auxiliaries, deodorants, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants and wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that have been approved by the relevant government administrative departments for use in humans and domesticated animals.

[0080] As used herein, the terms "subject", "patient", "object", or "individual" refer to an individual 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 (such as chimpanzees and other apes and monkeys); domestic animals, such as cows, horses, sheep, goats, pigs; household 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.

[0081] As used herein, the term "treatment" refers to the treatment of a relevant disease or disorder in a mammal, especially a human, including

[0082] (i) preventing a mammal, especially a mammal that has been previously exposed to a disease or disorder but has not been diagnosed with the disease or disorder, from developing the corresponding disease or disorder;

[0083] (ii) inhibiting a disease or disorder, i.e., controlling its development;

[0084] (iii) Alleviating a disease or disorder, i.e., causing the disease or disorder to subside slowly;

[0085] (iv) Alleviating the symptoms caused by a disease or disorder.

[0086] As used herein, the terms "disease" and "disorder" may be used interchangeably or may have different meanings. Since certain specific diseases or disorders do not yet have known causative agents (so the cause of the disease is still unclear), they cannot yet be recognized as diseases but can only be regarded as unwanted conditions or syndromes, and more or less specific symptoms of these syndromes have been confirmed by clinical researchers.

[0087] As used herein, the terms "administering", "applying", "dosing", etc. refer to methods that can deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral route, duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injection or infusion), topical administration, and rectal administration. In a preferred embodiment, the compounds and compositions discussed herein are administered orally. Specific implementation methods

[0089] The present invention also provides a method for preparing the compound. The preparation of the compound of 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 compounds of the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or by combining known methods in the art and the methods of the present invention. The product obtained in each step should be 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 synthesized conventionally according to the literature (Reaxys) or purchased.

[0090] Unless otherwise specified, the temperature is in degrees Celsius. The reagents are purchased from commercial suppliers such as Chemblocks Inc and 3wpharm, and these reagents can be used directly without further purification, unless otherwise specified.

[0091] Unless otherwise specified, the following reactions are carried out at room temperature, in an anhydrous solvent, under a positive pressure of nitrogen or argon gas, or using a drying tube; the glassware is dried by baking and / or heating.

[0092] Unless otherwise specified, column chromatography purification uses silica gel with a mesh size of 200 - 300 from Qingdao Ocean Chemical Factory; preparative thin-layer chromatography uses prefabricated thin-layer chromatography silica gel plates (HSGF254) produced by Yantai Chemical Industry Research Institute; the determination of MS is carried out using a Thermo LCD Fleet type (ESI) liquid chromatography - mass spectrometry instrument.

[0093] Nuclear magnetic data (1H NMR) was obtained using a Bruker Avance-400 MHz or Varian Oxford-400 Hz nuclear magnetic resonance spectrometer. The solvents used for nuclear magnetic data were CDCl3, CD3OD, D2O, DMS-d6, etc. Tetramethylsilane (0.000 ppm) or the residual solvent was used as the reference (CDCl 3: 7.26 ppm; CD3OD: 3.31 ppm; D2O: 4.79 ppm; d6-DMSO: 2.50 ppm). When indicating the multiplicity of peaks, the following abbreviations are used to represent different peak multiplicities: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad peak), dd (doublet of doublets), dt (doublet of triplets). If coupling constants are given, they are in Hertz (Hz).

[0094] Preparation of intermediates

[0095] Preparation of A1

[0096]

[0097] Step A

[0098] In a dry 1 L three-necked flask, potassium carbonate (27.6 g, 200 mmol) was added to N,N-dimethylformamide (100 mL). The reaction system was a heterogeneous gray color. The temperature was lowered to 0 °C, and a solution of compound A1-1 (10 g, 80 mmol) in N,N-dimethylformamide (200 mL) was added dropwise under nitrogen protection. The reaction was continued at 0 °C for 0.5 h. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, and the temperature was slowly raised to 20 °C and stirring was continued under nitrogen protection for 7.5 h. The reaction solution was slowly added to 100 mL of saturated ammonium chloride, extracted with methyl tert-butyl ether (100 mL × 2), and the combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product obtained was separated and purified by silica gel column chromatography to obtain compound A1-2. LC-MS (ESI): m / z = 365.45 [M+H] + 。

[0099] Step B

[0100] 2,2,6,6 - Tetramethylpiperidine (11.4 g, 82.1 mmol) was added to anhydrous tetrahydrofuran (500 mL). The temperature was lowered to -5 °C, and n-butyllithium (2.5 M, 40 mL) was added dropwise. The reaction was carried out at -5 to 0 °C for 15 minutes, then the temperature was lowered to -60 °C. A solution of compound A1-2 (10 g, 27 mmol) in tetrahydrofuran (30 mL) was added, and the reaction was carried out at -60 °C for 0.5 hour. N,N-Dimethylformamide (40 g, 0.5 mol) was quickly added, and the reaction mixture was stirred at -60 °C for an additional 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction mixture, and the mixture was extracted with methyl tert-butyl ether (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a solvent for 0.5 hour, filtered, the filter cake was dried, and the filtrate was concentrated and purified by silica gel column chromatography. The filter cake and the column chromatography fractions were combined to obtain compound A1-3. LC-MS (ESI): m / z = 393.5 [M+H] + 。

[0101] Step C

[0102] Compound A1-3 (10 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and N-bromosuccinimide (4.5 g, 25 mmol) was added. The reaction mixture was stirred at 20 °C for an additional 20 minutes. The reaction mixture was added to 150 mL of water, and the mixture was extracted with methyl tert-butyl ether (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a mixed solvent for 0.5 hour, filtered, and the filter cake was dried to obtain compound A1-4. LC-MS (ESI): m / z = 472.4 [M+H] + 。

[0103] Step D

[0104] Compound A1-4 (9.7 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Under nitrogen, copper(I) iodide (7.8 g, 41 mmol) and methyl fluorosulfonyldifluoroacetate (20 g, 100 mmol) were added to the reaction mixture. The reaction mixture was heated to 100 °C and stirred for 1 hour. After cooling, the reaction mixture was filtered through diatomaceous earth. The filtrate was added to 150 mL of water, and the mixture was extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to obtain compound A1-5. LC-MS (ESI): m / z = 461.5 [M+H] + 。

[0105] Step E

[0106] Under nitrogen protection, A1-5 (5.98 g, 13 mmol) and lithium hydroxide (0.82 g, 19.5 mmol) were dissolved in 100 mL of THF at room temperature. Then, dimethyl ethylphosphonoacetate (3.55 g, 19.5 mmol) was added, and the reaction mixture was stirred at room temperature for 24 hours. THF was recovered, and the mixture was added to 150 mL of water. It was extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound A1-6. LC-MS (ESI): m / z = 531.6 [M+H] + 。

[0107] Step F

[0108] Compound A1-6 (10.6 g, 20 mmol) and piperidine (0.2 mL) were added to dichloromethane (100 mL). Ethyl 3-mercaptopropionate (3.2 g, 24 mmol) and benzyltrimethylammonium hydroxide (1 mL, 30% methanol) were added, and the mixture was stirred at 60 °C for 48 hours. Dilute hydrochloric acid was added, and it was extracted from dichloromethane (75 mL × 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound A1-7. LC-MS (ESI): m / z = 665.7 [M+H] + 。

[0109] Step G

[0110] Compound A1-7 (7.98 g, 12 mmol) and sodium ethoxide (1.36 g, 19 mmol) were dissolved in 400 mL of toluene, and the mixture was stirred at room temperature for 24 hours. It was poured into 100 mL of water and extracted from dichloromethane (100 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound A1-8. LC-MS (ESI): m / z = 619.7 [M+H] + 。

[0111] Step H

[0112] To a suspension of A1-8 (3.3 g, 5.3 mmol) in water (10 mL), S-methylisothiourea sulfate (1.7 g, 6.2 mmol) and potassium carbonate (1.6 g, 11.7 mmol) were added at room temperature. The reaction mixture was stirred for 20 h. The precipitate was collected, washed with water (twice) and isopropyl ether, dried, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound A1-9. LC-MS (ESI): m / z = 645.7 [M+H] + 。

[0113] Step I

[0114] Compound A1-9 (1 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), N,N-diisopropylethylamine (0.6 g, 4.8 mmol) was added, the temperature was lowered to 0 - 10 °C, and trifluoromethanesulfonic anhydride (0.7 g, 2.4 mmol, 400 μL) was slowly added to the reaction solution. The reaction was carried out at this temperature for 15 min. The reaction solution was poured into saturated ammonium chloride aqueous solution (8 mL), liquid separation was carried out, the aqueous phase was extracted with dichloromethane (5 mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound A1. LC-MS (ESI): m / z = 777.8 [M+H] + 。

[0115] Preparation of B1

[0116]

[0117] Step A

[0118] To a solution of B1-1 (25.3 g, 201 mmol) in N,N-dimethylformamide (500 mL), bromomethylbenzene (41.2 g, 241 mmol) and potassium carbonate (55.5 g, 402 mmol) were added. After stirring at 25 °C for 15 h, the reaction mixture was diluted with ethyl acetate and washed with water and brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. It was purified by silica gel column chromatography to obtain B1-2. LC-MS (ESI): m / z = 216.3 [M+H] + 。

[0119] Step B

[0120] 2,2,6,6 - Tetramethylpiperidine (11.4 g, 82.1 mmol) was added to anhydrous tetrahydrofuran (500 mL). The temperature was lowered to -5 °C, and n-butyllithium (2.5 M, 40 mL) was added dropwise. The reaction was carried out at -5 to 0 °C for 15 minutes. Then the temperature was lowered to -60 °C, and a solution of compound B1-2 (5.8 g, 27 mmol) in tetrahydrofuran (30 mL) was added. The reaction was carried out at -60 °C for 0.5 hour. N,N-Dimethylformamide (40 g, 0.5 mol) was quickly added, and the reaction mixture was stirred at -60 °C for an additional 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction mixture, and it was extracted with methyl tert-butyl ether (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a solvent for 0.5 hour, filtered, the filter cake was dried, and the filtrate was concentrated and then purified by silica gel column chromatography. The filter cake and the column chromatography fractions were combined to obtain compound B1-3. LC-MS (ESI): m / z = 245.1 [M + H] + 。

[0121] Step C

[0122] Compound B1-3 (6.1 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and N-bromosuccinimide (4.5 g, 25 mmol) was added. The reaction mixture was stirred at 20 °C for an additional 20 minutes. The reaction mixture was added to 150 mL of water and extracted with methyl tert-butyl ether (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a mixed solvent for 0.5 hour, filtered, and the filter cake was dried to obtain compound B1-4. LC-MS (ESI): m / z = 323.2 [M + H] + 。

