Pan-kras inhibitors

Abstract

This invention provides a fused-ring aromatic compound with broad-spectrum inhibitory activity against KRAS mutations, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, as shown in formula (I), wherein the definitions of each group are detailed in the specification. Furthermore, this invention also discloses pharmaceutical compositions comprising this compound and their use in the preparation of kits for treating cancer, immune diseases, or for prognostic assessment of patients with cancer, immune diseases, etc.

Description

PAN-KRAS inhibitors Technical Field

[0001] This invention relates to compounds that inhibit multiple mutant forms of KRAS, namely PAN-KRAS inhibitors. Specifically, this invention relates to PAN-KRAS compounds, pharmaceutical compositions, preparation methods, and uses thereof. Background Technology

[0002] Kirsten rat intratumoroma 2 virus oncogene homolog (KRas) is a small GTPase and a member of the Ras family. KRas protein is inactive when bound to GDP; when extracellular growth and differentiation factors transmit signals to KRas protein, they enhance GTP binding and activate it, thereby activating KRas and downstream signaling pathways. Signaling pathways such as RAS-RAF-MEK-ERK and RAS-PI3K-AKT regulate multiple cellular processes, including cell proliferation, differentiation, and survival. KRas mutations can continuously activate downstream cellular signaling, promoting cell proliferation, migration, and anti-apoptosis, and inducing tumorigenesis.

[0003] KRAS mutations are closely related to tumor formation and development. The role of KRAS in malignant tumors was observed more than 30 years ago. Approximately 20% of all human tumors show aberrant expression of KRAS, and KRAS mutations have been detected in 25-30% of lung adenocarcinomas (Samatar and Poulikakos, Nat Rev Drug Disc 2014, 13(12): 928-942). 80% of KRAS mutations occur at codon 12, causing a single amino acid substitution, the most common of which are G12D, G12V, and G12C. KRAS G12C mutation refers to the mutation of glycine at position 12 of the protein to cysteine. The incidence of tumors with this mutation is as follows: pancreatic cancer (57%), colorectal cancer (35%), bile duct cancer (28%), small bowel cancer (17%), lung cancer (16%), endometrial cancer (15%), and ovarian cancer (14%) (Seminars in Cancer Biology. 2019 Jun 27.). KRAS G12D mutation refers to the mutation of glycine at position 12 of the protein to aspartic acid. The incidence of tumors with this mutation is as follows: pancreatic cancer (25.0%), colorectal cancer (13.3%), rectal cancer (10.1%), non-small cell lung cancer (4.1%), and small cell lung cancer (1.7%) (The AACR Project GENIE Consortium, (2017) Cancer Discovery; 7(8): 818-831. Dataset Version 4). Besides G12C and G12D, KRAS has several other mutations, such as G12V, G12A, G12R, G12S, G13D, Y96D, Q61H, and Q61K. The frequency of different KRAS mutations varies in different types of cancer cells.

[0004] KRAS has become a popular anti-cancer target in the pharmaceutical industry due to the frequent mutations found in various tumor types (MeCormick (2015) Clin cancer Res. 21(8): 1797-1801). The development of small molecule inhibitors of KRAS is generally divided into three methods: (i) competitive ligand blocking GTP binding; (ii) locking KRAS G12C in an inactive state through allosteric modulation; and (iii) disrupting KRAS with its effector proteins and guanine nucleotide exchange factors (GEFs) (such as son of sevenless (SOS), RAF, and PI3K) through protein-protein interaction inhibitors.

[0005] For decades, drug development targeting KRAS largely failed, leading to its initial perception as undruggable. However, recent breakthroughs in biology and protein structure, including comparative studies of mutant and wild-type KRAS proteins, have resulted in clinical success for small-molecule inhibitors targeting KRAS G12C. Amgen's first-in-class KRAS G12C inhibitor, AMG 510, was approved by the FDA on May 28, 2021, for the treatment of locally advanced or metastatic non-small cell lung cancer with KRAS G12C mutations. Mirati and several other biopharmaceutical companies both domestically and internationally are also developing drugs targeting KRAS, but the vast majority focus on KRAS G12C or KRAS G12D mutations. As mentioned above, besides KRAS G12C or KRAS G12D mutations, KRAS has several other mutations, such as G12V, G12A, G12R, G12S, G13D, Y96D, Q61H, and Q61K. These KRAS mutations play an important role in the formation and development of many types of cancer.

[0006] Therefore, the development of drugs targeting these KRAS mutations is urgently needed. Furthermore, with the successful launch of drugs targeting KRAS G12C, we anticipate that cancer patients receiving these treatments will develop drug resistance. To address these unmet clinical needs, the development of innovative next-generation PAN-KRAS inhibitors targeting multiple KRAS mutations and combating drug resistance mechanisms is of great significance. Summary of the Invention

[0007] In one aspect, the present invention provides a novel substituted fused-ring aromatic compound that exhibits high inhibitory activity as a broad-spectrum inhibitor of KRAS mutations (PAN-KRAS inhibitor).

