Preparation and use of pyrimidothiopyranone kras mutant protein inhibitor

Abstract

The present invention relates to a KRASG12D inhibitor and the use thereof. Specifically, provided in the present invention is a compound as shown in formula (I), and the definition of each substituent in the formula is as described in the description. In addition, the present invention further relates to a composition containing the inhibitor and the use thereof. The compound of the present invention has good tumor growth inhibitory activity, and has good safety.

Description

Preparation and application of pyrimidothiopyranone KRAS mutant protein inhibitors

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the entirety of Chinese patent application No. 202410133666.8, filed January 30, 2024. This application incorporates the entirety of the aforementioned Chinese patent application. Technical Field

[0003] The present invention belongs to the field of drug synthesis, and specifically relates to a novel KRAS G12D Inhibitors and their preparation methods and uses. Background Art

[0004] The present invention generally relates to novel compounds and methods for their preparation and use as KRAS inhibitors, for example, for the treatment of cancer.

[0005] RAS represents a group of closely related, monomeric, globular proteins of 189 amino acids (molecular weight 21 kDa) that are associated with the plasma membrane and bind to either GDP or GTP. RAS acts as a molecular switch. When RAS contains bound GDP, it is in a quiescent, or closed, state and "inactive." In response to exposure of cells to certain growth-promoting stimuli, RAS is induced to convert its bound GDP into GTP. Once bound to GTP, RAS is "switched on" and able to interact with and activate other proteins (their "downstream targets"). RAS proteins themselves have a very low intrinsic capacity and are unable to hydrolyze GTP back to GDP, thus remaining in their closed state. Turning off RAS requires external 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 GAPs or convert GTP back to GDP results in prolonged activation of the protein, leading to prolonged cellular signaling that allows cells to continue growing and dividing. Because these signals drive cell growth and division, overactive RAS signaling may ultimately lead to cancer.

[0006] Structurally, the RAS protein contains a G domain, which is responsible for the enzymatic activity of RAS - guanine nucleotide binding and hydrolysis (GTPase reaction). It also contains a C-terminal extension called the CAAX box, which can be post-translationally modified and is responsible for targeting the protein to the membrane. The G domain is approximately 21-25 kDa in size and contains a phosphate-binding loop (P-loop). The P-loop is the pocket where nucleotides bind in the protein. This is a rigid part of the domain with conserved amino acid residues (glycine 12, threonine 26, and lysine 16) and is crucial for nucleotide binding and hydrolysis. The G domain also contains the so-called SwitchI (residues 30-40) and SwitchII (residues 60-76) regions, both of which are dynamic parts of the protein and are often referred to as "spring-loaded" mechanisms due to their ability to switch between resting and loaded states. The key interaction is a hydrogen bond formed by threonine 35 and glycine 60 with the γ-phosphate of GTP, which holds the Switch1 and Switch2 regions, respectively, in their active conformations. Upon GTP hydrolysis and release of the phosphates, the two relax to the inactive GDP conformation.

[0007] The best-known members of the RAS subfamily are HRAS, KRAS, and NRAS, primarily due to their association with various types of cancer. Mutations in any of the three major RAS isoforms (HRAS, NRAS, or KRAS) are the most common in human tumorigenesis. Approximately 30% of human tumors are found to harbor RAS gene mutations. Notably, KRAS mutations are detected in 25-30% of tumors. In contrast, the prevalence of oncogenic mutations in NRAS and HRAS family members is much lower (8% and 3%, respectively). The most common KRAS mutations are found at residues G12 and G13 of the P-loop and residue Q61. G12C is a frequent mutation in the KRAS gene (glycine 12 to cysteine). This mutation has been found in approximately 13% of cancers, approximately 43% of lung cancers, and approximately 100% of cases of MYH-associated polyposis (a familial colon cancer syndrome).

[0008] As a cutting-edge target, KRAS mutant protein has received widespread attention. Among them, AMG-510 developed by Amgen was approved for marketing by the FDA last year. In recent years, other companies have applied for a number of patents on KRAS inhibitors, such as WO2016164675, WO2016168540, WO2021141628, WO2022098625, WO2022087371, WO2020101736, WO2022109485, WO2022109487 and WO2020146613. Therefore, despite progress in this field, there is still a need in the art for improved compounds and methods for treating cancer, such as by inhibiting KRAS, HRAS or NRAS. The present invention meets this need and provides other related advantages.

[0009] In short, the present invention provides compounds capable of inhibiting KRAS mutations, including stereoisomers, pharmaceutically acceptable salts, tautomers and prodrugs thereof. Summary of the Invention

[0010] A compound represented by general formula (I), a stereoisomer, a pharmaceutically acceptable salt, or an isomer thereof, wherein the compound represented by general formula (I) has the following structure:

[0011] Each L1 is independently selected at each occurrence from a bond, an OC 0-6 Alkyl, NHC 0-6 Alkyl, C 1-6 Alkyl, COC 0-6 Alkyl or SC 0-6 alkyl;

[0012] Each Ar is independently selected at each occurrence from a 5-12 membered heteroaromatic group, wherein the heteroaromatic group independently at each occurrence contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S, wherein the heteroaromatic group is optionally substituted by one or more G 1 replaced by;

[0013] Each X1 is independently selected at each occurrence 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 3-6carbocyclyl, 3-10 membered heterocyclic ring, 4-10 membered heterocondensed ring, 5-12 membered spiro heterocyclic ring; 3-10 membered heterocyclic ring, 4-10 membered heterocondensed ring, 5-12 membered spiro heterocyclic ring, each occurrence of which independently contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S; each R1 is independently optionally replaced by 1, 2, 3, 4, 5 or 6 heteroatoms selected from deuterium, halogen, C 1-6 Alkyl, -C 1-6 Alkoxy, oxo, OC 1-6 Alkyl, C 3-6 The carbocyclic group and the 3-10 membered heterocyclic group may be substituted or unsubstituted;

[0016] U is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spirocyclyl, 5-12 membered spiroheterocyclyl, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclyl, spiroheterocyclyl, heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclyl, fused cyclyl, fused heterocyclyl, spiroheterocyclyl, aromatic or heteroaromatic is optionally substituted by one or more G 2 replaced by;

[0017] G 1 , G 2 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 heteroaromatic, -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 with one or more deuterium, cyano, halogen, C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkynyl, C 3-9 Cycloalkyl or 3-9 membered heterocyclic group, C 7-10 Aryl, 6-10 membered heteroaromatic, -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) iis substituted by a substituent of R9;

[0018] R5, R6, R7, R8, R9 and R 10 Each independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclic group, monocyclic heteroaromatic group or phenyl group;

[0019] m,i is 1 or 2.

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

[0021] In some embodiments, the compound of formula (I), its pharmaceutically acceptable salt or stereoisomer thereof, formula (I) is further represented by (II-A), (II-B), (II-C), (II-D), (II-E) or (II-F)

[0022] In some embodiments, the compound of formula (I) or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof are selected from the following compounds, their isomers, solvates or precursors, or pharmaceutically acceptable salts thereof:

[0023] In some embodiments, the compound of formula (I) or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof are selected from the following compounds, their isomers, solvates or precursors, or pharmaceutically acceptable salts thereof:

[0024] On the other hand, the present invention also provides a pharmaceutical composition comprising the compounds represented by formula (I) and formula (II) or pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients.

[0025] On the other hand, the present invention relates to a method for treating a disease associated with KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H in a mammal, comprising administering a therapeutically effective amount of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to a mammal, preferably a human, in need of such treatment.

[0026] On the other hand, the present invention relates to the use of compounds represented by formula (I) and formula (II) or pharmaceutically acceptable salts thereof in drugs 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.

[0027] On the other hand, the present invention relates to compounds represented by formula (I) and formula (II) or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof for preventing or treating diseases related to KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H.

[0028] Some chemical terms

[0029] Unless stated to the contrary, the following terms are used in the specification and claims.

[0030] The notation "C" used in this document has the following meanings: x-y " represents the range of carbon atoms, where x and y are both integers, for example, C 3-8 Cycloalkyl means a cycloalkyl group having 3 to 8 carbon atoms, i.e. a cycloalkyl group having 3, 4, 5, 6, 7 or 8 carbon atoms. It is also understood that “C 3-8 " also includes any sub-ranges therein, such as C 3-7 、C 3-6 、C 4-7 、C 4-6 、C 5-6 wait.

[0031] "Alkyl" refers to a straight or branched hydrocarbon group containing 1 to 20 carbon atoms, for example 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 group can be substituted or unsubstituted.

[0032] "Alkenyl" refers to a straight or branched hydrocarbon group containing at least one carbon-carbon double bond and typically 2 to 20 carbon atoms, such as 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkenyl include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 1,4-pentadienyl, and 1,4-butadienyl. The alkenyl group can be substituted or unsubstituted.

[0033] "Alkynyl" refers to a straight or branched hydrocarbon group containing at least one carbon-carbon triple bond and typically 2 to 20 carbon atoms, such as 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkynyl include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl. The alkynyl group can be substituted or unsubstituted.

[0034] "Cycloalkyl" refers to a saturated cyclic hydrocarbon substituent containing from 3 to 14 carbon ring atoms. A cycloalkyl group can be a single carbon ring, typically containing from 3 to 7 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. A cycloalkyl group can alternatively be a bicyclic or tricyclic ring fused together, such as decahydronaphthyl. The cycloalkyl group can be substituted or unsubstituted.