[0123] Step D

[0124] Compound B1-4 (6.57 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Under nitrogen, cuprous iodide (7.8 g, 41 mmol) and methyl fluorosulfonyldifluoroacetate (20 g, 100 mmol) were added to the reaction mixture. The reaction mixture was heated to 100 °C and stirred for 1 hour. After cooling, the reaction mixture was filtered through diatomaceous earth. The filtrate was added to 150 mL of water and extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to obtain compound B1-5. LC-MS (ESI): m / z = 313.3 [M + H] + 。

[0125] Step E

[0126] Under nitrogen protection, B1-5 (4.06 g, 13 mmol) and lithium hydroxide (0.82 g, 19.5 mmol) were dissolved in 100 mL of THF at room temperature, then dimethyl ethylphosphonoacetate (3.55 g, 19.5 mmol) was added, and the reaction solution was stirred at room temperature for 24 hours. THF was recovered, added to 150 mL of water, extracted with methyl tert-butyl ether (75 mL × 2), the combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound B1-6. LC-MS (ESI): m / z = 383.2 [M+H] + 。

[0127] Step F

[0128] Compound B1-6 (7.64 g, 20 mmol) and piperidine (0.2 mL) were added to dichloromethane (100 mL), ethyl 3-mercaptopropionate (3.2 g, 24 mmol) and benzyltrimethylammonium hydroxide (1 mL, 30% methanol) were added, and the reaction was stirred at 60 °C for 48 hours. Dilute hydrochloric acid was added, extracted from dichloromethane (75 mL × 2), the combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound B1-7. LC-MS (ESI): m / z = 517.2 [M+H] + 。

[0129] Step G

[0130] Compound B1-7 (6.2 g, 12 mmol) and sodium ethoxide (1.36 g, 19 mmol) were dissolved in 400 mL of toluene, and the reaction was stirred at room temperature for 24 hours. Poured into 100 mL of water, extracted from dichloromethane (100 mL × 2), the combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound B1-8. LC-MS (ESI): m / z = 471.2 [M+H] + 。

[0131] Step H

[0132] To a suspension of B1-8 (2.49 g, 5.3 mmol) in water (10 mL), S-methylisothiourea sulfate (1.7 g, 6.2 mmol) and potassium carbonate (1.6 g, 11.7 mmol) were added at room temperature. The reaction mixture was stirred for 20 h. The precipitate was collected, washed with water (twice) and isopropyl ether, dried, and the crude product obtained was separated and purified by silica gel column chromatography to give compound B1-9. LC-MS (ESI): m / z = 497.1 [M+H] + 。

[0133] Step I

[0134] Compound B1-9 (0.79 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), N,N-diisopropylethylamine (0.6 g, 4.8 mmol) was added, the temperature was cooled to 0 - 10 °C, and trifluoromethanesulfonic anhydride (0.7 g, 2.4 mmol, 400 μL) was slowly added to the reaction solution. The reaction was carried out at this temperature for 15 min. The reaction solution was poured into saturated ammonium chloride aqueous solution (8 mL), liquid separation was carried out, the aqueous phase was extracted with dichloromethane (5 mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure, and the crude product obtained was separated and purified by silica gel column chromatography to give compound B1. LC / MS (ESI): m / z = 629.0 [M+H] + 。

[0135] Preparation of C1

[0136]

[0137] Compound C1 was obtained by a preparation method similar to that of intermediate A1 (with the raw material changed to 6-bromo-4-methylpyridin-2-amine). LC / MS (ESI): m / z = 760.8 [M+H] + 。

[0138] Preparation of D1

[0139]

[0140] Compound D1 was obtained by a preparation method similar to that of intermediate B1 (with the raw material changed to 6-bromo-4-methylpyridin-2-ol). LC / MS (ESI): m / z = 611.6 [M+H] + 。

[0141] Preparation of E1

[0142]

[0143] Step A

[0144] In a dry 1 L three-necked flask, potassium carbonate (27.6 g, 200 mmol) was added to N,N-dimethylformamide (100 mL). The temperature was lowered to 0 °C, and a solution of compound E1-1 (10 g, 80 mmol) in N,N-dimethylformamide (500 mL) was added dropwise under nitrogen protection. The reaction was continued at 0 °C for 0.5 h. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, and the temperature was slowly raised to 20 °C, and stirring was continued for 7.5 h under nitrogen protection. The reaction solution was slowly added to 100 mL of saturated ammonium chloride, extracted with methyl tert-butyl ether (100 mL × 2), the combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product obtained was separated and purified by silica gel column chromatography to obtain compound E1-2. LC-MS (ESI): m / z = 430.1 [M+H] + 。

[0145] Step B

[0146] Compound E1-2 (34.6 g, 80.7 mmol) was added to toluene (150 mL), dichlorobis[di-tert-butyl-(4-dimethylaminophenyl)phosphine]palladium(0) (0.286 g, 0.404 mmol) and tributyl(1-propynyl)tin (26.6 g, 80.7 mmol). The reaction was carried out at 120 °C for 2 h under nitrogen protection. The reaction solution was concentrated under reduced pressure and separated by column chromatography to obtain compound E1-3. LC-MS (ESI): m / z = 390 [M+H] + 。

[0147] Step C

[0148] 2,2,6,6-Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL). The temperature was lowered to -5 °C, and n-butyllithium (2.5 M, 50 mL) was added dropwise. The reaction was carried out at -5 to 0 °C for 15 min, and the temperature was lowered to -60 °C. A solution of compound E1-3 (24.27 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added, and the reaction was carried out at -60 °C for 0.5 h. N,N-Dimethylformamide (22.8 g, 0.312 mol) was added quickly, and the reaction solution was stirred at -60 °C for 10 min. 200 mL of saturated ammonium chloride was added to the reaction solution, and it was extracted with methyl tert-butyl ether (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product obtained was slurried with solvent for 0.5 h, filtered, the filter cake was dried, the filtrate was concentrated and separated and purified by silica gel column chromatography. The filter cake and the column chromatography were combined to obtain compound E1-4. LC-MS (ESI): m / z = 418.1 [M+H] + 。

[0149] Step D

[0150] Compound E1-4 (10.43 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and N-bromosuccinimide (4.5 g, 25 mmol) was added. The reaction mixture was stirred at 20 °C for an additional 20 minutes. The reaction mixture was added to 150 mL of water and extracted with methyl tert-butyl ether (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The resulting crude product was slurried in a mixed solvent for 0.5 hour, filtered, and the filter cake was dried to obtain Compound E1-5. LC-MS (ESI): m / z = 496.0 [M+H] + 。

[0151] Step E

[0152] Compound E1-5 (9.7 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Cuprous iodide (7.8 g, 4.1 mmol) and methyl fluorosulfonyldifluoroacetate (20 g, 100 mmol) were added to the reaction mixture under nitrogen. The reaction mixture was heated to 100 °C and stirred for 1 hour. After cooling, the reaction mixture was filtered through diatomaceous earth. The filtrate was added to 150 mL of water and extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography to obtain Compound E1-6. LC-MS (ESI): m / z = 496.1 [M+H] + 。

[0153] Step E

[0154] Under nitrogen protection, E1-5 (6.44 g, 13 mmol) and lithium hydroxide (0.82 g, 19.5 mmol) were dissolved in 100 mL of THF at room temperature. Then dimethyl ethylphosphonoacetate (3.55 g, 19.5 mmol) was added. The reaction mixture was stirred at room temperature for 24 hours. THF was recovered, and the mixture was added to 150 mL of water and extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography to obtain Compound E1-6. LC-MS (ESI): m / z = 556.1 [M+H] + 。

[0155] Step F

[0156] Compound E1-6 (11.1 g, 20 mmol) and piperidine (0.2 mL) were added to dichloromethane (100 mL). Ethyl 3-mercaptopropionate (3.2 g, 24 mmol) and benzyltrimethylammonium hydroxide (1 mL, 30% methanol) were added, and the mixture was stirred at 60 °C for 48 hours. Dilute hydrochloric acid was added, and the mixture was extracted with dichloromethane (75 mL × 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound E1-7. LC-MS (ESI): m / z = 690.3 [M+H] + 。

[0157] Step G

[0158] Compound E1-8 (7.97 g, 12 mmol) and sodium ethoxide (1.36 g, 19 mmol) were dissolved in toluene (400 mL), and the mixture was stirred at room temperature for 24 hours. It was poured into 100 mL of water and extracted with dichloromethane (100 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound E1-9. LC-MS (ESI): m / z =  619.7 [M+H] + 。

[0159] Step H

[0160] To a suspension of E1-9 (3.28 g, 5.3 mmol) in water (10 mL), S-methylisothiourea sulfate (1.7 g, 6.2 mmol) and potassium carbonate (1.6 g, 11.7 mmol) were added at room temperature. The reaction mixture was stirred for 20 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound E1-10. LC-MS (ESI): m / z = 644.2 [M+H] + 。

[0161] Step I

[0162] Dissolve compound E1-10 (1.03 g, 1.6 mmol) in dichloromethane (10 mL), add N,N-diisopropylethylamine (0.6 g, 4.8 mmol), cool the temperature to 0 - 10 °C, and slowly add trifluoromethanesulfonic anhydride (0.7 g, 2.4 mmol, 400 μL) to the reaction solution. React for 15 minutes at this temperature. Pour the reaction solution into saturated ammonium chloride aqueous solution (8 mL), separate the layers, extract the aqueous phase with dichloromethane (5 mL × 2), dry the combined organic phases over anhydrous sodium sulfate, filter, remove the organic solvent under reduced pressure, and purify the obtained crude product by silica gel column chromatography to obtain compound E1. LC-MS (ESI): m / z = 802.1 [M+H] + 。

[0163] Preparation of F1

[0164]

[0165] Step A

[0166] In a dry 1 L three-necked flask, add potassium carbonate (27.6 g, 200 mmol) to N,N-dimethylformamide (100 mL), cool the temperature to 0 °C, and dropwise add a solution of compound E1-1 (23.76 g, 80 mmol) in N,N-dimethylformamide (500 mL) under nitrogen protection. Continue to react at 0 °C for 0.5 hour, add p-methoxybenzyl chloride (27.5 g, 170 mmol), slowly warm up to 20 °C, and continue to stir for 7.5 hours under nitrogen protection. Slowly add the reaction solution to 100 mL of saturated ammonium chloride, extract with methyl tert-butyl ether (100 mL × 2), wash the combined organic phases with 100 mL of saturated brine, dry over anhydrous sodium sulfate, filter, remove the organic solvent under reduced pressure, and purify the obtained crude product by silica gel column chromatography to obtain compound F1-2. LC-MS (ESI): m / z = 567.9 [M+H] + 。