[0008] In one aspect, the present invention provides a compound of formula (I), a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a solvate thereof, or a prodrug thereof:

[0009]

[0010] In the formula,

[0011] R 1 Selected from -CH3, -OCH3, -OCH(CH3)2, -Cl, or cyclopropyl;

[0012] R 2 Selected from 8- to 10-membered bicyclic aryl or heteroaryl groups;

[0013] R 3 Selected from halogens, preferably -F;

[0014] m can be 0 or 1 at will;

[0015] q and n are each independently selected from 0, 1, or 2;

[0016] R for each replacement position 7 Each is independently selected from -H, -D, halogen, -CF3, -OH, -CN, -NR a R b C 1-3 Alkoxy, C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkyl COO-, p optionally being 0, 1, or 2; preferably, the C 1-3 The alkyl group is selected from methyl, ethyl, or -CH2CH2-; preferably, the hydroxyl group is C 1-3 The alkyl group is selected from -CH2OH; preferably, the C 1-3 Alkyl COO- is selected from CH3COO-;

[0017] R 41 Selected from -C 1-3 alkoxy-3 to 9-membered heterocyclic alkyl or -C 1-3 Alkylamino-3 to 9-membered heterocyclic alkyl groups;

[0018] X 1 Selected from C, O, or S;

[0019] R 5 R 6 Each is independently selected from -H, -D, and -C. 1-3 Alkyl, -OH or -C 1-3 alkyl-OH;

[0020] Preferably, when X 1 When it is C, R 5 R 6 All are H;

[0021] Both M and W are C;

[0022] The C mentioned 1-3 Alkoxy, C 1-3 Alkyl, hydroxyl C 1-3 Alkyl-, C 1-3 Alkyl C(O)O-, C 1-3 Alkylamino, 3 to 9-membered heterocyclic alkyl, 8 to 10-membered bicyclic aryl, 8 to 10-membered bicyclic heteroaryl are optionally surrounded by 0, 1, 2 or 3 independently selected from halogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, vinyl, 1-allyl, 2-allyl, ethynyl, propynyl, trifluoromethyl, amino, hydroxyl, carboxyl, p-toluenesulfonate group -C(O)NR a R b -OCH2OCH3, C 1-3 Substituents of alkyl groups COO- and -OC(O)OCH2CH3;

[0023] The C 1-3 Alkoxy, C 1-3 Two hydrogen atoms on one of the C atoms in an alkylamino group are substituted to form a 3-5 membered cycloalkyl group;

[0024] The R a R b Each is independently H, D, C 1-3 alkyl;

[0025] The heterocyclic alkyl group has at least one heteroatom selected from N, O and S as a ring atom;

[0026] The halogen is selected from F or Cl;

[0027] When X 1 For C, R 6 When it is H, the X 1 W can be chosen to be with R 6 And the H atoms on W form unsaturated double bonds;

[0028] When n is 0 and m = q = 1, the hydrogen atoms on M and its opposite C atoms are optionally substituted to form C1-3 alkylene groups.

[0029] In another aspect, the present invention provides a compound of formula (I'), a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a solvate thereof, or a prodrug thereof:

[0030]

[0031] In the formula,

[0032] R 1 Selected from -H, C 1-6 Alkyl, C 3-6 cycloalkyl, C 1-6 Alkoxy groups, halogens;

[0033] R 2 Selected from 3- to 9-membered heterocyclic alkyl groups, 5- to 6-membered monocyclic heteroaryl groups, C 6-10 Aryl or 8- to 10-membered bicyclic heteroaryl groups;

[0034] R 3 Selected from halogens;

[0035] m can be 0 or 1 at will;

[0036] q and n are each independently selected from 0, 1, or 2;

[0037] R for each replacement position 7 Each is independently selected from -H, -D, halogen, -CF3, -OH, -CN, -NR a R b -C(O)OR a C 1-6 Alkoxy, C 1-6 Alkyl, C 2-6 Alkenyl, C2-6 ynyl, p is optionally 0, 1, 2 or 3;

[0038] R 41 Selected from -C 1-3 Alkoxy-C 3-9 cycloalkyl, -C 1-3 Alkoxy-3 to 9-membered heterocyclic alkyl, -C 1-3 Alkoxy-5 to 6-membered monocyclic heteroaryl, -C 1-3 Alkoxy-C 6-10 Aryl or -C 1-3 Alkoxy-8 to 10-membered bicyclic heteroaryl groups;

[0039] R 5 R 6 Each is independently selected from -H, -D, and -C. 1-3 Alkyl, -OH, =O, halogen, -CN, -NR a R b ;

[0040] When X 1 Selected from C, O, S atoms or C 1-2 In the case of alkoxy groups, M and W are each independently selected from C atoms;

[0041] When X 1 When M and W are selected from N atoms and n = 0, M and W are each independently selected from N atoms.