[0035] "Heterocyclyl," "heterocycloalkyl," and "heterocycle" refer to stable, 3- to 18-membered, monovalent, non-aromatic rings containing 2-12 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, heterocyclyl groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which may include fused, spirocyclic, or bridged ring systems. The nitrogen, carbon, or sulfur atoms in the heterocyclyl group may be optionally oxidized, the nitrogen atom may be optionally quaternized, and the heterocyclyl group may be partially or fully saturated. The heterocyclyl group may be attached to the rest of the molecule through a single bond via a carbon or heteroatom in the ring. Heterocyclyl groups containing fused rings may contain one or more aromatic or heteroaromatic rings, as long as the attachment to the rest of the molecule is to an atom in a non-aromatic ring. For the purposes of the present application, the heterocyclic group is preferably a stable 4-11 membered monovalent non-aromatic monocyclic or bicyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 4-8 membered monovalent non-aromatic monocyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heterocyclic groups include azepanyl, azetidinyl, decahydroisoquinolinyl, dihydrofuranyl, dihydroindolinyl, dioxolane, 1,1-dioxo-thiomorpholinyl, imidazolidinyl, imidazolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidonyl, pyranyl, pyrazolidinyl, pyrrolidinyl, quinolizinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydropyranyl, and the like.

[0036] "Spiro heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, wherein the single rings share an atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen or S (O) m (wherein m is an integer 0 to 2) heteroatoms, and all the other ring atoms are carbon. These can contain one or more double bonds, but no ring has a completely conjugated electronic system that is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of shared spiral atoms between ring and ring, spiroalkyl is divided into single spiral heterocyclic radical, double spiral heterocyclic radical or multi-spiro heterocyclic radical, preferably single spiral alkyl and double spiral alkyl. More preferably 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral alkyl. The non-limiting examples of spiro heterocyclic radical include:

[0037] "Fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, wherein each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m(wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic alkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

[0038] "Aryl" or "aromatic group" refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10 members, such as phenyl and naphthyl, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring.

[0039] "Heteroaryl" or "heteroaromatic" refers to a 5-16 membered ring system containing 1-15 carbon atoms, preferably 1-10 carbon atoms, 1-4 heteroatoms selected from nitrogen, oxygen, and sulfur, and at least one aromatic ring. Unless otherwise specified, a heteroaryl group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and may include fused or bridged ring systems, as long as the point of attachment to the rest of the molecule is an aromatic ring atom. Nitrogen atoms, carbon atoms, and sulfur atoms in the heteroaryl ring can be selectively oxidized, and nitrogen atoms can be selectively quaternized. For purposes of the present invention, a heteroaryl group is preferably a stable 4-11 membered monoaromatic ring containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, and more preferably a stable 5-8 membered monoaromatic ring containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heteroaryl groups include acridinyl, azepine, benzimidazolyl, benzindolyl, benzodioxinyl, benzodioxolyl, benzofuranonyl, benzofuranyl, benzonaphthofuranyl, benzopyrone, benzopyranyl, benzopyrazolyl, benzothiadiazolyl, benzothiazolyl, benzotriazolyl, furanyl, imidazolyl, indazolyl, indolyl, oxazolyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinuclyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazinyl, etc. In the present application, heteroaryl is preferably a 5-8 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably pyridinyl, pyrimidinyl, thiazolyl. The heteroaryl group may be substituted or unsubstituted.

[0040] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0041] "Hydroxy" refers to -OH, "amino" refers to -NH2, "amido" refers to -NHCO-, "cyano" refers to -CN, "nitro" refers to -CN, "isocyano" refers to -NC, and "trifluoromethyl" refers to -CF3.

[0042] The term "heteroatom" or "hetero" as used herein, alone or as part of another component, refers to atoms other than carbon and hydrogen, and 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.

[0043]

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

[0044]

[00146] The term "spiro" or "spirocycle," as used herein, alone or in combination, refers to a cyclic structure in which two or more rings share one or more atoms.

[0045] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and where it does not. For example, "a heterocycle group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocycle group is substituted with an alkyl group and instances where the heterocycle group is not substituted with an alkyl group.

[0046] "Substituted" means that one or more atoms, preferably 5, more preferably 1 to 3 atoms, in a group are independently substituted by a corresponding number of substituents. It goes without saying that the substituents are in their possible chemical positions and that those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, a free amino 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, etc.

[0047] A "pharmaceutical composition" refers to a composition containing one or more compounds described herein, or pharmaceutically acceptable salts or prodrugs thereof, and other components such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient, and thereby exert biological activity.

[0048] "Isomers" refer to compounds that have the same molecular formula but differ in the nature or order of their atomic bonding or the spatial arrangement of their atoms. Isomers whose atoms are arranged differently in space are called "stereoisomers." Stereoisomers include optical isomers, geometric isomers, and conformational isomers. The compounds of the present invention may exist as optical isomers. Depending on the configuration of substituents around the chiral carbon atom, these optical isomers are in the "R" or "S" configuration. Optical isomers include enantiomers and diastereomers. Methods for preparing and separating optical isomers are known in the art.

[0049] The compounds of the present invention may 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 rings. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as Z or E configurations, and substituents around cycloalkyl groups or heterocyclic rings are designated as cis or trans configurations.

[0050] The compounds of the present invention may also exhibit tautomerism, such as keto-enol tautomerism.

[0051] It is to be understood that the present invention encompasses any tautomeric or stereoisomeric forms and mixtures thereof and is not intended to be limited to any one tautomeric or stereoisomeric form used in the naming of the compounds or chemical formulas.

[0052] "Isotopes" are all isotopes of atoms that occur in the compounds of the present invention. Isotopes include those atoms having 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 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 accompanying examples using appropriate isotopically labeled reagents in place of non-isotopically labeled reagents. Such compounds have various potential uses, for example, as standards and reagents in assays for biological activity. In the case of stable isotopes, such compounds have the potential to advantageously alter biological, pharmacological, or pharmacokinetic properties.

[0053] "Prodrug" means that the compound of the present invention can be administered in the form of a prodrug. Prodrug refers to a derivative that is converted into a biologically active compound of the present invention under physiological conditions in vivo, for example, by oxidation, reduction, hydrolysis, etc. (each of which is carried out using an enzyme or without the participation of an enzyme). Examples of prodrugs are compounds wherein the amine group in the compound of the present invention is acylated, alkylated, or phosphorylated, such as eicosanoylamino, alanylamide, pivaloyloxymethylamino, or wherein the hydroxyl group is acylated, alkylated, phosphorylated, or converted into a borate, such as acetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaryloxy, alanyloxy, or wherein the carboxyl group is esterified or amidated, or wherein the sulfhydryl 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.

[0054] "Pharmaceutically acceptable salts" or "pharmaceutically acceptable" refers to salts prepared from pharmaceutically acceptable bases or acids, including inorganic bases or acids and organic bases or acids. 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, compounds of the present invention containing acidic groups may exist in salt form and 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, and the like, for example, ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purines, piperazine, piperidine, choline, and caffeine. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. The compound of the present invention that contains basic group can exist in salt form and can be used according to the present invention with the form of addition of them and inorganic or organic acid.The example of suitable acid comprises hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, phosphoric 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 acid well known to persons skilled in the art.If the compound of the present invention contains acidic and basic group simultaneously in molecule, the present invention also comprises inner salt or betaine except mentioned salt form.Each salt is obtained by conventional method well known to persons skilled in the art, for example, by making these and organic or inorganic acid or alkali contact or by with other salt anion exchange or cation exchange in solvent or dispersant.

[0055] Therefore, when referring to a "compound," "compound of the present invention," or "compound of the present invention" in this application, all forms of the compound, such as prodrugs, stable isotopic derivatives, pharmaceutically acceptable salts, isomers, meso- and racemates, enantiomers, diastereomers, and mixtures thereof, are included.

[0056] As used herein, the term "tumor" includes benign tumors and malignant tumors (eg, cancer).

[0057] As used herein, the term "cancer" includes various malignant tumors in which KRAS is involved, including but not limited to pancreatic cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, cutaneous granular cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, pharyngeal cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, skin cancer, lymphoma, gastric cancer, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, monocytic leukemia, splenomegaly, multiple eosinophilia syndrome, myeloma and other solid tumors and blood tumors.

[0058] As used herein, the terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" refer to an amount of at least one agent or compound sufficient to alleviate to some extent one or more symptoms of the disease or condition being treated. The result can be a reduction and / or alleviation of signs, symptoms, or causes of disease, or any other desired change in a biological system. For example, a therapeutically effective amount is the amount of a composition comprising a compound disclosed herein that provides a clinically significant alleviation of symptoms. Techniques such as dose escalation studies can be used to determine the effective amount appropriate for any individual case.

[0059] The term "polymorph" or "polymorphism" used in the present invention refers to the compounds of the present invention having multiple crystal lattice forms. Some compounds of the present invention may have more than one crystal form. The present invention covers all multiple forms or mixtures thereof.

[0060] Intermediate compounds of the compounds of the present invention and their polymorphs are also within the scope of the present invention.