[0167] Step B

[0168] Add compound F1-2 (45.67 g, 80.7 mmol), toluene (150 mL), dichlorobis[di-tert-butyl-(4-dimethylaminophenyl)phosphine]palladium (0.286 g, 0.404 mmol), and tributyl(1-propynyl)tin (26.6 g, 80.7 mmol), and react at 120 °C for 2 hours under nitrogen protection. Concentrate the reaction solution under reduced pressure and separate by column chromatography to obtain compound F1-3. LC-MS (ESI): m / z = 451 [M+H] + 。

[0169] Step C

[0170] 2,2,6,6 - Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL). The temperature was lowered to -5 °C, and n-butyllithium (2.5 M, 50 mL) was added dropwise. The reaction was carried out at -5 to 0 °C for 15 minutes. Then the temperature was lowered to -60 °C, and a solution of compound E1-3 (28.08 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The reaction was carried out at -60 °C for 0.5 hour. N,N-Dimethylformamide (22.8 g, 0.312 mol) was quickly added, and the reaction mixture was stirred at -60 °C for an additional 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction mixture, and it was extracted with methyl tert-butyl ether (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a solvent for 0.5 hour, filtered, and the filter cake was dried. The filtrate was concentrated and then purified by silica gel column chromatography. The filter cake and the column chromatography fractions were combined to obtain compound F1-4. LC-MS (ESI): m / z = 400.2 [M+H] + 。

[0171] Step D

[0172] Compound F1-4 (10.43 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide. N-Bromosuccinimide (4.5 g, 25 mmol) was added, and the reaction mixture was stirred at 20 °C for 20 minutes. The reaction mixture was added to 150 mL of water and extracted with methyl tert-butyl ether (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a mixed solvent for 0.5 hour, filtered, and the filter cake was dried to obtain compound F1-5. LC-MS (ESI): m / z = 478.0 [M+H] + 。

[0173] Step E

[0174] Compound F1-5 (9.77 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Under nitrogen, cuprous iodide (7.8 g, 4.1 mmol) and methyl fluorosulfonyldifluoroacetate (2 g, 100 mmol) were added to the reaction mixture. The reaction mixture was heated to 100 °C and stirred for 1 hour. After cooling, the reaction mixture was filtered through diatomaceous earth. The filtrate was added to 150 mL of water and extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to obtain compound F1-6. LC-MS (ESI): m / z = 468.1 [M+H] + 。

[0175] Step E

[0176] Under nitrogen protection, F1-6 (6.07 g, 13 mmol) and lithium hydroxide (0.82 g, 19.5 mmol) were dissolved in 100 mL of THF at room temperature, then dimethyl ethylphosphonoacetate (3.55 g, 19.5 mmol) was added. The reaction mixture was stirred at room temperature for 24 hours. THF was recovered, and the mixture was added to 150 mL of water. It was extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound F1-7. LC-MS (ESI): m / z = 538.2 [M+H] + 。

[0177] Step F

[0178] Compound F1-7 (10.7 g, 20 mmol) and piperidine (0.2 mL) were added to dichloromethane (100 mL). Ethyl 3-mercaptopropionate (3.2 g, 24 mmol) and benzyltrimethylammonium hydroxide (1 mL, 30% in methanol) were added, and the mixture was stirred at 60 °C for 48 hours. Dilute hydrochloric acid was added, and it was extracted from dichloromethane (75 mL × 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound F1-8. LC-MS (ESI): m / z = 670.1 [M+H] + 。

[0179] Step G

[0180] Compound F1-8 (8.39 g, 12 mmol) and sodium ethoxide (1.36 g, 19 mmol) were dissolved in 400 mL of toluene, and the mixture was stirred at room temperature for 24 hours. It was poured into 100 mL of water and extracted from dichloromethane (100 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound F1-9. LC-MS (ESI): m / z = 625.2 [M+H] + 。

[0181] Step H

[0182] To a suspension of E1-9 (3.45 g, 5.3 mmol) in water (10 mL) was added S-methylisothiourea sulfate (1.7 g, 6.2 mmol) and potassium carbonate (1.6 g, 11.7 mmol) at room temperature. The reaction mixture was stirred for 20 h. The precipitate was collected, washed with water (twice) and isopropyl ether, dried, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound F1-10. LC-MS (ESI): m / z = 652.2 [M+H] + 。

[0183] Step I

[0184] Compound F1-10 (1.04 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), N,N-diisopropylethylamine (0.6 g, 4.8 mmol) was added, the temperature was lowered to 0 - 10 °C, and trifluoromethanesulfonic anhydride (0.7 g, 2.4 mmol, 400 μL) was slowly added to the reaction solution. The reaction was carried out at this temperature for 15 min. The reaction solution was poured into saturated ammonium chloride aqueous solution (8 mL), liquid separation was carried out, the aqueous phase was extracted with dichloromethane (5 mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound F1. LC-MS (ESI): m / z = 784.1 [M+H] + 。

[0185] Preparation of G1

[0186]

[0187] Step A

[0188] In a dry 1 L three-necked flask, potassium carbonate (27.6 g, 200 mmol) was added to N,N-dimethylformamide (100 mL), the temperature was lowered to 0 °C, and a solution of compound G1-1 (16.4 g, 80 mmol) in N,N-dimethylformamide (500 mL) was added dropwise under nitrogen protection. The reaction was continued at 0 °C for 0.5 h, p-methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20 °C, and stirring was continued under nitrogen protection for 7.5 h. The reaction solution was slowly added to 100 mL of saturated ammonium chloride, extracted with methyl tert-butyl ether (100 mL × 2), the combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound G1-2. LC-MS (ESI): m / z = 467.0 [M+H] + 。

[0189] Step B

[0190] 2,2,6,6 - Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL), and the temperature was lowered to -5 °C. n-Butyllithium (2.5 M, 50 mL) was added dropwise, and the reaction was carried out at -5 to 0 °C for 15 minutes. Then the temperature was lowered to -60 °C, and a solution of compound G1-2 (29.08 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The reaction was carried out at -60 °C for 0.5 hour, and then N,N-dimethylformamide (22.8 g, 0.312 mol) was quickly added. The reaction mixture was stirred at -60 °C for an additional 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction mixture, and the mixture was extracted with methyl tert-butyl ether (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a solvent for 0.5 hour, filtered, and the filter cake was dried. The filtrate was concentrated and then purified by silica gel column chromatography. The filter cake and the column chromatography fractions were combined to obtain compound G1-3. LC-MS (ESI): m / z = 396.1 [M+H] + 。

[0191] Step D

[0192] Compound G1-3 (9.88 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and N-bromosuccinimide (4.5 g, 25 mmol) was added. The reaction mixture was stirred at 20 °C for an additional 20 minutes. The reaction mixture was added to 150 mL of water, and the mixture was extracted with methyl tert-butyl ether (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a mixed solvent for 0.5 hour, filtered, and the filter cake was dried to obtain compound G1-4. LC-MS (ESI): m / z = 474.0 [M+H] + 。

[0193] Step E

[0194] Compound G1-4 (9.65 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Under nitrogen, cuprous iodide (7.8 g, 4.1 mmol) and methyl fluorosulfonyldifluoroacetate (20 g, 100 mmol) were added to the reaction mixture. The reaction mixture was heated to 100 °C and stirred for 1 hour. After cooling, the reaction mixture was filtered through diatomaceous earth. The filtrate was added to 150 mL of water, and the mixture was extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to obtain compound G1-5. LC-MS (ESI): m / z = 464.1 [M+H] + 。

[0195] Step E

[0196] Under nitrogen protection, G1-5 (6.02 g, 13 mmol) and lithium hydroxide (0.82 g, 19.5 mmol) were dissolved in 100 mL of THF at room temperature, then dimethyl ethylphosphonoacetate (3.55 g, 19.5 mmol) was added, and the reaction mixture was stirred at room temperature for 24 hours. THF was recovered, added to 150 mL of water, extracted with methyl tert-butyl ether (75 mL × 2), the combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound G1-6. LC-MS (ESI): m / z = 534.1 [M+H] + 。

[0197] Step F

[0198] Compound G1-6 (10.6 g, 20 mmol) and piperidine (0.2 mL) were added to dichloromethane (100 mL), ethyl 3-mercaptopropionate (3.2 g, 24 mmol) and benzyltrimethylammonium hydroxide (1 mL, 30% methanol) were added, and the reaction was stirred at 60 °C for 48 hours. Dilute hydrochloric acid was added, extracted from dichloromethane (75 mL × 2), the combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound G1-7. LC-MS (ESI): m / z = 669.2 [M+H] + 。

[0199] Step G

[0200] Compound G1-7 (8.02 g, 12 mmol) and sodium ethoxide (1.36 g, 19 mmol) were dissolved in 400 mL of toluene, and the reaction was stirred at room temperature for 24 hours. Poured into 100 mL of water, extracted from dichloromethane (100 mL × 2), the combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound G1-8. LC-MS (ESI): m / z = 622.1 [M+H] + 。

[0201] Step H

[0202] To a suspension of G1-8 (3.29 g, 5.3 mmol) in water (10 mL) was added S-methylisothiourea sulfate (1.7 g, 6.2 mmol) and potassium carbonate (1.6 g, 11.7 mmol) at room temperature. The reaction mixture was stirred for 20 h. The precipitate was collected, washed with water (twice) and isopropyl ether, dried, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound G1-9. LC-MS (ESI): m / z = 648.2 [M+H] + 。

[0203] Step I

[0204] Compound G1-9 (1.04 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), N,N-diisopropylethylamine (0.6 g, 4.8 mmol) was added, the temperature was cooled to 0 - 10 °C, and trifluoromethanesulfonic anhydride (0.7 g, 2.4 mmol, 400 μL) was slowly added to the reaction solution. The reaction was carried out at this temperature for 15 min. The reaction solution was poured into saturated ammonium chloride aqueous solution (8 mL), separated, the aqueous phase was extracted with dichloromethane (5 mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound G1. LC-MS (ESI): m / z = 780.1 [M+H] + 。

[0205] Preparation of H1

[0206]

[0207] Step A

[0208] In a dry 1 L three-necked flask, potassium carbonate (27.6 g, 200 mmol) was added to N,N-dimethylformamide (100 mL), the temperature was cooled to 0 °C, and a solution of compound H1-1 (14.88 g, 80 mmol) in N,N-dimethylformamide (500 mL) was added dropwise under nitrogen protection. The reaction was continued at 0 °C for 0.5 h, p-methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20 °C, and the stirring was continued for 7.5 h under nitrogen protection. The reaction solution was slowly added to 100 mL of saturated ammonium chloride, extracted with methyl tert-butyl ether (100 mL × 2), the combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by silica gel column chromatography to obtain compound H1-2. LC-MS (ESI): m / z = 426.1 [M+H] + 。