[0042] The C mentioned 1-6 Alkoxy, C 1-3 Alkoxy, C 1-2 Alkoxy, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-9 Cycloalkyl, 3- to 9-membered heterocycloalkyl, 5- to 6-membered monocyclic heteroaryl, C 6-10 The aryl group, or 8 to 10-membered bicyclic heteroaryl group, is optionally surrounded by 0, 1, 2, or 3 independently selected from halogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, vinyl, 1-allyl, 2-allyl, ethynyl, propynyl, trifluoromethyl, amino, hydroxyl, carboxyl, -C(O)NR a R bSubstituents of the substituents;

[0043] The R a R b Each is independently H, D, C 1-6 alkyl;

[0044] The heterocyclic alkyl, monocyclic heteroaryl, and bicyclic heteroaryl groups have at least one heteroatom selected from N, O, and S as a ring atom;

[0045] The halogen is selected from F or Cl.

[0046] In one implementation, the C 1-6 The alkyl group is selected from methyl, ethyl, or -CH2CH2-, and Groups not included

[0047] Another aspect of the present invention provides a compound of formula (I”), a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a solvate thereof, or a prodrug thereof:

[0048]

[0049] In the formula,

[0050] R 1 Selected from -CH3, -OCH3, -OCH(CH3)2, -Cl, or cyclopropyl;

[0051] R 2 Selected from 8- to 10-membered bicyclic heteroaryl groups;

[0052] R 3 Selected from halogens, preferably -F;

[0053] m can be 0 or 1 at will;

[0054] q and n are each independently selected from 0, 1, or 2;

[0055] R for each replacement position 7 Each is independently selected from -H, -D, halogen, -CF3, -OH, -CN, -NR a R b C 1-3 Alkoxy, C 1-3 Alkyl, hydroxyl C 1-3 Alkyl group, p is optionally 0, 1 or 2; preferably, the C 1-3 The alkyl group is selected from methyl, ethyl, or -CH2CH2-; preferably, the hydroxyl group is C 1-3 The alkyl group is selected from -CH2OH;

[0056] R 41 Selected from -C1-3 alkoxy-3 to 9-membered heterocyclic alkyl groups;

[0057] X 1 Selected from C, O, and S atoms, where X 1 When it is a C atom, R 5 R 6 All are H;

[0058] Both M and W are C atoms;

[0059] The C mentioned 1-3 Alkoxy, C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, 3- to 9-membered heterocyclic alkyl, or 8- to 10-membered bicyclic heteroaryl, optionally surrounded by 0, 1, 2, or 3 independently selected from halogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, vinyl, 1-allyl, 2-allyl, ethynyl, propynyl, trifluoromethyl, amino, hydroxyl, carboxyl, -C(O)NR a R b Substituents of the substituents;

[0060] Ra and Rb are each independently H, D, and C. 1-3 alkyl;

[0061] The heterocyclic alkyl group has at least one heteroatom selected from N, O and S as a ring atom;

[0062] The halogen is selected from F or Cl.

[0063] In some embodiments, R in the compound 1 Selected from -CH3, -OCH3, -OCH(CH3)2, -Cl or cyclopropyl.

[0064] In one implementation scheme, R 2 Selected from naphthyl or indazole, wherein the naphthyl or indazole is optionally surrounded by 0, 1, 2 or 3 independently selected from halogen, ethyl, propyl, isopropyl, ethynyl, hydroxyl, p-toluenesulfonate, -C(O)NR a R b -OCH2OCH3, C 1-3 Substitution of alkylCOO-, -OC(O)OCH2CH3 or -OC(O)-phenyl groups;

[0065] Preferably, R 2 Selected from

[0066]

[0067] In one implementation scheme, R 3 Selected from -F.

[0068] In one implementation scheme, R 41 Selected from -C 1-3 alkoxy-3 to 9-membered heterocyclic alkyl or -C 1-3 Alkylamino-3 to 9-membered heterocyclic alkyl, wherein the 3 to 9-membered heterocyclic alkyl is selected from hexahydropyrrolizinyl, azircyclopentyl or morpholinyl, wherein the hexahydropyrrolizinyl, azircyclopentyl or morpholinyl is optionally substituted by 0, 1, 2 or 3 substituents independently selected from halogen, methyl, ethyl, propyl or isopropyl.