[0061] Crystallization often produces solvates of the compounds of the present invention. The term "solvate" as used herein refers to an association of one or more molecules of the compound of the present invention with one or more solvent molecules.

[0062] The solvent may be water, in which case the solvate is a hydrate. Alternatively, it may be an organic solvent. Thus, the compounds of the present invention may exist as hydrates, including monohydrates, dihydrates, hemihydrates, trihydrates, tetrahydrates, and the like, as well as the corresponding solvated forms. While the compounds of the present invention may be true solvates, in other cases, the compounds of the present invention may simply retain water or a mixture of water and some other solvent. The compounds of the present invention may react in a solvent or precipitate or crystallize in a solvent. Solvates of the compounds of the present invention are also encompassed within the scope of the present invention.

[0063] The term "acceptable" with respect to a formulation, composition or ingredient, as used herein, means having no persistent detrimental effect on the general health of the subject being treated.

[0064] 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, that is, the substance can be administered to a subject without causing an adverse biological response or interacting in an adverse manner with any components contained in the composition.

[0065] "Pharmaceutically acceptable carrier" includes, but is not limited to, adjuvants, carriers, excipients, auxiliary agents, 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 relevant government administrative departments for use in humans and domesticated animals.

[0066] As used herein, the terms "subject," "patient," "subject," or "individual" refer to individuals suffering from a disease, disorder, or condition, including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, and pigs; domestic animals, such as rabbits, dogs, and cats; and laboratory animals, including rodents, such as rats, mice, and guinea pigs. Examples of non-human mammals include, but are not limited to, birds and fish. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0067] The term "treatment" as used herein refers to the treatment of a disease or condition in a mammal, especially a human, including

[0068] (i) preventing the development of a disease or condition in a mammal, particularly a mammal that has been previously exposed to the disease or condition but has not yet been diagnosed with the disease or condition;

[0069] (ii) inhibiting the disease or condition, i.e., controlling its development;

[0070] (iii) alleviate the disease or condition, i.e., cause the disease or condition to regress;

[0071] (iv) Alleviate symptoms caused by a disease or condition.

[0072] As used herein, the terms "disease" and "disorder" are used interchangeably or may have different meanings because certain diseases or disorders do not yet have a known causative agent (and therefore the cause of the disease is unknown) and therefore cannot be considered diseases but rather are considered to be undesirable conditions or syndromes with more or less specific symptoms that have been confirmed by clinical researchers.

[0073] As used herein, the terms "administer," "administer," "administer," and the like refer to methods that enable a compound or composition to be delivered to the desired site of biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

[0074] Specific implementation methods

[0075] The present invention also provides a method for preparing the compound. The preparation of the compound described in the general formula (I) of the present invention can be completed by the following exemplary methods and examples, but these methods and examples should not be considered to limit the scope of the present invention in any way. The compound described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of methods known in the art and the method described in the present invention can be used. The product obtained in each step is obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized or purchased according to the literature.

[0076] Unless otherwise stated, temperatures are in degrees Celsius. Reagents were purchased from commercial suppliers such as Chemblocks Inc and 3wpharm and were used without further purification unless otherwise stated.

[0077] Unless otherwise stated, the following reactions were performed at room temperature in anhydrous solvents under a positive pressure of nitrogen or CO or using a drying tube; glassware was oven-dried and / or heat-dried.

[0078] Unless otherwise specified, column chromatography purification used 200-300 mesh silica gel from Qingdao Ocean Chemical Plant; preparative thin-layer chromatography silica gel precast plates (HSGF254) produced by Yantai Chemical Industry Research Institute were used; and MS was determined using a Therno LCD Fleet (ESI) liquid chromatography-mass spectrometer.

[0079] Nuclear magnetic resonance data (1H NMR) were obtained using a Bruker Avance-400 MHz or Varian Oxford-400 Hz NMR spectrometer. The solvents used for the NMR data included CDCl3, CD3OD, D2O, DMS-d6, etc., and the data were based on tetramethylsilane (0.000 ppm) or residual solvent (CDCl 3: When peak diversity is indicated, the following abbreviations are used to denote the different peak shapes: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad), dd (double of doublets), dt (double of triplets). Coupling constants, if given, are in Hertz (Hz).

[0080] Preparation of intermediates

[0081] Preparation of A1

[0082] Step A

[0083] In a dry 1L 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. The temperature was cooled to 0°C. Under nitrogen, a solution of compound A1-1 (10 g, 80 mmol) in N,N-dimethylformamide (200 mL) was added dropwise. The reaction was continued at 0°C for 0.5 hours. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20°C, and stirring was continued under nitrogen for 7.5 hours. The reaction solution was slowly added to 100 mL of saturated ammonium chloride and extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to yield compound A1-2. LC-MS (ESI): m / z = 365.45 [M+H] + .

[0084] Step B

[0085] 2,2.6,6-Tetramethylpiperidine (11.4 g, 82.1 mmol) was added to anhydrous tetrahydrofuran (500 mL), cooled to -5°C, n-butyllithium (2.5 M, 40 mL) was added dropwise, and the mixture was reacted at -5 to 0°C for 15 minutes. The mixture was cooled to -60°C, and a solution of compound A1-2 (10 g. 27 mmol) in tetrahydrofuran (30 mL) was added. The mixture was reacted at -60°C for 0.5 hours. N,N-dimethylformamide (40 g, 0.5 mol) was quickly added, and the reaction solution was stirred at -60°C for 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was slurried with the solvent for 0.5 hours, filtered, and the filter cake dried. The filtrate was concentrated and purified by silica gel column chromatography. The filter cake and column chromatography were combined to obtain compound A1-3. LC-MS (ESI): m / z = 393.5 [M+H] + .

[0086] Step C

[0087] Compound A1-3 (10 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and bromosuccinimide (4.5 g, 25 mmol) was added. The reaction solution was stirred at 20°C for 20 minutes. The reaction solution 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 A1-4. LC-MS (ESI): m / z = 472.4 [M+H] + .

[0088] Step D

[0089] Compound A1-4 (9.7 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Cuprous iodide (7.8 g, 41 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 celite. 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 A1-5. LC-MS (ESI): m / z = 461.5 [M+H] + .

[0090] Step E

[0091] Under nitrogen, 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. Dimethyl ethylphosphoryl acetate (3.55 g, 19.5 mmol) was then added. The reaction solution was stirred at room temperature for 24 hours. The THF was recovered and added to 150 mL of water. The mixture was extracted with methyl tert-butyl ether (75 mL x 2). The combined organic phases were washed with saturated brine (100 mL x 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 A1-6. LC-MS (ESI): m / z = 531.6 [M+H] + .

[0092] Step F

[0093] 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 the mixture was extracted with dichloromethane (75 mL x 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0094] Step G

[0095] Compound A1-7 (7.98 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. The mixture was poured into 100 mL of water and extracted with dichloromethane (100 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0096] Step H

[0097] To a suspension of A1-8 (3.3 g, 5.3 mmol) in water (10 mL) were 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 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. 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] + .

[0098] Step I

[0099] Compound A1-9 (1 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and 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 mixture. The mixture was allowed to react at this temperature for 15 minutes. The reaction mixture was poured into saturated aqueous ammonium chloride (8 mL), and the layers were separated. The aqueous phase was extracted with dichloromethane (5 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain compound A1. LC-MS (ESI): m / z = 777.8 [M+H] + .

[0100] Preparation of B1

[0101] Step A

[0102] To a solution of B1-1 (25.3 g, 201 mmol) in N,N-dimethylformamide (500 mL) was added bromomethylbenzene (41.2 g, 241 mmol) and potassium carbonate (55.5 g, 402 mmol). After stirring at 25 ° C for 15 hours, 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. Purification by silica gel column chromatography gave B1-2. LC-MS (ESI): m / z = 216.3 [M + H] + .

[0103] Step B

[0104] 2,2.6,6-Tetramethylpiperidine (11.4 g, 82.1 mmol) was added to anhydrous tetrahydrofuran (500 mL), cooled to -5°C, n-butyllithium (2.5 M, 40 mL) was added dropwise, and the mixture was reacted at -5 to 0°C for 15 minutes. The mixture was cooled to -60°C, and a solution of compound B1-2 (5.8 g, 27 mmol) in tetrahydrofuran (30 mL) was added. The mixture was reacted at -60°C for 0.5 hours. N,N-dimethylformamide (40 g, 0.5 mol) was quickly added, and the reaction solution was stirred at -60°C for 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was slurried with the solvent for 0.5 hours, filtered, and the filter cake dried. The filtrate was concentrated and purified by silica gel column chromatography. The filter cake and column chromatography were combined to obtain compound B1-3. LC-MS (ESI): m / z = 245.1 [M+H] + .

[0105] Step C

[0106] Compound B1-3 (6.1 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and bromosuccinimide (4.5 g, 25 mmol) was added. The reaction solution was stirred at 20°C for 20 minutes. The reaction solution 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 hours, filtered, and the filter cake was dried to obtain compound B1-4. LC-MS (ESI): m / z = 323.2 [M+H] + .

[0107] Step D

[0108] Compound B1-4 (6.57 g, 20.4 mmol) was added to N,N-dimethylformamide (95 mL). Cuprous iodide (7.8 g, 41 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 celite. 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 B1-5. LC-MS (ESI): m / z = 313.3 [M+H] + .