[0209] Step B

[0210] 2,2,6,6 - Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL). The temperature was lowered to -5 °C, and n-butyllithium (2.5 M, 50 mL) was added dropwise. The reaction was carried out at -5 to 0 °C for 15 minutes. Then the temperature was lowered to -60 °C, and a solution of compound H1-2 (26.52 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The reaction was carried out at -60 °C for 0.5 hour. N,N-Dimethylformamide (22.8 g, 0.312 mol) was quickly added, and the reaction mixture was stirred at -60 °C for an additional 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction mixture, and it was extracted with methyl tert-butyl ether (100 mL × 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a solvent for 0.5 hour, filtered, and the filter cake was dried. The filtrate was concentrated and then purified by silica gel column chromatography. The filter cake and the column chromatography fractions were combined to obtain compound H1-3. LC-MS (ESI): m / z = 376.2 [M + H] + 。

[0211] Step D

[0212] Compound H1-3 (9.34 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and N-bromosuccinimide (4.5 g, 25 mmol) was added. The reaction mixture was stirred at 20 °C for an additional 20 minutes. The reaction mixture was added to 150 mL of water and extracted with methyl tert-butyl ether (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was slurried in a mixed solvent for 0.5 hour, filtered, and the filter cake was dried to obtain compound H1-4. LC-MS (ESI): m / z = 454.0 [M + H] + 。

[0213] Step E

[0214] Compound H1-4 (9.24 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Under nitrogen, cuprous iodide (7.8 g, 4.1 mmol) and methyl fluorosulfonyldifluoroacetate (20 g, 100 mmol) were added to the reaction mixture. The reaction mixture was heated to 100 °C and stirred for 1 hour. After cooling, the reaction mixture was filtered through diatomaceous earth. The filtrate was added to 150 mL of water and extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to obtain compound H1-5. LC-MS (ESI): m / z = 444.1 [M + H] + 。

[0215] Step E

[0216] Under nitrogen protection, H1-5 (5.76 g, 13 mmol) and lithium hydroxide (0.82 g, 19.5 mmol) were dissolved in 100 mL of THF at room temperature. Then dimethyl ethylphosphonoacetate (3.55 g, 19.5 mmol) was added, and the reaction mixture was stirred at room temperature for 24 hours. THF was recovered, and the mixture was added to 150 mL of water. It was extracted with methyl tert-butyl ether (75 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound H1-6. LC-MS (ESI): m / z = 514.1 [M+H] + 。

[0217] Step F

[0218] Compound H1-6 (10.26 g, 20 mmol) and piperidine (0.2 mL) were added to dichloromethane (100 mL). Ethyl 3-mercaptopropionate (3.2 g, 24 mmol) and benzyltrimethylammonium hydroxide (1 mL, 30% methanol) were added, and the mixture was stirred at 60 °C for 48 hours. Dilute hydrochloric acid was added, and it was extracted with dichloromethane (75 mL × 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound H1-7. LC-MS (ESI): m / z = 648.3 [M+H] + 。

[0219] Step G

[0220] Compound H1-7 (7.76 g, 12 mmol) and sodium ethoxide (1.36 g, 19 mmol) were dissolved in 400 mL of toluene and stirred at room temperature for 24 hours. It was poured into 100 mL of water and extracted with dichloromethane (100 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic solvents were removed under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography to obtain compound H1-8. LC-MS (ESI): m / z = 602.2 [M+H] + 。

[0221] Step H

[0222] To a suspension of H1-8 (3.19 g, 5.3 mmol) in water (10 mL), S-methylisothiourea sulfate (1.7 g, 6.2 mmol) and potassium carbonate (1.6 g, 11.7 mmol) were added at room temperature. The reaction mixture was stirred for 20 h. The precipitate was collected, washed with water (twice) and isopropyl ether, dried, and the crude product obtained was separated and purified by silica gel column chromatography to give compound H1-9. LC-MS (ESI): m / z = 628.2 [M+H] + 。

[0223] Step I

[0224] Compound H1-9 (1.00 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), N,N-diisopropylethylamine (0.6 g, 4.8 mmol) was added, the temperature was lowered to 0 - 10 °C, and trifluoromethanesulfonic anhydride (0.7 g, 2.4 mmol, 400 μL) was slowly added to the reaction solution. The reaction was carried out at this temperature for 15 min. The reaction solution was poured into saturated ammonium chloride aqueous solution (8 mL), liquid separation was carried out, the aqueous phase was extracted with dichloromethane (5 mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the organic solvent was removed under reduced pressure, and the crude product obtained was separated and purified by silica gel column chromatography to give compound H1. LC-MS (ESI): m / z = 760.1 [M+H] + 。

[0225] Example 1

[0226] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 1)

[0227]

[0228] Step A

[0229] Compound A1 (82 mg, 105.02 μmol) and 8-tert-butoxycarbonyl-3,8-diazabicyclo[3.2.1]octane (26.63 mg, 126.03 μmol) were dissolved in N,N-dimethylformamide (2 mL), diisopropylethylamine (40.72 mg, 315.07 μmol) was added, and the reaction solution was heated to 100 °C and stirred for an additional 1 h. After cooling, the organic solvent was removed under reduced pressure, and the crude product obtained was separated and purified by preparative thin layer chromatography plate to give compound 1-1. LC / MS (ESI): m / z = 840 [M+H] + 。

[0230] Step B

[0231] Dissolve compound 1-1 (71 mg, 84.96 μmol) in dichloromethane (2 mL), add m-chloroperbenzoic acid (34.50 mg, 169.92 μmol), and continue to stir the reaction solution at 20 °C for 3 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin-layer chromatography to obtain compound 1-2. LC / MS (ESI): m / z = 904 [M+H] + .

[0232] Step C

[0233] Under ice-water bath conditions, dissolve compound ((2R,7aS)-2-fluoro-1,2,3,7a-tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), continue to stir the reaction solution for 30 minutes, add a toluene (1 mL) solution of compound 1-2 (53 mg, 58.42 μmol), and continue to stir the reaction solution under ice-water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin-layer chromatography to obtain compound 1-3. LC / MS (ESI): m / z = 983.1 [M+H] + .

[0234] Step D

[0235] Dissolve compound 1-3 (42 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high performance liquid chromatography to obtain compound 1. LC / MS (ESI): m / z = 642.7 [M+H] + . 1 1H-NMR (DMSO-d6) δ 1.78 - 2.39 (m, 12H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 6.84 (d, 1H).

[0236] Example 2

[0237] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 1A)

[0238] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 1B)

[0239]

[0240] Compound 1 was separated and purified by supercritical liquid chromatography (SFC) to obtain A1 and A2 (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + )

[0241] Example 3

[0242] 4-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-7-(2-fluoro-3-hydroxy-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 2)

[0243]

[0244] Compound 2 - 4 was obtained by a preparation method similar to that of Compound 1 in Example 1 (using B1 as the raw material). LC / MS (ESI): m / z = 733.8 [M+H] +

[0245] ​Compound 2-4 (0.1 g, 1.4 mmol) was added to a 1:1 IPA:THF (25 mL) solution, stirred, and 10% Pd / C (0.1 g) was added to the stirred solution. The reaction mixture was stirred at room temperature for 16 h under hydrogen gas, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain Compound 2. LC / MS (ESI): m / z = 643.7 [M+H] + 。

[0246] Example 4

[0247] (7S)-4-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-7-(2-fluoro-3-hydroxy-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 2A)

[0248] (7R)-4-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-7-(2-fluoro-3-hydroxy-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 2B)

[0249]

[0250] Compounds 2A and 2B were obtained by a preparation method similar to that of Compounds 1A and 1B in Example 2 (using Compound 2 as the raw material). LC / MS (ESI): m / z = 643.7 [M+H] + 。

[0251] Example 5

[0252] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 3)

[0253]

[0254] Compound 3 was obtained by a preparation method similar to that of Compound 1 in Column 1 (with the raw material replaced by C1). LC / MS (ESI): m / z = 625.7 [M+H] +.1 1H-NMR (CDCl3) δ 1.78 - 2.20 (m, 7H), 2.01 - 2.20 (m, 6H), 2.25 - 2.60 (m, 6H), 3.00 - 3.20 (m, 3H), 3.30 - 3.50 (m, 1H), 3.60 - 3.80 (m, 2H), 3.80 - 4.15 (m, 4H), 4.30 - 4.60 (m, 6H), 4.80 - 5.00 (m, 1H), 5.29 - 5.54 (m, 1H), 7.30 (s, 1H).

[0255] Example 6

[0256] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 3A)

[0257] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 3B)

[0258]

[0259] Compound 3A and 3B were obtained by a preparation method similar to that of Compounds 1A and 1B in Example 2 (with the raw material replaced by Compound 3). LC / MS (ESI): m / z = 625.7 [M+H] + 。

[0260] Example 7

[0261] 4-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7-(6-hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 4)

[0262]

[0263] Compound 4 was obtained by a preparation method similar to that of Compound 2 in Example 3 (using D1 as the raw material). LC / MS (ESI): m / z = 626.7 [M+H] + 。

[0264] Example 8

[0265] (7S)-4-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7-(6-hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 4A)

[0266] (7R)-4-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7-(6-hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 4B)

[0267]

[0268] Compound 4A and 4B were obtained by a preparation method similar to that of Compound 1A and 1B in Example 2 (using Compound 4 as the raw material). LC / MS (ESI): m / z = 626.7 [M+H] + 。

[0269] Example 9

[0270] 7-(3-Amino-2-fluoro-5-methyl-`6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 5)

[0271]

[0272] Compound 5 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 645.7 [M+H] + 。

[0273] Example 10

[0274] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 5A)

[0275] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 5B)

[0276]

[0277] Compound 5A and 5B were obtained by a preparation method similar to that of Compounds 1A and 1B in Example 2 (using Compound 5 as the raw material). LC / MS (ESI): m / z = 645.7 [M+H] + 。

[0278] Example 11

[0279] 7-(2-Fluoro-3-hydroxy-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 6)

[0280]

[0281] Compound 6 was obtained by a preparation method similar to that of Compound 2 in Example 3 (using B1 as the raw material). LC / MS (ESI): m / z = 646.7 [M+H] + 。

[0282] Example 12

[0283] (7S)-7-(2-Fluoro-3-hydroxy-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 6A)

[0284] (7R)-7-(2-Fluoro-3-hydroxy-5-methyl-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 6B)

[0285]

[0286] Compound 6A and Compound 6B were obtained by a preparation method similar to that of Compound 1A and Compound 1B in Example 2 (using Compound 6 as the raw material). LC / MS (ESI): m / z = 646.7 [M+H] + 。

[0287] Example 13

[0288] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 7)

[0289]

[0290] Compound 7 was obtained by a preparation method similar to that of Compound 1 in Example 1 (using C1 as the raw material). LC / MS (ESI): m / z = 628.7 [M+H] + 。

[0291] Example 14

[0292] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 7A)

[0293] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 7B)

[0294]