[0109] Step E

[0110] Under nitrogen, 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, followed by the addition of dimethyl ethylphosphoryl acetate (3.55 g, 19.5 mmol). The reaction solution was stirred at room temperature for 24 hours. The THF was recovered and added to 150 mL of water. 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 solvent was removed under reduced pressure. The resulting 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] + .

[0111] Step F

[0112] 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 mixture was stirred at 60°C for 48 hours. Dilute hydrochloric acid was added, and the mixture was extracted with dichloromethane (75 mL x 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0113] Step G

[0114] Compound B1-7 (6.2 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. The mixture 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 resulting 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] + .

[0115] Step H

[0116] To a suspension of B1-8 (2.49 g, 5.3 mmol) in water (10 mL) were 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 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. The resulting crude product was separated and purified by silica gel column chromatography to obtain compound B1-9. LC-MS (ESI): m / z = 497.1 [M+H] + .

[0117] Step I

[0118] Compound B1-9 (0.79 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and 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 allowed to react at this temperature for 15 minutes. The reaction solution was poured into saturated aqueous ammonium chloride (8 mL), and the layers were separated. The aqueous phase was extracted with dichloromethane (5 mL x 2). The combined organic phases were 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 B1. LC / MS (ESI): m / z = 629.0 [M+H] + .

[0119] Preparation of C1

[0120] Compound C1 was obtained using a similar preparation method to Intermediate A1 (the starting material was replaced with 6-bromo-4-methylpyridin-2-amine). LC / MS (ESI): m / z = 760.8 [M+H] + .

[0121] Preparation of D1

[0122] Compound D1 was obtained using a similar preparation method to intermediate B1 (the starting material was replaced with 6-bromo-4-methylpyridin-2-ol). LC / MS (ESI): m / z = 611.6 [M+H] + .

[0123] Preparation of E1

[0124] Step A

[0125] In a dry 1L 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. The reaction was continued at 0°C for 0.5 hours. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20°C, and stirring was continued under nitrogen for 7.5 hours. The reaction solution was slowly added to 100 mL of saturated ammonium chloride and extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to yield compound E1-2. LC-MS (ESI): m / z = 430.1 [M+H] + .

[0126] Step B

[0127] Compound E1-2 (34.6 g, 80.7 mmol) was added 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). The mixture was reacted at 120°C under nitrogen for 2 hours. 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] + .

[0128] Step C

[0129] 2,2.6,6-Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL), cooled to -5°C, n-butyllithium (2.5 M, 50 mL) was added dropwise, and the mixture was reacted at -5 to 0°C for 15 minutes. The mixture was cooled to -60°C, and a solution of compound E1-3 (24.27 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The mixture was reacted at -60°C for 0.5 hours. N,N-dimethylformamide (22.8 g, 0.312 mol) was quickly added, and the reaction solution was stirred at -60°C for 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was slurried with the solvent for 0.5 hours, filtered, and the filter cake dried. The filtrate was concentrated and purified by silica gel column chromatography. The filter cake and column chromatography were combined to obtain compound E1-4. LC-MS (ESI): m / z = 418.1 [M+H] + .

[0130] Step D

[0131] Compound E1-4 (10.43 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and bromosuccinimide (4.5 g, 25 mmol) was added. The reaction solution was stirred at 20°C for 20 minutes. The reaction solution 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] + .

[0132] Step E

[0133] 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 celite. 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] + .

[0134] Step E

[0135] Under nitrogen, 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, followed by the addition of dimethyl ethylphosphoryl acetate (3.55 g, 19.5 mmol). The reaction solution was stirred at room temperature for 24 hours. The THF was recovered and added to 150 mL of water. 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 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] + .

[0136] Step F

[0137] 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 x 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography to yield compound E1-7. LC-MS (ESI): m / z = 690.3 [M+H] + .

[0138] Step G

[0139] Compound E1-8 (7.97 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. The mixture was poured into 100 mL of water and extracted with dichloromethane (100 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0140] Step H

[0141] To a suspension of E1-9 (3.28 g, 5.3 mmol) in water (10 mL) were 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 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. The resulting 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] + .

[0142] Step I

[0143] Compound E1-10 (1.03 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and 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 mixture. The mixture was allowed to react at this temperature for 15 minutes. The reaction mixture was poured into saturated aqueous ammonium chloride (8 mL), and the layers were separated. The aqueous phase was extracted with dichloromethane (5 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain compound E1. LC-MS (ESI): m / z = 802.1 [M+H] + .

[0144] Preparation of F1

[0145] Step A

[0146] In a dry 1L 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 (23.76 g, 80 mmol) in N,N-dimethylformamide (500 mL) was added dropwise under nitrogen. The reaction was continued at 0°C for 0.5 hours. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20°C, and stirring was continued under nitrogen for 7.5 hours. The reaction solution was slowly added to 100 mL of saturated ammonium chloride and extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to yield compound F1-2. LC-MS (ESI): m / z = 567.9 [M+H] + .

[0147] Step B

[0148] Compound F1-2 (45.67 g, 80.7 mmol) was added 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). The mixture was reacted at 120°C under nitrogen for 2 hours. The reaction solution was concentrated under reduced pressure and separated by column chromatography to obtain compound F1-3. LC-MS (ESI): m / z = 451 [M+H] + .

[0149] Step C

[0150] 2,2.6,6-Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL), cooled to -5°C, n-butyllithium (2.5 M, 50 mL) was added dropwise, and the mixture was reacted at -5 to 0°C for 15 minutes. The mixture was cooled to -60°C, and a solution of compound E1-3 (28.08 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The mixture was reacted at -60°C for 0.5 hours. N,N-dimethylformamide (22.8 g, 0.312 mol) was quickly added, and the reaction solution was stirred at -60°C for 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was slurried with the solvent for 0.5 hours, filtered, and the filter cake dried. The filtrate was concentrated and purified by silica gel column chromatography. The filter cake and column chromatography were combined to obtain compound F1-4. LC-MS (ESI): m / z = 400.2 [M+H] + .

[0151] Step D

[0152] Compound F1-4 (10.43 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and bromosuccinimide (4.5 g, 25 mmol) was added. The reaction solution was stirred at 20°C for 20 minutes. The reaction solution was added to 150 mL of water and extracted with methyl tert-butyl ether (80 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 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 F1-5. LC-MS (ESI): m / z = 478.0 [M+H] + .

[0153] Step E

[0154] Compound F1-5 (9.77 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 celite. The filtrate was added to 150 mL of water and extracted with methyl tert-butyl ether (75 mL x 2). The combined organic phases were washed with saturated brine (100 mL x 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 F1-6. LC-MS (ESI): m / z = 468.1 [M+H] + .

[0155] Step E

[0156] Under nitrogen, 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, followed by the addition of dimethyl ethylphosphoryl acetate (3.55 g, 19.5 mmol). The reaction solution was stirred at room temperature for 24 hours. The THF was recovered and added to 150 mL of water. 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 solvent was removed under reduced pressure. The resulting 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] + .

[0157] Step F

[0158] 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% 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 x 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography to yield compound F1-8. LC-MS (ESI): m / z = 670.1 [M+H] + .

[0159] Step G

[0160] Compound F1-8 (8.39 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. The mixture was poured into 100 mL of water and extracted with dichloromethane (100 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0161] Step H

[0162] To a suspension of E1-9 (3.45 g, 5.3 mmol) in water (10 mL) were 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 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. 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] + .

[0163] Step I

[0164] Compound F1-10 (1.04 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and 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 mixture. The mixture was allowed to react at this temperature for 15 minutes. The reaction mixture was poured into saturated aqueous ammonium chloride (8 mL), and the layers were separated. The aqueous phase was extracted with dichloromethane (5 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain compound F1. LC-MS (ESI): m / z = 784.1 [M+H] + .

[0165] Preparation of G1

[0166] Step A

[0167] In a dry 1L 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. The reaction was continued at 0°C for 0.5 hours. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20°C, and stirring was continued under nitrogen for 7.5 hours. The reaction solution was slowly added to 100 mL of saturated ammonium chloride and extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to yield compound G1-2. LC-MS (ESI): m / z = 467.0 [M+H] + .

[0168] Step B

[0169] 2,2.6,6-Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL), cooled to -5°C, n-butyllithium (2.5 M, 50 mL) was added dropwise, and the mixture was reacted at -5 to 0°C for 15 minutes. The mixture was cooled to -60°C, and a solution of compound G1-2 (29.08 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The mixture was reacted at -60°C for 0.5 hours. N,N-dimethylformamide (22.8 g, 0.312 mol) was quickly added, and the reaction solution was stirred at -60°C for 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was slurried with the solvent for 0.5 hours, filtered, and the filter cake dried. The filtrate was concentrated and purified by silica gel column chromatography. The filter cake and column chromatography were combined to obtain compound G1-3. LC-MS (ESI): m / z = 396.1 [M+H] + .

[0170] Step D

[0171] Compound G1-3 (9.88 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and bromosuccinimide (4.5 g, 25 mmol) was added. The reaction solution was stirred at 20°C for 20 minutes. The reaction solution 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 G1-4. LC-MS (ESI): m / z = 474.0 [M+H] + .