[0295] Compounds 7A and 7B were obtained by a preparation method similar to that of Compounds 1A and 1B in Example 2 (using Compound 7 as the raw material). LC / MS (ESI): m / z = 628.7 [M+H] + 。

[0296] Example 15

[0297] 2-(((2R,7aS)-2-Fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7-(6-hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 8)

[0298]

[0299] Compound 8 was obtained by a preparation method similar to that of Compound 2 in Example 3 (using D1 as the raw material). LC / MS (ESI): m / z = 629.7 [M+H] + 。

[0300] Example 16

[0301] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 8A)

[0302] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 8B)

[0303]

[0304] Compounds 8A and 8B were obtained by a preparation method similar to that of Compounds 1A and 1B in Example 2 (using Compound 8 as the raw material). LC / MS (ESI): m / z = 629.7 [M+H] + 。

[0305] Example 17

[0306] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 9)

[0307]

[0308] Step A

[0309] Under ice-water bath conditions, dissolve ((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), continue to stir the reaction solution for 30 minutes, add a toluene (1 mL) solution of Compound 1-2 (53 mg, 58.42 μmol), and continue to stir the reaction solution under ice-water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin-layer chromatography plate to obtain Compound 1-3. LC / MS (ESI): m / z = 983.1 [M+H] + 。

[0310] Step B

[0311] Dissolve Compound 1-3 (42 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high performance liquid chromatography to obtain Compound 17. LC / MS (ESI): m / z = 642.7 [M+H] + 。 1 1H-NMR (DMSO-d6) δ 1.99 (m, 4H), 2.10 - 2.27 (m, 3H), 2.84 (d, 1H), 2.98 (d, 1H), 3.29 - 3.64 (m, 3H), 3.84 (d, 1H), 4.09 - 4.33 - 4.76 (m, 13H), 5.96 (s, 2H) 6.84 (d, 1H).

[0312] Example 18

[0313] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 9A)

[0314] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 9B)

[0315]

[0316] Compound 9 was separated and purified by supercritical liquid chromatography (SFC) to obtain 9A and 9B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + )

[0317] Example 19

[0318] 7-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 10)

[0319]

[0320] Step A

[0321] Under ice - water bath conditions, dissolve the compound ((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.98 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert - butoxide (7.30 mg, 75.94 μmol), stir the reaction solution for another 30 minutes, add a toluene (1 mL) solution of compound 1 - 2 (53 mg, 58.42 μmol), and continue to stir the reaction solution under ice - water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin - layer chromatography plate to obtain compound 10 - 1. LC / MS(ESI): m / z = 995.4[M + H] + 。

[0322] Step B

[0323] Dissolve compound 10 - 1 (43 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high - performance liquid chromatography to obtain compound 10. LC / MS(ESI): m / z = 655.2[M + H] + 。 1 1H - NMR(DMSO - d6) δ 1.68 - 2.31(m, 12H), 2.75(d, 1H), 2.94(d, 1H), 3.17 - 3.61(m, 5H), 4.05 - 4.62(m, 11H), 5.20(d, 2H), 5.96(s, 2H), 6.84 - 7.05(m, 3H).

[0324] Example 20

[0325] (7S)-7-(3 - Amino - 2 - fluoro - 5 - methyl - 6-(trifluoromethyl)phenyl)-4-(3,8 - diazabicyclo[3.2.1]octan - 3 - yl)-2-(((S)-2-(difluoromethylene)tetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-7,8 - dihydro - 5H - thiopyrano[4,3 - d]pyrimidine - 6,6 - dioxide (Compound 10A)

[0326] (7R)-7-(3 - Amino - 2 - fluoro - 5 - methyl - 6-(trifluoromethyl)phenyl)-4-(3,8 - diazabicyclo[3.2.1]octan - 3 - yl)-2-(((S)-2-(difluoromethylene)tetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-7,8 - dihydro - 5H - thiopyrano[4,3 - d]pyrimidine - 6,6 - dioxide (Compound 10B)

[0327]

[0328] Compound 10 was separated and purified by supercritical liquid chromatography (SFC) to obtain 10A and 10B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 655.2 [M+H] + )

[0329] Example 21

[0330] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 11)

[0331]

[0332] Step A

[0333] Under ice-water bath conditions, ((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (11.62 mg, 75.94 μmol) was dissolved in anhydrous toluene (1 mL), sodium tert-butoxide (7.30 mg, 75.94 μmol) was added, the reaction solution was stirred for another 30 minutes, a toluene (1 mL) solution of Compound 1-2 (53 mg, 58.42 μmol) was added, and the reaction solution was stirred under ice-water bath for 2 hours. The organic solvent was removed under reduced pressure, and the obtained crude product was separated and purified by preparative thin-layer chromatography plate to obtain Compound 11-1. LC / MS (ESI): m / z = 977.1 [M+H] + 。

[0334] Step B

[0335] Compound 11-1 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the reaction solution was stirred at 20 °C for 2 hours. The solvent was removed under reduced pressure, and the obtained crude product was separated and purified by high performance liquid chromatography to obtain Compound 11. LC / MS (ESI): m / z = 637.3 [M+H] + 。 11H-NMR (DMSO-d6) δ 1.79–2.31 (m, 12H), 2.55 - 2.75 (m, 1H), 2.94 (d, 1H), 3.17 - 3.61 (m, 5H), 4.05 - 4.62 (m, 11H), 5.20 (d, 2H), 6.84 (d, 1H).

[0336] Example 22

[0337] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 11A)

[0338] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 11B)

[0339]

[0340] Compound 11 was separated and purified by supercritical liquid chromatography (SFC) to obtain 11A and 11B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 - 50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + )

[0341] Example 23

[0342] 7-(6-Hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 12)

[0343]

[0344] Compound 12 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 640.3 [M+H] + 。

[0345] Example 24

[0346] (7S)-7-(6-Hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 12A)

[0347] [[ID=]](7R)-7-(6-Hydroxy-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 12B)

[0348]

[0349] Compound 12A and 12B were obtained by a preparation method similar to that of Compound 2A and 2B in Example 2 (using Compound 12 as the raw material). LC / MS (ESI): m / z == 640.3 [M+H] + 。

[0350] Example 25

[0351] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 13)

[0352]

[0353] Compound 13 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 658.2 [M+H] + 。

[0354] Example 26

[0355] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 13A)

[0356] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 13B)

[0357]

[0358] Compound 13A and 13B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 13 as the raw material). LC / MS (ESI): m / z = 658.2 [M+H] + 。

[0359] Example 27

[0360] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 14)

[0361]

[0362] Compound 14 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 676.2 [M+H] + 。

[0363] Example 28

[0364] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 14A)

[0365] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 14B)

[0366]

[0367] Compounds 14A and 14B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 14 as the raw material). LC / MS (ESI): m / z = 676.2 [M+H] + 。

[0368] Example 29

[0369] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 15)

[0370]

[0371] Compound 15 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 659.2 [M+H] + 。

[0372] Example 30

[0373] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 15A)

[0374] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 15B)

[0375]

[0376] Compounds 15A and 15B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 15 as the raw material). LC / MS (ESI): m / z 659.2 [M+H] + 。

[0377] Example 31

[0378] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 16)

[0379]

[0380] Compound 16 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 641.2 [M+H] + 。

[0381] Example 32

[0382] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 16A)

[0383] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 16B)

[0384]

[0385] Compounds 16A and 16B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 16 as the raw material). LC / MS (ESI): m / z = 641.2 [M+H] + 。

[0386] Example 33

[0387] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 17)

[0388]

[0389] Compound 17 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 622.3 [M+H] + 。

[0390] Example 34

[0391] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 17A)

[0392] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 17B)

[0393]

[0394] Compound 17A and 17B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 17 as the raw material). LC / MS (ESI): m / z = 622.3 [M+H] + 。

[0395] Example 35

[0396] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(difluoromethylidene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 18)

[0397]

[0398] Step A

[0399] Under ice - water bath conditions, dissolve the compound ((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (14.28 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), stir the reaction solution for another 30 minutes, add a toluene (1 mL) solution of compound 1-2 (53 mg, 58.42 μmol), and continue to stir the reaction solution under ice - water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin - layer chromatography plate to obtain compound 18-1. LC / MS(ESI): m / z = 996.1 [M + H] + 。

[0400] Step B

[0401] Dissolve compound 18-1 (43 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high - performance liquid chromatography to obtain compound 18. LC / MS(ESI): m / z = 656.2 [M + H] + 。 1 1H - NMR(CDCl3) δ 1.79–2.31(m, 12H), 2.75(d, 1H), 2.94(d, 1H), 3.17 - 3.61(m, 5H), 4.05 - 4.68(m, 11H), 5.20(d, 2H), 7.30(s, 1H).

[0402] Example 36

[0403] (7S)-7-(6 - Amino - 4 - methyl - 3-(trifluoromethyl)pyridin - 2 - yl)-2-(((S)-2-(difluoromethylene)tetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-4-(3,8 - diazabicyclo[3.2.1]octan - 3 - yl)-7,8 - dihydro - 5H - thiopyrano[4,3 - d]pyrimidine - 6,6 - dioxide (Compound 18A)

[0404] (7R)-7-(6 - Amino - 4 - methyl - 3-(trifluoromethyl)pyridin - 2 - yl)-2-(((S)-2-(difluoromethylene)tetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-4-(3,8 - diazabicyclo[3.2.1]octan - 3 - yl)-7,8 - dihydro - 5H - thiopyrano[4,3 - d]pyrimidine - 6,6 - dioxide (Compound 18B)

[0405]

[0406] Compound 18 was separated and purified by supercritical liquid chromatography (SFC) to obtain 18A and 18B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 656.2 [M+H] + )

[0407] Example 37

[0408] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 19)

[0409]

[0410] Step A

[0411] Under ice-water bath conditions, ((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.98 mg, 75.94 μmol) was dissolved in anhydrous toluene (1 mL), sodium tert-butoxide (7.30 mg, 75.94 μmol) was added, the reaction solution was stirred for an additional 30 minutes, a toluene (1 mL) solution of Compound 1-2 (53 mg, 58.42 μmol) was added, and the reaction solution was stirred under ice-water bath for 2 hours. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by a preparative thin-layer chromatography plate to obtain Compound 1-3. LC / MS (ESI): m / z = 983.1 [M+H] +

[0412] Step B

[0413] Compound 1-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the reaction solution was stirred at 20 °C for 2 hours. The solvent was removed under reduced pressure, and the resulting crude product was separated and purified by high-performance liquid chromatography to obtain Compound 1. LC / MS (ESI): m / z = 638.3 [M+H] +

[0414] Example 38​​

[0415] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 19A)

[0416] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 19B)

[0417]

[0418] Compound 19 was separated and purified by supercritical liquid chromatography (SFC) to obtain 19A and 19B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 638.3 [M+H] + )

[0419] Example 39

[0420] 7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 20)

[0421]

[0422] Step A

[0423] Under ice-water bath conditions, dissolve the compound ((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (11.63 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), continue to stir the reaction solution for 30 minutes, add a toluene (1 mL) solution of compound 1-2 (53 mg, 58.42 μmol), and continue to stir the reaction solution under ice-water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin-layer chromatography plate to obtain compound 20-1. LC / MS (ESI): m / z = 983.1 [M+H] + 。

[0424] Step B

[0425] Dissolve compound 20-1 (42 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high performance liquid chromatography to obtain compound 1. LC / MS (ESI): m / z = 620.3 [M+H] + 。 1 1H-NMR (CDCl3) δ 1.78–2.31 (m, 12H), 2.55-75 (m, 1H), 2.94 (d, 1H), 3.17-3.61 (m, 5H), 4.Oh-4.62 (m, 11H), 5.20 (d, 2H), 7.30 (s, 1H).