[0172] Step E

[0173] Compound G1-4 (9.65 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 celite. 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 G1-5. LC-MS (ESI): m / z = 464.1 [M+H] + .

[0174] Step E

[0175] Under nitrogen, 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, followed by the addition of dimethyl ethylphosphoryl acetate (3.55 g, 19.5 mmol). The reaction solution was stirred at room temperature for 24 hours. The THF was recovered and added to 150 mL of water. 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 solvent removed under reduced pressure. The resulting 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] + .

[0176] Step F

[0177] 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 mixture was stirred at 60°C for 48 hours. Dilute hydrochloric acid was added, and the mixture was extracted from dichloromethane (75 mL x 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0178] Step G

[0179] Compound G1-7 (8.02 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. The mixture was poured into 100 mL of water and extracted with dichloromethane (100 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0180] Step H

[0181] To a suspension of G1-8 (3.29 g, 5.3 mmol) in water (10 mL) were 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 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. 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] + .

[0182] Step I

[0183] Compound G1-9 (1.04 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and 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 mixture. The mixture was allowed to react at this temperature for 15 minutes. The reaction mixture was poured into saturated aqueous ammonium chloride (8 mL), and the layers were separated. The aqueous phase was extracted with dichloromethane (5 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain compound G1. LC-MS (ESI): m / z = 780.1 [M+H] + .

[0184] Preparation of H1

[0185] Step A

[0186] In a dry 1L 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 H1-1 (14.88 g, 80 mmol) in N,N-dimethylformamide (500 mL) was added dropwise under nitrogen. The reaction was continued at 0°C for 0.5 hours. p-Methoxybenzyl chloride (27.5 g, 170 mmol) was added, the temperature was slowly raised to 20°C, and stirring was continued under nitrogen for 7.5 hours. The reaction solution was slowly added to 100 mL of saturated ammonium chloride and extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was purified by silica gel column chromatography to yield compound H1-2. LC-MS (ESI): m / z = 426.1 [M+H] + .

[0187] Step B

[0188] 2,2.6,6-Tetramethylpiperidine (22.3 g, 156 mmol) was added to anhydrous tetrahydrofuran (300 mL), cooled to -5°C, n-butyllithium (2.5 M, 50 mL) was added dropwise, and the mixture was reacted at -5 to 0°C for 15 minutes. The mixture was cooled to -60°C, and a solution of compound H1-2 (26.52 g, 62.4 mmol) in tetrahydrofuran (100 mL) was added. The mixture was reacted at -60°C for 0.5 hours. N,N-dimethylformamide (22.8 g, 0.312 mol) was quickly added, and the reaction solution was stirred at -60°C for 10 minutes. 200 mL of saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with methyl tert-butyl ether (100 mL x 2). The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was slurried with the solvent for 0.5 hours, filtered, and the filter cake dried. The filtrate was concentrated and purified by silica gel column chromatography. The filter cake and column chromatography were combined to obtain compound H1-3. LC-MS (ESI): m / z = 376.2 [M+H] + .

[0189] Step D

[0190] Compound H1-3 (9.34 g, 25 mmol) was added to 100 mL of N,N-dimethylformamide, and bromosuccinimide (4.5 g, 25 mmol) was added. The reaction solution was stirred at 20°C for 20 minutes. The reaction solution 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 H1-4. LC-MS (ESI): m / z = 454.0 [M+H] + .

[0191] Step E

[0192] Compound H1-4 (9.24 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 celite. 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 H1-5. LC-MS (ESI): m / z = 444.1 [M+H] + .

[0193] Step E

[0194] Under nitrogen, 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. Dimethyl ethylphosphoryl acetate (3.55 g, 19.5 mmol) was then added. The reaction solution was stirred at room temperature for 24 hours. The THF was recovered and added to 150 mL of water. The mixture was extracted with methyl tert-butyl ether (75 mL x 2). The combined organic phases were washed with saturated brine (100 mL x 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 H1-6. LC-MS (ESI): m / z = 514.1 [M+H] + .

[0195] Step F

[0196] 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 the mixture was extracted with dichloromethane (75 mL x 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography to yield compound H1-7. LC-MS (ESI): m / z = 648.3 [M+H] + .

[0197] Step G

[0198] 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. The mixture was poured into 100 mL of water and extracted with dichloromethane (100 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the organic solvent removed under reduced pressure. The resulting 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] + .

[0199] Step H

[0200] To a suspension of H1-8 (3.19 g, 5.3 mmol) in water (10 mL) were 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 hours. The precipitate was collected, washed with water (twice) and isopropyl ether, and dried. The resulting crude product was separated and purified by silica gel column chromatography to obtain compound H1-9. LC-MS (ESI): m / z = 628.2 [M+H] + .

[0201] Step I

[0202] Compound H1-9 (1.00 g, 1.6 mmol) was dissolved in dichloromethane (10 mL), and 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 mixture. The mixture was allowed to react at this temperature for 15 minutes. The reaction mixture was poured into saturated aqueous ammonium chloride (8 mL), and the layers were separated. The aqueous phase was extracted with dichloromethane (5 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain compound H1. LC-MS (ESI): m / z = 760.1 [M+H] + .

[0203] Example 1

[0204] 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)

[0205] Step A

[0206] Compound A1 (82 mg, 105.02 μmol) and 8-tert-butyloxycarbonyl-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. The reaction mixture was heated to 100°C and stirred for 1 hour. After cooling, the organic solvent was removed under reduced pressure. The crude product was purified by preparative thin-layer chromatography to obtain compound 1-1. LC / MS (ESI): m / z = 840 [M+H]. + .

[0207] Step B

[0208] Compound 1-1 (71 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperbenzoic 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 purified by preparative thin-layer chromatography to obtain compound 1-2. LC / MS (ESI): m / z = 904 [M+H] + .

[0209] Step C

[0210] In an ice-water bath, compound ((2R,7AS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (12.09 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 1-3. LC / MS (ESI): m / z = 983.1 [M+H] + .

[0211] Step D

[0212] 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 = 642.7 [M+H] + . 1 H-NMR(DMSO-d6)δ1.78-2.39(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,12H),5.96(s,2H),6.84(d,1H).

[0213] Example 2

[0214] (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)

[0215] (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)

[0216] Compound 1 was separated and purified by supercritical fluid 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] + ).

[0217] Example 3

[0218] 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)

[0219] Compound 2-4 was obtained using a similar preparation method to compound 1 in Example 1 (the starting material was replaced with B1). LC / MS (ESI): m / z = 733.8 [M+H] + .

[0220] Compound 2-4 (0.1 g, 1.4 mmol) was added to a 1:1 IPA:THF solution (25 mL) and stirred. 10% Pd / C (0.1 g) was then added to the stirred solution. The reaction mixture was stirred at room temperature under hydrogen for 16 h, 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] + .

[0221] Example 4

[0222] (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)

[0223] (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)

[0224] Compounds 2A and 2B were obtained using a similar preparation method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 2). LC / MS (ESI): m / z = 643.7 [M+H] + .

[0225] Example 5

[0226] 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)

[0227] Compound 3 was obtained by a similar preparation method to that of compound 1 in Example 1 (the starting material was replaced by C1). LC / MS (ESI): m / z = 625.7 [M+H] + . 1H-NMR(CDCl3)δ1.78-2.20(m,7H),2.01-2.20(m,6H),2.25-2.60(m,2H),3.00-3.20(m,3H) ,3.30-3.50(m,1H),3.60-3.80(m,2H),3.80-4.15(m,4H),4.30-4.98(m,7H),7.30(s,1H).

[0228] Example 6

[0229] (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)

[0230] (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)

[0231] Compounds 3A and 3B were obtained using a similar preparation method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 3). LC / MS (ESI): m / z = 625.7 [M+H] + .

[0232] Example 7

[0233] 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)

[0234] Compound 4 was obtained by a similar preparation method to that of compound 2 in Example 3 (the starting material was replaced with D1). LC / MS (ESI): m / z = 626.7 [M+H] + .

[0235] Example 8

[0236] (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)

[0237] (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)

[0238] Compounds 4A and 4B were obtained using a similar preparation method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 4). LC / MS (ESI): m / z = 626.7 [M+H] + .

[0239] Example 9

[0240] 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)

[0241] Compound 5 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 645.7 [M+H] + .

[0242] Example 10

[0243] (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)

[0244] (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)

[0245] Compounds 5A and 5B were prepared using a similar method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 5). LC / MS (ESI): m / z = 645.7 [M+H] + .

[0246] Example 11

[0247] 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)

[0248] Compound 6 was obtained by a similar preparation method to that of compound 2 in Example 3 (the starting material was replaced with B1). LC / MS (ESI): m / z = 646.7 [M+H] + .

[0249] Example 12

[0250] (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)

[0251] (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)

[0252] Compounds 6A and 6B were obtained using a similar preparation method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 6). LC / MS (ESI): m / z = 646.7 [M+H] + .

[0253] Example 13

[0254] 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)

[0255] Compound 7 was obtained by a similar preparation method to that of compound 1 in Example 1 (the starting material was replaced by C1). LC / MS (ESI): m / z = 628.7 [M+H] + .

[0256] Example 14

[0257] (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)

[0258] (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)

[0259] Compounds 7A and 7B were prepared using a similar method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 7). LC / MS (ESI): m / z = 628.7 [M+H] + .

[0260] Example 15

[0261] 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)

[0262] Compound 8 was obtained by a similar preparation method to that of compound 2 in Example 3 (the starting material was replaced with D1). LC / MS (ESI): m / z = 629.7 [M+H] + .

[0263] Example 16

[0264] (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)

[0265] (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)

[0266] Compounds 8A and 8B were obtained using a similar preparation method to that of compounds 1A and 1B in Example 2 (the starting material was replaced with compound 8). LC / MS (ESI): m / z = 629.7 [M+H] + .

[0267] Example 17

[0268] 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)

[0269] Step A

[0270] In an ice-water bath, ((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (12.09 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 1-3. LC / MS (ESI): m / z = 983.1 [M+H] + .

[0271] Step B

[0272] Compound 1-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 HPLC to obtain compound 17. LC / MS (ESI): m / z = 642.7 [M+H] + . 1 H-NMR(DMSO-d6)δ1.99(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),4.09-4.76(m,13H),5.96(s,2H)6.84(d,1H).

[0273] Example 18

[0274] (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)

[0275] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 9B)

[0276] Compound 9 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0277] Example 19

[0278] 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)

[0279] Step A

[0280] In an ice-water bath, ((S)-2-(Fluoromethylene)tetrahydro-1H-pyrrolizine-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, 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 10-1. LC / MS (ESI): m / z = 995.4 [M+H] + .

[0281] Step B

[0282] Compound 10-1 (43 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 10. LC / MS (ESI): m / z = 655.2 [M+H] + . 1H-NMR(DMSO-d6)δ1.68-2.31(m,12H),2.75(d,1H),2.94(d,1H),3.17-3.61(m,5H),4.05-4.78(m,12H),5.96(s,2H),6.84-7.05(m,2H).

[0283] Example 20

[0284] (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)

[0285] (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)

[0286] Compound 10 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0287] Example 21

[0288] 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)

[0289] Step A

[0290] In an ice-water bath, ((S)-2-(methylene)tetrahydro-1H-pyrrolizine-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, 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 11-1. LC / MS (ESI): m / z = 977.1 [M+H] + .

[0291] Step B

[0292] Compound 11-1 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 11. LC / MS (ESI): m / z = 637.3 [M+H] + . 1 H-NMR(DMSO-d6)δ1.79–2.31(m,12H),2.55-275(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).

[0293] Example 22

[0294] (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)

[0295] (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)

[0296] Compound 11 was separated and purified by supercritical fluid chromatography (SFC) to give 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 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 642.7 [M+H] + ).

[0297] Example 23

[0298] 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)

[0299] Compound 12 was obtained using a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 640.3 [M+H] + .

[0300] Example 24

[0301] (7S)-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 12A)

[0302] (7R)-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 12B)

[0303] Compounds 12A and 12B were prepared using a similar method to that used for compounds 2A and 2B in Example 2 (the starting material was replaced with compound 12). LC / MS (ESI): m / z == 640.3 [M+H] + .

[0304] Example 25

[0305] 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)

[0306] Compound 13 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 658.2 [M+H] + .

[0307] Example 26

[0308] (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)

[0309] (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)

[0310] Compounds 13A and 13B were prepared using a similar method to that used for compounds 2A and 2B in Example 2 (the starting material was replaced with compound 13). LC / MS (ESI): m / z = 658.2 [M+H] + .

[0311] Example 27

[0312] 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)

[0313] Compound 14 was obtained using a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 676.2 [M+H] + .

[0314] Example 28

[0315] (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)

[0316] (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)

[0317] Compounds 14A and 14B were prepared using a similar method to that used for compounds 2A and 2B in Example 2 (substituting compound 14 for the starting material). LC / MS (ESI): m / z = 676.2 [M+H] + .

[0318] Example 29

[0319] 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)

[0320] Compound 15 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 659.2 [M+H] + .

[0321] Example 30

[0322] (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)

[0323] (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)

[0324] Compounds 15A and 15B were prepared using a similar method to that of compounds 2A and 2B in Example 2 (the starting material was replaced with compound 15). LC / MS (ESI): m / z 659.2 [M+H] + .

[0325] Example 31

[0326] 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)

[0327] Compound 16 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 641.2 [M+H] + .

[0328] Example 32

[0329] (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)

[0330] (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)

[0331] Compounds 16A and 16B were prepared using a similar method to that used for compounds 2A and 2B in Example 2 (substituting compound 16 for the starting material). LC / MS (ESI): m / z = 641.2 [M+H] + .

[0332] Example 33

[0333] 7-(6-amino-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 17)

[0334] Compound 17 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 622.3 [M+H] + .

[0335] Example 34

[0336] (7S)-7-(6-amino-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 17A)

[0337] (7R)-7-(6-amino-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 17B)

[0338] Compounds 17A and 17B were prepared using a similar method to that used for compounds 2A and 2B in Example 2 (substituting compound 17 for the starting material). LC / MS (ESI): m / z = 622.3 [M+H] + .

[0339] Example 35

[0340] 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 18)

[0341] Step A

[0342] In an ice-water bath, ((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (14.28 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 18-1. LC / MS (ESI): m / z = 996.1 [M+H] + .

[0343] Step B

[0344] Compound 18-1 (43 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 18. LC / MS (ESI): m / z = 656.2 [M+H] + . 1 H-NMR(CDCl3)δ1.79–2.31(m,12H),2.75(d,1H),2.94(d,1H),3.17-3.61(m,5H),4.05-4.68(m,11H),7.30(s,1H).

[0345] Example 36

[0346] (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)

[0347] (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)

[0348] Compound 18 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0349] Example 37

[0350] 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)

[0351] Step A

[0352] In an ice-water bath, ((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizine-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, 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 1-3. LC / MS (ESI): m / z = 983.1 [M+H] + .

[0353] Step B

[0354] Compound 1-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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] + .

[0355] Example 38

[0356] (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)

[0357] (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)

[0358] Compound 19 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0359] Example 39

[0360] 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)

[0361] Step A

[0362] In an ice-water bath, ((S)-2-(methylene)tetrahydro-1H-pyrrolizine-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 20-1. LC / MS (ESI): m / z = 983.1 [M+H] + .

[0363] Step B

[0364] Compound 20-1 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 = 620.3 [M+H] + . 1 H-NMR(CDCl3)δ1.78–2.31(m,12H),2.55-75(m,1H),2.94(d,1H),3.17-3.61(m,5H),4.05-4.62(m,11H),5.20(d,2H),7.30(s,1H).

[0365] Example 40

[0366] (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)

[0367] (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)

[0368] Compound 20 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0369] Example 41

[0370] 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)

[0371] Step A

[0372] Compound E1 (84 mg, 105.02 μmol) and 8-tert-butyloxycarbonyl-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. The reaction mixture was heated to 100°C and stirred for 1 hour. After cooling, the organic solvent was removed under reduced pressure. The crude product was separated and purified by preparative thin-layer chromatography to obtain compound 21-1. LC / MS (ESI): m / z = 864 [M+H]. + .

[0373] Step B

[0374] Compound 21-1 (73 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperbenzoic 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 21-2. LC / MS (ESI): m / z = 928 [M+H] + .

[0375] Step C

[0376] In an ice-water bath, compound ((2R,7AS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (12.09 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 21-2 (60 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 21-3. LC / MS (ESI): m / z = 1007.4 [M+H] + .

[0377] Step D

[0378] Compound 21-3 (43 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 21. LC / MS (ESI): m / z = 667.2 [M+H] + . 1H-NMR(CDCl3)δ1.78-2.20(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),4.30-4.98(m,7H),6.82(d,1H).

[0379] Example 42

[0380] (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-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 21A)

[0381] (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)

[0382] 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] + ).

[0383] Example 43

[0384] 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)

[0385] Step A

[0386] 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] + .

[0387] Step D

[0388] Compound 22-1 (43 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 = 661.2 [M+H] + . 1 H-NMR(CDCl3)δ1.79–2.31(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).

[0389] Example 44

[0390] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 22A)

[0391] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 22B)

[0392] Compound 22 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0393] Example 45

[0394] 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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 23)

[0395] Step A

[0396] In an ice-water bath, ((S)-2-(difluoromethylene)tetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (14.28 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 23-1. LC / MS (ESI): m / z = 1036.4 [M+H] + .

[0397] Step B

[0398] Compound 23-1 (44 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 23. LC / MS (ESI): m / z = 697.2 [M+H] + . 1H-NMR(CDCl3)δ1.73-2.03(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).

[0399] Example 46

[0400] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 23A)

[0401] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 23B)

[0402] Compound 23 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0403] Example 47

[0404] 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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 24)

[0405] Step A

[0406] In an ice-water bath, compound ((S)-2-(fluoromethylene)tetrahydro-1H-pyrrolizine-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, 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 1-3. LC / MS (ESI): m / z = 1019.1 [M+H] + .

[0407] Step D

[0408] Compound 1-3 (43 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 = 679.2 [M+H] + . 1 H-NMR(CDCl3)δ1.71-2.20(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),4.30-4.98(m,7H),6.46-6.71(m,1H),6.84(d,1H).

[0409] Example 48

[0410] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 24A)

[0411] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 24B)

[0412] Compound 24 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0413] Example 49

[0414] 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)

[0415] Compound 25 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 664.2 [M+H] + .