[0426] Example 40

[0427] (7S)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 20A)

[0428] (7R)-7-(6-Amino-4-methyl-3-(trifluoromethyl)pyridin-2-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 20B)

[0429]

[0430] Compound 20 was separated and purified by supercritical liquid chromatography (SFC) to obtain 20A and 20B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 620.3 [M+H] + )

[0431] Example 41

[0432] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 21)

[0433]

[0434] Step A

[0435] Compound E1 (84 mg, 105.02 μmol) and 8-tert-butoxycarbonyl-3,8-diazabicyclo[3.2.1]octane (26.63 mg, 126.03 μmol) were dissolved in N,N-dimethylformamide (2 mL), diisopropylethylamine (40.72 mg, 315.07 μmol) was added, and the reaction mixture was heated to 100 °C and stirred for an additional 1 hour. After cooling, the organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain Compound 21-1. LC / MS (ESI): m / z = 864 [M+H] + 。

[0436] Step B

[0437] Compound 21-1 (73 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), m-chloroperoxybenzoic acid (34.50 mg, 169.92 μmol) was added, and the reaction mixture was stirred at 20 °C for an additional 3 hours. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain Compound 21-2. LC / MS (ESI): m / z = 928 [M+H] + 。

[0438] Step C

[0439] Under ice-water bath conditions, dissolve the compound ((2R,7aS)-2-fluoro-tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), continue to stir the reaction solution for 30 minutes, add a toluene (1 mL) solution of compound 21-2 (60 mg, 58.42 μmol), and continue to stir the reaction solution under ice-water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin-layer chromatography plate to obtain compound 21-3. LC / MS (ESI): m / z = 1007.4 [M+H] + 。

[0440] Step D

[0441] Dissolve compound 21-3 (43 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high performance liquid chromatography to obtain compound 21. LC / MS (ESI): m / z = 667.2 [M+H] + 。 1 1H-NMR (CDCl3) δ 1.78 - 2.20 (m, 7H), 2.01 - 2.20 (m, 6H), 2.25 - 2.60 (m, 6H), 3.00 - 3.20 (m, 2H), 3.30 - 3.50 (m, 1H), 3.60 - 3.80 (m, 2H), 3.80 - 4.15 (m, 4H), 4.30 - 4.60 (m, 6H), 4.80 - 5.00 (m, 1H), 5.29 - 5.54 (m, 1H), 6.82 (d, 1H).

[0442] Example 42

[0443] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluoro-tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 21A)

[0444] (7R)-7-(3-amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7AS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 21B)

[0445]

[0446] Compound 21 was separated and purified by supercritical fluid chromatography (SFC) to give 21A and 21B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 667.2 [M+H] + ).

[0447] Example 43

[0448] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 22)

[0449]

[0450] Step A

[0451] In an ice-water bath, compound ((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (11.63 mg, 75.94 μmol) was dissolved in anhydrous toluene (1 mL). Sodium tert-butoxide (7.30 mg, 75.94 μmol) was added, and the reaction solution was stirred for 30 minutes. A solution of compound 1-2 (53 mg, 58.42 μmol) in toluene (1 mL) was added, and the reaction solution was stirred for 2 hours in an ice-water bath. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain compound 22-1. LC / MS (ESI): m / z = 1001.4 [M+H] + .

[0452] Step D

[0453] Compound 22-1 (43 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the reaction mixture was stirred at 20 °C for an additional 2 hours. The solvent was removed under reduced pressure, and the resulting crude product was separated and purified by high performance liquid chromatography to obtain Compound 1. LC / MS (ESI): m / z = 661.2 [M+H] + 。 1 1H-NMR (CDCl3) δ 1.79–2.31 (m, 12H), 2.75 (d, 1H), 2.94 (d, 1H), 3.17 - 3.61 (m, 5H), 4.05 - 4.62 (m, 11H), 5.20 (d, 2H), 6.82 (d, 1H).

[0454] Example 44

[0455] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 22A)

[0456] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(methylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 22B)

[0457]

[0458] Compound 22 was separated and purified by supercritical fluid chromatography (SFC) to obtain 22A and 22B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 661.2 [M+H] + )。

[0459] Example 45

[0460] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 23)

[0461]

[0462] Step A

[0463] Under ice-water bath conditions, dissolve ((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (14.28 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), continue to stir the reaction solution for 30 minutes, add a toluene (1 mL) solution of Compound 1-2 (53 mg, 58.42 μmol), and continue to stir the reaction solution under ice-water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the obtained crude product by preparative thin-layer chromatography plate to obtain Compound 23-1. LC / MS (ESI): m / z = 1036.4 [M+H] + 。

[0464] Step B

[0465] Dissolve Compound 23-1 (44 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction solution at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the obtained crude product by high performance liquid chromatography to obtain Compound 23. LC / MS (ESI): m / z = 697.2 [M+H] + 。 1 1H-NMR (CDCl3) δ 1.73 - 2.03 (m, 11H), 2.21 - 2.33 (m, 1H), 2.36 - 2.46 (m, 1H), 2.74 (m, 1H), 2.80 (d, 1H), 3.13 - 3.35 (m, 5H), 3.66 (m, 2H), 4.21 - 4.46 (m, 9H), 6.82 (d, 1H).

[0466] Example 46

[0467] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 23A)

[0468] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 23B)

[0469]

[0470] Compound 23 was separated and purified by supercritical fluid chromatography (SFC) to obtain 23A and 23B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 697.2 [M+H] + )

[0471] Example 47

[0472] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 24)

[0473]

[0474] Step A

[0475] Under ice-water bath conditions, dissolve the compound ((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.98 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), stir the reaction mixture for an additional 30 minutes, add a toluene (1 mL) solution of compound 1-2 (53 mg, 58.42 μmol), and continue to stir the reaction mixture under ice-water bath for 2 hours. Remove the organic solvent under reduced pressure, and purify the resulting crude product by preparative thin-layer chromatography to obtain compound 1-3. LC / MS (ESI): m / z = 1019.1 [M+H] + 。

[0476] Step D

[0477] Dissolve compound 1-3 (43 mg, 42.78 μmol) in anhydrous dichloromethane (2 mL), add trifluoroacetic acid (1 mL), and continue to stir the reaction mixture at 20 °C for 2 hours. Remove the solvent under reduced pressure, and purify the resulting crude product by high performance liquid chromatography to obtain compound 1. LC / MS (ESI): m / z = 679.2 [M+H] + 。 1 1H-NMR (CDCl3) δ 1.71 - 2.20 (m, 5H), 2.01 - 2.20 (m, 6H), 2.25 - 2.60 (m, 6H), 3.00 - 3.20 (m, 3H), 3.30 - 3.50 (m, 1H), 3.60 - 3.80 (m, 2H), 3.80 - 4.15 (m, 4H), 4.30 - 4.60 (m, 6H), 4.80 - 5.00 (m, 1H), 5.29 - 5.54 (m, 1H), 6.46 - 6.71 (m, 1H), 6.84 (d, 1H).

[0478] Example 48

[0479] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 24A)

[0480] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 24B)

[0481]

[0482] Compound 24 was separated and purified by supercritical liquid chromatography (SFC) to obtain 24A and 24B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 679.2 [M+H] + )

[0483] Example 49

[0484] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 25)

[0485]

[0486] Compound 25 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 664.2 [M+H] + 。

[0487] Example 50

[0488] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 25A)

[0489] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 25B)

[0490]

[0491] Compound 25A and 25B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 25 as the raw material). LC / MS (ESI): m / z = 664.2 [M+H] + 。

[0492] Example 51

[0493] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 26)

[0494]

[0495] Compound 5 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 682.2 [M+H] + 。

[0496] Example 52

[0497] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 26A)

[0498] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 26B)

[0499]

[0500] Compound 26A and 26B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 26 as the starting material). LC / MS (ESI): m / z = 682.2 [M+H] + 。

[0501] Example 53

[0502] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 27)

[0503]

[0504] Compound 5 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 700.2 [M+H] + 。

[0505] Example 54

[0506] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 27A)

[0507] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-2-(((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 27B)

[0508]

[0509] Compound 27A and 27B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 27 as the raw material). LC / MS (ESI): m / z = 700.2 [M+H] + 。

[0510] Example 55

[0511] 7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 28)

[0512]

[0513] Compound 28 was obtained by a preparation method similar to that of Compound 1 in Example 1. LC / MS (ESI): m / z = 670.2 [M+H] + 。

[0514] Example 56

[0515] (7S)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 28A)

[0516] (7R)-7-(3-Amino-2-fluoro-5-(prop-1-yn-1-yl)-6-(trifluoromethyl)phenyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 28B)

[0517]

[0518] Compounds 28A and 28B were obtained by a preparation method similar to that of Compounds 2A and 2B in Example 2 (using Compound 28 as the raw material). LC / MS (ESI): m / z = 670.2 [M+H] + 。

[0519] Example 57

[0520] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 29)

[0521]

[0522] Step A

[0523] Dissolve Compound F1 (82 mg, 105.02 μmol) and 8-tert-butoxycarbonyl-3,8-diazabicyclo[3.2.1]octane (26.63 mg, 126.03 μmol) in N,N-dimethylformamide (2 mL), add diisopropylethylamine (40.72 mg, 315.07 μmol), and heat the reaction mixture to 100 °C and continue stirring for 1 hour. Cool, remove the organic solvent under reduced pressure, and purify the resulting crude product by preparative thin-layer chromatography plate to obtain Compound 29-1. LC / MS (ESI): m / z = 846 [M+H] + 。

[0524] Step B

[0525] Compound 29-1 (71 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperoxybenzoic acid (34.50 mg, 169.92 μmol) was added. The reaction mixture was stirred at 20 °C for an additional 3 hours. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain Compound 29-2. LC / MS (ESI): m / z = 910 [M+H] + 。

[0526] Step C

[0527] Under an ice-water bath condition, ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 mg, 75.94 μmol) was dissolved in anhydrous toluene (1 mL), and sodium tert-butoxide (7.30 mg, 75.94 μmol) was added. The reaction mixture was stirred for an additional 30 minutes, and a toluene (1 mL) solution of Compound 29-2 (53 mg, 58.42 μmol) was added. The reaction mixture was stirred at an ice-water bath for an additional 2 hours. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain Compound 29-3. LC / MS (ESI): m / z = 989 [M+H] + 。

[0528] Step D

[0529] Compound 29-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 20 °C for an additional 2 hours. The solvent was removed under reduced pressure, and the resulting crude product was separated and purified by high performance liquid chromatography to obtain Compound 29. LC / MS (ESI): m / z = 648.2 [M+H] + 。 1 1H-NMR (DMSO-d6) δ 1.78 - 2.39 (m, 12H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 5.92 (s, 2H), 6.69 (d, 1H), 6.88 (d, 1H).

[0530] Example 58

[0531] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 29A)

[0532] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 29B)

[0533]

[0534] Compound 29 was separated and purified by supercritical liquid chromatography (SFC) to obtain 29A and 29A (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 648.2 [M+H] + )

[0535] Example 59

[0536] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 30)

[0537]

[0538] Compound 30 was obtained by a preparation method similar to that of Compound 29 in Example 57. LC / MS (ESI): m / z = 642.3 [M+H] + 。

[0539] Example 60

[0540] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 30A)

[0541] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 30B)

[0542]

[0543] Compound 30 was separated and purified by supercritical liquid chromatography (SFC) to obtain 30A and 30B (Column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; Mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); Gradient: B% = 5 to -50%, 3 min; Flow rate: 3.4 mL / min; Wavelength: 220 nm; Pressure: 1800, LC / MS (ESI): m / z = 642.3 [M+H] + )

[0544] Example 61

[0545] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 31)

[0546]

[0547] Compound 31 was obtained by a preparation method similar to that of Compound 29 in Example 57. LC / MS (ESI): m / z = 679.2 [M+H] + 。 1H-NMR (DMSO-d6) δ 1.99 (m, 4H), 2.10 - 2.27 (m, 3H), 2.84 (d, 1H), 2.98 (d, 1H), 3.29 - 3.64 (m, 3H), 3.84 (d, 1H), 4.09 - 4.33 - 4.76 (m, 13H), 5.92 (s, 2H), 6.69 (d, 1H), 6.88 (d, 1H).

[0548] Example 62

[0549] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 31A)

[0550] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 31B)

[0551]

[0552] Compound 31 was separated and purified by supercritical liquid chromatography (SFC) to obtain 31A and 31B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 - -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 679.2 [M + H] + ).

[0553] Example 63

[0554] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 32)

[0555]

[0556] Compound 32 was obtained by a preparation method similar to that of Compound 29 in Example 57. LC / MS (ESI): m / z = 661.2 [M+H] + 。 1 1H-NMR (DMSO-d6) δ 1.78 - 2.39 (m, 10H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 5.92 (s, 2H), 6.59 - 6.87 (m, 3H).

[0557] Example 64

[0558] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 32A)

[0559] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 32B)

[0560]

[0561] Compound 32 was separated and purified by supercritical liquid chromatography (SFC) to obtain 32A and 32B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 - -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 661.2 [M+H] + )。

[0562] Example 65

[0563] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 33)

[0564]

[0565] Step A

[0566] Dissolve Compound G1 (82 mg, 105.02 μmol) and 8-tert-butoxycarbonyl-3,8-diazabicyclo[3.2.1]octane (26.63 mg, 126.03 μmol) in N,N-dimethylformamide (2 mL), add diisopropylethylamine (40.72 mg, 315.07 μmol), heat the reaction mixture to 100 °C and stir for an additional 1 hour. Cool, remove the organic solvent under reduced pressure, and purify the resulting crude product by preparative thin-layer chromatography to obtain Compound 33-1. LC / MS (ESI): m / z = 842 [M+H] + 。

[0567] Step B

[0568] Dissolve Compound 33-1 (71 mg, 84.96 μmol) in dichloromethane (2 mL), add m-chloroperoxybenzoic acid (34.50 mg, 169.92 μmol), and stir the reaction mixture at 20 °C for an additional 3 hours. Remove the organic solvent under reduced pressure, and purify the resulting crude product by preparative thin-layer chromatography to obtain Compound 33-2. LC / MS (ESI): m / z = 906 [M+H] + 。

[0569] Step C

[0570] Under ice-water bath conditions, dissolve ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 mg, 75.94 μmol) in anhydrous toluene (1 mL), add sodium tert-butoxide (7.30 mg, 75.94 μmol), stir the reaction mixture for 30 minutes, add a toluene (1 mL) solution of Compound 33-2 (53 mg, 58.42 μmol), and stir the reaction mixture under ice-water bath for an additional 2 hours. Remove the organic solvent under reduced pressure, and purify the resulting crude product by preparative thin-layer chromatography to obtain Compound 33-3. LC / MS (ESI): m / z = 985.1 [M+H] + 。

[0571] Step D

[0572] Compound 33-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the reaction mixture was stirred at 20 °C for an additional 2 hours. The solvent was removed under reduced pressure, and the resulting crude product was separated and purified by high performance liquid chromatography to obtain Compound 1. LC / MS (ESI): m / z = 645.2 [M+H] + . 1 1H-NMR (DMSO-d6) δ 1.78 - 2.39 (m, 12H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 6.20 (s, 2H), 6.69 (d, 1H), 6.89 (d, 1H).

[0573] Example 66

[0574] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 33A)

[0575] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 33B)

[0576]

[0577] Compound 33 was separated and purified by supercritical fluid chromatography (SFC) to obtain 33A and 33B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + ).

[0578] Example 67

[0579] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 34)

[0580]

[0581] Compound 34 was obtained by a preparation method similar to that of Compound 33 in Example 65. LC / MS (ESI): m / z = 638.2 [M+H] + 。

[0582] Example 68

[0583] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 34A)

[0584] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 34B)

[0585]

[0586] Compound 34 was separated and purified by supercritical liquid chromatography (SFC) to obtain 34A and 34B (column: Chiralpak IG-3: 3μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 638.2 [M+H] + )。

[0587] Example 69

[0588] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 35)

[0589]

[0590] Compound 35 was obtained by a preparation method similar to that of Compound 33 in Example 65. LC / MS (ESI): m / z = 637.2 [M+H] + 。 1 1H-NMR (DMSO-d6) δ 1.67 - 2.39 (m, 10H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 5.78 (s, 2H), 6.43 (s, 1H), 6.74 (s, 1H), 6.76 - 7.19 (m, 1H).

[0591] Example 70

[0592] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 35A)

[0593] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 35B)

[0594]

[0595] Compound 35 was separated and purified by supercritical liquid chromatography (SFC) to obtain 35A and 35B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + )

[0596] Example 71

[0597] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 36)

[0598]

[0599] Compound 36 was obtained by a preparation method similar to that of Compound 33 in Example 65. LC / MS (ESI): m / z = 619.2 [M+H] + 。 1 1H-NMR (DMSO-d6) δ 1.60 - 2.39 (m, 9H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 6.31 (s, 2H), 6.74 (d, 1H), 6.92 (d, 1H).

[0600] Example 72

[0601] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 36A)

[0602] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 36B)

[0603]

[0604] Compound 36 was separated and purified by supercritical liquid chromatography (SFC) to obtain 36A and 36B (column: Chiralpak IG-3: 3μm 0.46cm×5cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5~-50%, 3min; flow rate: 3.4mL / min; wavelength: 220nm; pressure: 1800, LC / MS (ESI): m / z = 619.2 [M+H] + )

[0605] Example 73

[0606] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 37)

[0607]

[0608] Step A

[0609] Compound H1 (80mg, 105.02μmol) and 8-tert-butoxycarbonyl-3,8-diazabicyclo[3.2.1]octane (26.63mg, 126.03μmol) were dissolved in N,N-dimethylformamide (2mL), diisopropylethylamine (40.72mg, 315.07μmol) was added, and the reaction solution was heated to 100°C and stirred for 1 hour. After cooling, the organic solvent was removed under reduced pressure, and the obtained crude product was separated and purified by preparative thin-layer chromatography plate to obtain Compound 37-1. LC / MS (ESI): m / z = 822 [M+H] +

[0610] Step B

[0611] ​Compound 37-1 (70 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperoxybenzoic acid (34.50 mg, 169.92 μmol) was added. The reaction mixture was stirred at 20 °C for 3 hours. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain compound 37-2. LC / MS (ESI): m / z = 886 [M+H] + 。

[0612] Step C

[0613] Under an ice-water bath condition, ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 mg, 75.94 μmol) was dissolved in anhydrous toluene (1 mL), and sodium tert-butoxide (7.30 mg, 75.94 μmol) was added. The reaction mixture was stirred for 30 minutes, and then a toluene (1 mL) solution of compound 37-2 (52 mg, 58.42 μmol) was added. The reaction mixture was stirred at an ice-water bath for 2 hours. The organic solvent was removed under reduced pressure, and the resulting crude product was separated and purified by preparative thin-layer chromatography to obtain compound 37-3. LC / MS (ESI): m / z = 965.1 [M+H] + 。

[0614] Step D

[0615] Compound 37-2 (41 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 20 °C for 2 hours. The solvent was removed under reduced pressure, and the resulting crude product was separated and purified by high-performance liquid chromatography to obtain compound 37. LC / MS (ESI): m / z = 625.3 [M+H] + 。 1 1H-NMR (DMSO-d6) δ 1.78 - 2.39 (m, 12H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 5.77 (s, 2H), 6.41 (s, 1H), 6.74 (s, 1H).

[0616] Example 74

[0617] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 37A)

[0618] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 37B)

[0619]

[0620] Compound 37 was separated and purified by supercritical liquid chromatography (SFC) to obtain 37A and 37B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + )

[0621] Example 75

[0622] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 38)

[0623]

[0624] Compound 38 was obtained by a preparation method similar to that of Compound 37 in Example 73. LC / MS (ESI): m / z = 619.2 [M+H] +

[0625] Example 76

[0626] ​(7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 38A)

[0627] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 38B)

[0628]

[0629] Compound 38 was separated and purified by supercritical liquid chromatography (SFC) to obtain 38A and 39B (Column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; Mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); Gradient: B% = 5 to -50%, 3 min; Flow rate: 3.4 mL / min; Wavelength: 220 nm; Pressure: 1800, LC / MS (ESI): m / z = 619.2 [M+H] + )

[0630] Example 77

[0631] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 39)

[0632]

[0633] Compound 39 was obtained by a preparation method similar to that of Compound 37 in Example 73. LC / MS (ESI): m / z = 637.2 [M+H] + 。 11H-NMR (DMSO-d6) δ 1.60 - 2.39 (m, 10H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 5.76 (s, 2H), 6.41 (s, 1H), 6.58 - 6.88 (m, 2H).