[0416] Example 50

[0417] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 25A)

[0418] (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)

[0419] Compounds 25A and 25B were prepared using a similar method to that of compounds 2A and 2B in Example 2 (the starting material was replaced with compound 25). LC / MS (ESI): m / z = 664.2 [M+H] + .

[0420] Example 51

[0421] 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)

[0422] Compound 5 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 682.2 [M+H] + .

[0423] Example 52

[0424] (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)

[0425] (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)

[0426] Compounds 26A and 26B were prepared using a similar method to that of compounds 2A and 2B in Example 2 (the starting material was replaced with compound 26). LC / MS (ESI): m / z = 682.2 [M+H] + .

[0427] Example 53

[0428] 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)

[0429] Compound 5 was obtained by a similar preparation method to compound 1 in Example 1. LC / MS (ESI): m / z = 700.2 [M+H] + .

[0430] Example 54

[0431] (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-pyrrolazin-7a(5H)-yl)methoxy)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-6,6-dioxide (Compound 27A)

[0432] (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)

[0433] Compounds 27A and 27B were prepared using a similar method to that of compounds 2A and 2B in Example 2 (the starting material was replaced with compound 27). LC / MS (ESI): m / z = 700.2 [M+H] + .

[0434] Example 55

[0435] 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)

[0436] Compound 28 was obtained by a similar preparation method to that of compound 1 in Example 1. LC / MS (ESI): m / z = 670.2 [M+H] + .

[0437] Example 56

[0438] (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)

[0439] (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)

[0440] Compounds 28A and 28B were prepared using a similar method to that of compounds 2A and 2B in Example 2 (the starting material was replaced with compound 28). LC / MS (ESI): m / z = 670.2 [M+H] + .

[0441] Example 57

[0442] 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)

[0443] Step A

[0444] Compound F1 (82 mg, 105.02 μmol) and 8-tert-butyloxycarbonyl-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. The reaction mixture was heated to 100°C and stirred for 1 hour. After cooling, the organic solvent was removed under reduced pressure. The crude product was separated and purified by preparative thin-layer chromatography to obtain compound 29-1. LC / MS (ESI): m / z = 846 [M+H] + .

[0445] Step B

[0446] Compound 29-1 (71 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperbenzoic 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 29-2. LC / MS (ESI): m / z = 910 [M+H] + .

[0447] Step C

[0448] In an ice-water bath, compound ((2R,7AS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (12.09 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 29-2 (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 29-3. LC / MS (ESI): m / z = 989 [M+H] + .

[0449] Step D

[0450] Compound 29-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 29. LC / MS (ESI): m / z = 648.2 [M+H] + . 1H-NMR(DMSO-d6)δ1.78-2.39(m,12H),2.38-2.79(m,3H),3.13-3.44(m,3H),3 .56-3.62(m,2H),3.71-4.78(m,11H),5.92(s,2H),6.69(d,1H),6.88(d,1H).

[0451] Example 58

[0452] (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)

[0453] (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)

[0454] Compound 29 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0455] Example 59

[0456] 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)

[0457] Compound 30 was obtained by a similar preparation method to compound 29 in Example 57. LC / MS (ESI): m / z = 642.3 [M+H] + .

[0458] Example 60

[0459] (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)

[0460] (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)

[0461] Compound 30 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0462] Example 61

[0463] 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)

[0464] Compound 31 was obtained by a similar preparation method to compound 29 in Example 57. LC / MS (ESI): m / z = 679.2 [M+H] + . 1H-NMR(DMSO-d6)δ1.99(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).

[0465] Example 62

[0466] (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)

[0467] (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)

[0468] Compound 31 was separated and purified by supercritical fluid chromatography (SFC) to give 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 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 679.2 [M+H] + ).

[0469] Example 63

[0470] 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)

[0471] Compound 32 was obtained by a similar preparation method to that of compound 29 in Example 57. LC / MS (ESI): m / z = 661.2 [M+H] + . 1 H-NMR(DMSO-d6)δ1.78-2.39(m,10H),2.38-2.79(m,3H),3.13-3.44(m,3H ),3.56-3.62(m,2H),3.71-4.78(m,11H),5.92(s,2H),6.59-6.87(m,3H).

[0472] Example 64

[0473] (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)

[0474] (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)

[0475] Compound 32 was separated and purified by supercritical fluid chromatography (SFC) to give 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 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 661.2 [M+H] + ).

[0476] Example 65

[0477] 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)

[0478] Step A

[0479] Compound G1 (82 mg, 105.02 μmol) and 8-tert-butyloxycarbonyl-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. The reaction mixture was heated to 100°C and stirred for 1 hour. After cooling, the organic solvent was removed under reduced pressure. The crude product was separated and purified by preparative thin-layer chromatography to obtain compound 33-1. LC / MS (ESI): m / z = 842 [M+H]. + .

[0480] Step B

[0481] Compound 33-1 (71 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperbenzoic 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 33-2. LC / MS (ESI): m / z = 906 [M+H] + .

[0482] Step C

[0483] In an ice-water bath, compound ((2R,7AS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (12.09 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 33-2 (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 33-3. LC / MS (ESI): m / z = 985.1 [M+H] + .

[0484] Step D

[0485] Compound 33-3 (42 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 = 645.2 [M+H] + . 1 H-NMR(DMSO-d6)δ1.78-2.39(m,12H),2.38-2.79(m,3H),3.13-3.44(m,3H),3 .56-3.62(m,2H),3.71-4.78(m,11H),6.20(s,2H),6.69(d,1H),6.89(d,1H).

[0486] Example 66

[0487] (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)

[0488] (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)

[0489] Compound 33 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0490] Example 67

[0491] 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)

[0492] Compound 34 was obtained by a similar preparation method to that of compound 33 in Example 65. LC / MS (ESI): m / z = 638.2 [M+H] + .

[0493] Example 68

[0494] (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)

[0495] (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)

[0496] Compound 34 was separated and purified by supercritical fluid chromatography (SFC) to give 34A and 34B (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 = 638.2 [M+H] + ).

[0497] Example 69

[0498] 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)

[0499] Compound 35 was obtained by a similar preparation method to that of compound 33 in Example 65. LC / MS (ESI): m / z = 637.2 [M+H] + . 1 H-NMR(DMSO-d6)δ1.67-2.39(m,10H),2.38-2.79(m,3H),3.13-3.44(m,3H),3.56-3.62 (m,2H),3.71-4.78(m,11H),55.78(s,2H),6.43(s,1H),6.74(s,1H),6.76-7.19(m,1H).

[0500] Example 70

[0501] (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)

[0502] (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)

[0503] Compound 35 was separated and purified by supercritical fluid chromatography (SFC) to give 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]+ ).

[0504] Example 71

[0505] 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)

[0506] Compound 36 was obtained by a similar preparation method to that of compound 33 in Example 65. LC / MS (ESI): m / z = 619.2 [M+H] + . 1 H-NMR(DMSO-d6)δ1.60-2.39(m,9H),2.38-2.79(m,3H),3.13-3.44(m,3H),3 .56-3.62(m,2H),3.71-4.78(m,11H),6.31(s,2H),6.74(d,1H),6.92(d,1H).

[0507] Example 72

[0508] (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)

[0509] (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)

[0510] Compound 36 was separated and purified by supercritical fluid chromatography (SFC) to give 36A and 36B (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] + ).

[0511] Example 73

[0512] 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)

[0513] Step A

[0514] Compound H1 (80 mg, 105.02 μmol) and 8-tert-butyloxycarbonyl-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. The reaction mixture was heated to 100°C and stirred for 1 hour. After cooling, the organic solvent was removed under reduced pressure. The crude product was separated and purified by preparative thin-layer chromatography to obtain compound 37-1. LC / MS (ESI): m / z = 822 [M+H] + .

[0515] Step B

[0516] Compound 37-1 (70 mg, 84.96 μmol) was dissolved in dichloromethane (2 mL), and m-chloroperbenzoic 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] + .

[0517] Step C

[0518] In an ice-water bath, compound ((2R,7AS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (12.09 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 37-2 (52 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 37-3. LC / MS (ESI): m / z = 965.1 [M+H] + .

[0519] Step D

[0520] Compound 37-2 (41 mg, 42.78 μmol) was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. 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 37. LC / MS (ESI): m / z = 625.3 [M+H] + . 1 H-NMR(DMSO-d6)δ1.78-2.39(m,12H),2.38-2.79(m,3H),3.13-3.44(m,3H),3 .56-3.62(m,2H),3.71-4.86(m,11H),5.77(s,2H),6.41(s,1H),6.74(s,1H).

[0521] Example 74

[0522] (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)

[0523] (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)

[0524] Compound 37 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0525] Example 75

[0526] 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)

[0527] Compound 38 was obtained by a similar preparation method to that of compound 37 in Example 73. LC / MS (ESI): m / z = 619.2 [M+H] + .

[0528] Example 76

[0529] (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)

[0530] (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)

[0531] Compound 38 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0532] Example 77

[0533] 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)

[0534] Compound 39 was obtained by a similar preparation method to compound 37 in Example 73. LC / MS (ESI): m / z = 637.2 [M+H] + . 1 H-NMR(DMSO-d6)δ1.60-2.39(m,10H),2.38-2.79(m,3H),3.13-3.44(m,3H),3.5 6-3.62(m,2H),3.71-4.86(m,11H),5.76(s,2H),6.41(s,1H),6.58-6.88(m,2H).