[0634] Example 78

[0635] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 39A)

[0636] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 39B)

[0637]

[0638] Compound 39 was separated and purified by supercritical liquid chromatography (SFC) to obtain 39A and 39B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 - 50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 637.2 [M + H] + )

[0639] Example 79

[0640] 7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-(1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 40)

[0641]

[0642] Compound 40 was obtained by a preparation method similar to that of Compound 37 in Example 73. LC / MS (ESI): m / z == 655.2 [M+H] + 。 1 H-NMR (DMSO-d6) δ 1.66 - 2.39 (m, 10H), 2.38 - 2.79 (m, 3H), 3.13 - 3.44 (m, 3H), 3.56 - 3.62 (m, 2H), 3.71 - 4.43 (m, 10H), 5.46 - 5.58 (m, 1H), 5.76 (s, 2H), 6.41 (s, 1H), 6.74 (s, Example 80

[0643] (7S)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 40A)

[0644] (7R)-7-(3-Amino-2-fluoro-5-methyl-6-(trifluoromethyl)phenyl)-4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine 6,6-dioxide (Compound 40B)

[0645]

[0646] Compound 40 was separated and purified by supercritical liquid chromatography (SFC) to obtain 40A and 40B (column: Chiralpak IG-3: 3 μm, 0.46 cm × 5 cm; mobile phase: A (CO2) and B (EtOH, containing 0.1% isopropylamine); gradient: B% = 5 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 655.2 [M+H] + )。

[0647] Example 81 Biological Activity Test

[0648] Experimental Example 1. KRAS Inhibitory Activity Test

[0649] 1. Experimental Purpose:

[0650] By the method of TR-FRET, compounds that can effectively inhibit KRas G12D from binding to GTP were screened out.

[0651] 2. Reagent preparation:

[0652] a. Storage reagents:

[0653] 1) KRAS nucleotide exchange buffer

[0654] Take 20 mL of 1000 mM HEPES, 20 mL of 500 mM EDTA, 10 mL of 5 M sodium chloride, 0.1 mL of 100% Tween 20, and 949.9 mL of water, prepare to make 1 L of solution, sterilize by filtration method, and store at 4 °C.

[0655] 2) KRAS experimental buffer

[0656] Take 20 mL of 1000 mM HEPES, 10 mL of 1000 mM magnesium chloride, 30 mL of 5 M sodium chloride, 0.05 mL of 100% Tween 20, and 939.95 mL of water, prepare to make 1 L of solution, sterilize by filtration method, and store at 4 °C.

[0657] 3) KRAS / Bodipy GDP / Tb-SA mixture

[0658] Take 9.5 μL of 95 μM KRas G12D protein, mix with 440.5 μL of KRAS nucleotide exchange buffer, incubate at room temperature for 1 hour, then mix with 8.4 μL of 17.9 μM Tb-SA, 1.8 μL of 5 mM Bodipy GDP, and 9539.8 μL of KRAS experimental buffer to prepare 1 L of solution, let it stand at room temperature for 6 hours after mixing, and store at -80 °C.

[0659] b. Experimental reagents:

[0660] 1) KRAS enzyme solution

[0661] Take 73.3 μL of KRAS / Bodipy GDP / Tb-SA mixture and 2126.7 μL of KRAS experimental buffer to prepare 2200 μL of solution.

[0662] 2) SOS / GTP mixture

[0663] c. Experimental procedure:

[0664] Take 1.59 μL of 166 μM SOS protein, 198 μL of 100 mM GTP, and 2000.41 μL of KRAS experimental buffer to prepare a 2200 μL solution. The stock concentration of the control compound is 1 mM, and the stock concentration of the test compound is 10 mM. Transfer 9 μL of the control compound and the test compound into a 384-LDV plate; use Bravo to perform a 10-point 3-fold dilution of the compounds on the LDV plate; use ECHO to transfer 9 nL of the compounds on the LDV plate to the experimental plate; use a Dragonfly automatic pipettor to add 3 μL of 3 nM Kras / 0.5 nM TB-SA / 30 nM 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 a Dragonfly automatic pipettor to add 3 μL of 120 nM SOS / 9 mM GTP mixture to each well of the experimental plate, centrifuge the experimental plate at 1000 rpm / min for 1 minute; incubate the experimental plate at room temperature for 1 hour; use Envision to read and record the data; use Excel and Xlfit for data analysis and calculate the IC of the test compound 50 where "++++" indicates IC 50 ≤ 5 nM; "+++" indicates 5 nM < IC 50 ≤ 50 nM; "++" indicates 50 nM < IC 50 ≤ 2000 nM; "+" indicates 2000 nM < IC 50 .

[0665] Table 1 IC50 values of the compounds for KRAS enzyme inhibition.

[0666]

[0667]

[0668] Experimental Example 2. Cell p-ERK Inhibition Test

[0669] Compounds that can effectively inhibit p-ERK of AsPC-1 (G12D), A549 (G12S), HCT116 (G13D), NCI-H358 (G12C), NCI-H460 (Q61H), NCI-H727 (G12V), MKN1 (WTdep), PSN-1 (G12R) cells were screened by the HTRF method.

[0670] Cells were seeded in a transparent 96-well cell culture plate, 80 μL of cell suspension per well, with 8000 cells per well. The cell plate was placed in a carbon dioxide incubator and incubated overnight at 37 °C. Take 2 μL of the compound and add it to 78 μL of cell culture medium. After mixing, take 20 μL of the compound solution and add it to the corresponding well of the cell plate. The cell plate was then returned to the carbon dioxide incubator and incubated for another 1 hour. After the incubation was completed, the cell supernatant was discarded and 50 μL of 1X cell lysis buffer was added to each well. Incubate with shaking at room temperature for 30 minutes. Dilute the Phospho-ERK1 / 2 Eu Cryptate antibody and Phospho-ERK1 / 2 d2 antibody 20-fold with detection buffer. Take 16 μL of the cell lysate supernatant from each well into a new 384-well white microplate, and then add 2 μL of the diluted Phospho-ERK1 / 2 Eu Cryptate antibody and 2 μL of the diluted Phospho-ERK1 / 2 d2 antibody. Incubate at room temperature for at least 4 hours. After the incubation was completed, use a multi-label analyzer to read HTRF excitation: 320 nm, emission: 615 nm, 665 nm;

[0671] Calculate the IC of the compound to be tested 50 。Among them, "++++" indicates IC 50 ≤10 nM; "+++" indicates 10 nM < IC 50 ≤100 nM; "++" indicates 100 nM < IC 50 ≤2000 nM; "+" indicates 2000 nM < IC 50 。

[0672] Table 2. IC of the compound's inhibition of p-ERK in tumor cells 50 (nM).

[0673]

[0674]

[0675] Although the present invention has been described in detail above, those skilled in the art understand 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 rights of the present invention is not limited to the detailed description above, but shall be attributed to the claims.

Claims

1. A compound, stereoisomer or pharmaceutically acceptable salt thereof having the formula (I): Each L1 is independently selected from a key, OC 0-6 alkyl, NHC 0-6 alkyl, C 1-6 alkyl, COC 0-6 alkyl or SC 0-6 alkyl; Each Ar is independently selected, at each occurrence, from 5- to 12-membered heteroaryl, the heteroaryl independently containing, at each occurrence, 1, 2, 3 or 4 heteroatoms selected from N, O, or S, wherein the heteroaryl is optionally substituted by one or more G 1 substituted; Each X1, independently upon each occurrence, is selected from N, CR4; Each of R2, R3, and R4 is independently selected from H, D, cyano, halogen, C 1-6 alkyl, CN; Each R1 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- to 10-membered heterocycle, 4- to 10-membered hetero-fused ring, 5- to 12-membered spiroheterocyclic group; the 3- to 10-membered heterocycle, 4- to 10-membered hetero-fused ring, and 5- to 12-membered spiroheterocyclic group each independently contain 1, 2, 3, or 4 heteroatoms selected from N, O, or S upon each occurrence; each R1 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 carbocyclic group, 3- to 10-membered heterocycle, either substituted or unsubstituted; U is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclic, 5-12 membered spirocyclic, 5-12 membered spiroheterocyclic, aryl or heteroaryl, each heterocycloalkyl, fused heterocyclic, spiroheterocyclic, heteroaryl independently contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S upon each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclic, fused ring, fused heterocyclic, spiroheterocyclic, aryl or heteroaryl is optionally substituted by one or more G 2 substituted; G 1 and G 2 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, -OR5, -OC(O)NR5R6, -C(O)OR5, -C(O)NR5R6, -C(O)R5, -NR5R6, -NR5C(O)R6, -NR5C(O)NR6R7, -S(O) i R5 or -NR5S(O) i R6, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl are optionally substituted by one or more deuterium, cyano, halogen, C 1-7 alkyl, C 2-7 alkenyl, C 2-7 alkynyl, C 3-9 cycloalkyl or 3-9 membered heterocyclic group, C 7-10 aryl, 6-10 membered heteroaryl, -OR8, -OC(O)NR8R9, -C(O)OR8, -C(O)NR8R9, -C(O)R8, -NR8R9, -NR8C(O)R9, -NR8C(O)NR9R 10 , -S(O) i R8 or -NR8S(O) i R9 substituents; R5, R6, R7, R8, R9 and R 10 are each independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, or a 3- to 8-membered monocyclic heterocyclic group, monocyclic heteroaryl or phenyl; m and i are 1 or 2.

2. The compound or its pharmaceutically acceptable salt according to claim 1, wherein the compound is represented by formula (II-A), (II-B), (II-C) or (II-D):

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, Ar is selected from:

4. The compound or its pharmaceutically acceptable salt according to any one of claims 1, selected from:

5. The compound or its pharmaceutically acceptable salt according to any one of claim 4, selected from:

6. The compound according to any one of claims 1-5, or its optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, solvate, characterized in that, The pharmaceutically acceptable salts are selected from the group consisting of: potassium salts, sodium salts, magnesium salts, calcium salts, sulfates, hydrochlorides, phosphates, sulfonates, or carbonates.

7. A pharmaceutical composition, characterized in that, Comprising the compound according to any one of claims 1-6, or its optical isomers, pharmaceutically acceptable salts, prodrugs, deuterated derivatives, hydrates, solvates, and a pharmaceutically acceptable carrier.

8. Use of a compound according to any one of claims 1-6, or an optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, solvate thereof, characterized in that, For the preparation of a pharmaceutical composition for the treatment of diseases, disorders or conditions related to the activity or expression level of KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H.

9. The use according to claim 8, wherein The diseases, disorders or conditions are 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, cutaneous granular cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, throat cancer, breast cancer, prostate cancer, glioblastoma, 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, splenomegalic polycythemia, hypereosinophilic syndrome, multiple myeloma and other various solid tumors and hematological tumors.

Images