[0535] Example 78

[0536] (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)

[0537] (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)

[0538] Compound 39 was separated and purified by supercritical fluid chromatography (SFC) to give 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 to -50%, 3 min; flow rate: 3.4 mL / min; wavelength: 220 nm; pressure: 1800, LC / MS (ESI): m / z = 637.2 [M+H] + ).

[0539] Example 79

[0540] 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)

[0541] Compound 40 was obtained by a similar preparation method to compound 37 in Example 73. LC / MS (ESI): m / z == 655.2 [M+H] + . 1 H-NMR(DMSO-d6)δ1.66-2.39(m,10H),2.38-2.79(m,3H),3.13-3.44(m,3H),3 .56-3.62(m,2H),3.71-4.88(m,11H),5.76(s,2H),6.41(s,1H),6.74(s,1H).

[0542] Example 80

[0543] (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)

[0544] (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)

[0545] Compound 40 was separated and purified by supercritical fluid chromatography (SFC) to give 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] + ).

[0546] Example 81 Biological Activity Test

[0547] Experimental Example 1. KRAS Inhibitory Activity Test

[0548] 1. Experimental purpose:

[0549] Through the TR-FRET method, the KRas G12D Compounds that bind to GTP.

[0550] 2. Reagent Preparation:

[0551] a. Storage of reagents:

[0552] 1) KRAS nucleotide exchange buffer

[0553] Prepare 1 L of solution by adding 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. Sterilize by filtration and store at 4°C.

[0554] 2) KRAS assay buffer

[0555] Prepare 1 L of solution by adding 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. Sterilize by filtration and store at 4°C.

[0556] 3) KRAS / Bodipy GDP / Tb-SA mixture

[0557] Take 9.5 μL of 95 μM KRas G12D Protein was mixed with 440.5 μL KRAS nucleotide exchange buffer, incubated at room temperature for 1 hour, and then mixed with 8.4 μL 17.9 μM Tb-SA, 1.8 μL 5 mM Bodipy GDP, and 9539.8 μL KRAS assay buffer to make a 1 L solution. After mixing, it was allowed to stand at room temperature for 6 hours and stored at -80 °C.

[0558] b. Experimental reagents:

[0559] 1) KRAS enzyme solution

[0560] Take 73.3 μL of KRAS / Bodipy GDP / Tb-SA mixture and 2126.7 μL of KRAS assay buffer to prepare a 2200 μL solution.

[0561] 2) SOS / GTP mixture

[0562] c. Experimental process:

[0563] Prepare a 2200 μL solution by adding 1.59 μL of 166 μM SOS protein, 198 μL of 100 mM GTP, and 2000.41 μL of KRAS assay buffer. The control compound stock solution concentration is 1 mM, and the test compound stock solution concentration is 10 mM. Transfer 9 μL of control compound and test compound to a 384-LDV plate; use Bravo to dilute the compound on the LDV plate 3-fold in 10 points; use ECHO to transfer 9 nL of the compound on the LDV plate to the experimental plate; use a Dragonfly automatic sampler to add 3 μL of 3nM Kras / 0.5nM TB-SA / 30nM BodipyGDP mixture and 3 μL Ras buffer to each well of the experimental plate in sequence, and 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 sampler to add 3 μL of 120nM SOS / 9mM GTP mixture to each well of the experimental plate, and 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 the plate and record the data; use Excel and Xlfit to analyze the data and calculate the IC of the test compound. 50 . “++++” means IC 50 ≤5nM; “+++” means 5nM <IC 50 ≤50nM; “++” means 50nM <IC 50 ≤2000nM; “+” indicates 2000nM <IC 50 .

[0564] Table 1 IC50 values ​​of compounds for KRAS enzyme inhibition.

[0565] Experimental Example 2. Cell p-ERK inhibition test

[0566] Using the HTRF method, compounds that can effectively inhibit p-ERK in AsPC-1 (G12D), A549 (G12S), HCT116 (G13D), NCI-H358 (G12C), NCI-H460 (Q61H), NCI-H727 (G12V), MKN1 (WTdep), and PSN-1 (G12R) cells were screened out.

[0567] Cells were seeded in a transparent 96-well cell culture plate, with 80 μL of cell suspension per well, each well containing 8,000 cells. The cell plate was placed in a carbon dioxide incubator and incubated overnight at 37 degrees. 2 μL of compound was added to 78 μL of cell culture medium, mixed, and 20 μL of compound solution was added to the corresponding cell plate well. The cell plate was returned to the carbon dioxide incubator and incubated for 1 hour. After the incubation, the cell supernatant was discarded and 50 μL of 1X cell lysis buffer was added to each well, and incubated at room temperature for 30 minutes. Phospho-ERK1 / 2Eu Cryptate antibody and Phospho-ERK1 / 2d2 antibody were diluted 20 times using detection buffer. 16 μL of cell lysate supernatant was taken from each well to a new 384 white microplate, and 2 μL of Phospho-ERK1 / 2Eu Cryptate antibody dilution and 2 μL of Phospho-ERK1 / 2d2 were added. Incubate with antibody diluent at room temperature for at least 4 hours. After the incubation, read the HTRF excitation at 320 nm and the emission at 615 nm and 665 nm using a multi-label analyzer.

[0568] Calculate the IC of the test compound 50 . “++++” means IC 50 ≤10nM; “+++” means 10nM <IC 50 ≤100nM; “++” means 100nM <IC 50 ≤2000nM; “+” indicates 2000nM <IC 50 .

[0569] Table 2. IC of compounds against p-ERK inhibition in tumor cells 50 (nM).

[0570] Although the present invention has been described in detail above, it will be appreciated by those skilled in the art that various modifications and variations may be made to the present invention without departing from the spirit and scope of the present invention. The scope of the present invention is not limited to the detailed description above, but rather to the claims.

Claims

1. A compound represented by general formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, Each L1 is independently selected at each occurrence from a bond, an 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 a 5-12 membered heteroaromatic group, wherein the heteroaromatic group independently at each occurrence contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S, wherein the heteroaromatic group is optionally substituted by one or more G 1 replaced by; Each X1 is independently selected at each occurrence from N, CR4; Each R2, R3, 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 carbocyclyl, 3-10 membered heterocyclic ring, 4-10 membered heterocondensed ring, 5-12 membered spiro heterocyclic ring; 3-10 membered heterocyclic ring, 4-10 membered heterocondensed ring, 5-12 membered spiro heterocyclic ring, each occurrence of which independently contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S; each R1 is independently optionally replaced by 1, 2, 3, 4, 5 or 6 heteroatoms selected from deuterium, halogen, C 1-6 Alkyl, -C 1-6 Alkoxy, oxo, OC 1-6 Alkyl, C 3-6 The carbocyclic group and the 3-10 membered heterocyclic group may be substituted or unsubstituted; U is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spirocyclyl, 5-12 membered spiroheterocyclyl, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclyl, spiroheterocyclyl, heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclyl, fused cyclyl, fused heterocyclyl, spiroheterocyclyl, aromatic or heteroaromatic is optionally substituted by one or more G 2 replaced by; G 1 , G 2 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 heteroaromatic, -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 with one or more deuterium, cyano, halogen, C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkynyl, C 3-9 Cycloalkyl or 3-9 membered heterocyclic group, C 7-10 Aryl, 6-10 membered heteroaromatic, -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 is substituted by a substituent of R9; R5, R6, R7, R8, R9 and R 10 Each independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 Alkyl, C 3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclic group, monocyclic heteroaromatic group or phenyl group; m,i is 1 or 2.

2. [Corrected 13.02.2025 according to Rule 26] A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is represented by the general formula (II-A), (II-B), (II-C) or (II-D):

3. [Corrected 13.02.2025 according to Rule 26] A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ar is selected from:

4. [Corrected 13.02.2025 according to Rule 26] A compound according to any one of Claims 1 or a pharmaceutically acceptable salt thereof, which is selected from:

5. [Corrected 13.02.2025 according to Rule 26] A compound according to any one of Claims 4 or a pharmaceutically acceptable salt thereof, which is selected from:

6. The compound according to any one of claims 1 to 5, or its optical isomers, pharmaceutically acceptable salts, prodrugs, deuterated derivatives, hydrates, or solvates, characterized in that: The pharmaceutically acceptable salt is selected from the group consisting of potassium salt, sodium salt, magnesium salt, calcium salt, sulfate, hydrochloride, phosphate, sulfonate, or carbonate.

7. A pharmaceutical composition, characterized in that Comprising the compound according to any one of claims 1 to 6, or its optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, solvate, and a pharmaceutically acceptable carrier.

8. Use of a compound according to any one of claims 1 to 6, or an optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate or solvate thereof, characterized in that: For preparing a pharmaceutical composition for treating diseases, disorders or conditions related to the activity or expression 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, characterized in that The disease, disorder or condition is selected from the following group: 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, pharyngeal cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, skin cancer, lymphoma, gastric cancer, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, monocytic leukemia, splenomegaly, polycythemia vera, multiple myeloma, myeloma and other solid tumors and blood tumors.