[0069] In one embodiment, the 3- to 9-membered heterocyclic alkyl group is selected from...

[0070] In one implementation, the C 1-3 Alkoxy, C 1-3 Two hydrogen atoms on any one C atom of an alkylamino group are substituted to form a cyclopropyl or cyclobutyl group.

[0071] In one implementation scheme, R 41 Selected from:

[0072]

[0073] In one implementation, R at each replacement position 7 Each is independently selected from -H, -D, halogen, -CF3, -OH, -CN, -NR a R b C 1-3 Alkoxy, C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkyl COO-, p optionally being 0, 1, or 2; preferably, the C 1-3 The alkyl group is selected from methyl, ethyl, or -CH2CH2-; preferably, the hydroxyl group is C 1-3 The alkyl group is selected from -CH2OH; preferably, the C 1-3 Alkyl COO- is selected from CH3COO-;

[0074] X 1 Choose from C or O, where X 1 When it is C, R 5 R 6 All are H;

[0075] In one implementation, the R a R b Each is independently H, D, C 1-3 alkyl.

[0076] Preferably, the R a Rb Each can be either H or methyl.

[0077] In one embodiment, the halogen is selected from F or Cl.

[0078] In one implementation scheme, Selected from The Optionally, it is divided into 0, 1, or 2 elements selected from halogen, -CF3, -OH, -CN, and -NR. a R b C 1-3 Alkoxy, C 1-3 Alkyl, hydroxyl C 1-3 Alkyl- or C 1-3 Substituents of alkyl C(O)O- groups;

[0079] Preferably, Selected from The Optionally substituted with 0, 1 or 2 substituents selected from F, -OH, -CN, amino, methyl, hydroxymethyl or -OC(O)CH3;

[0080] Preferably, Selected from

[0081] In some embodiments, the compound, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug, wherein the compound of formula (I) is as shown in formula (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ii), (Ij), (Ik), (I-1), (Im), (In), (Io), (Ip), (Iq), (Ir), (Is), (It), (Iu), (Iv), (Iw), (Ix), (Iy), or (Iz).

[0082]

[0083]

[0084]

[0085]

[0086] Among them, R 9 Selected from H, -CH2OCH3, -C(O)N(CH3)2, -C(O)CH3, -C(O)OCH2CH3,

[0087] R 1 R 3 R 41 R 7 The definitions of m, n, q, and p are as described above.

[0088] In some embodiments, the compound, its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof, wherein the pharmaceutically acceptable salt comprises any one or combination of hydrochloride, hydrobromide, sulfate, phosphate, carbonate, formate, acetate, trifluoroacetate, propionate, methanesulfonate, lactate, benzenesulfonate, p-toluenesulfonate, succinate, maleate, fumarate, tartrate, citrate, or malate.

[0089] In a preferred embodiment, the compound of formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof, wherein the compound is selected from the group consisting of:

[0090]

[0091]

[0092]

[0093]

[0094]

[0095] Where Ts is p-toluenesulfonyl and Ac is acetyl.

[0096] In another aspect, the present invention provides a method for preparing the compound of formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof, comprising the following steps,

[0097]

[0098] (1) Compound (I-1) and compound (R) 1′H) undergoes a substitution reaction to generate compound (Ma), wherein R 1′ The group is selected from R 8 or R substituted with protecting group 8 The R 8 Groups are selected from

[0099] (2) The compound (Ma) and the compound Following a Suzuki coupling reaction, compound (Mb) is generated, wherein R... 3′ The group is selected from R 1 or R substituted with protecting group 1 ;

[0100] (3) The compound (Mb) undergoes an oxidation reaction, and then reacts with R. 41 H undergoes a substitution reaction to form a compound (Mc);

[0101] (4) The compound (Mc) and the compound Following a Suzuki coupling reaction, compound (Md) is generated, wherein R... 2′ The group is selected from R 2 or R substituted with protecting group 2 ;

[0102] (5) The compound (Md) is deprotected to generate compound (M);

[0103] The R 1 R 2 R 41 R 5 R 6 R 7 M, W, X 1 The definitions of m, n, q, and p are as described above.

[0104] In another aspect, the present invention provides a pharmaceutical composition, characterized in that the composition comprises the above-described compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a solvate thereof, a prodrug thereof, and a pharmaceutically acceptable excipient.

[0105] In another aspect, the present invention provides the use of the above-mentioned compound, its pharmaceutically acceptable salt, stereoisomer, solvate, its prodrug, and the pharmaceutical composition in the preparation of medicaments for treating cancer and immune diseases or in the preparation of kits for prognostic assessment of cancer patients.

[0106] Preferably, the pharmaceutical composition further comprises another drug for treating cancer or immune diseases;

[0107] Preferably, its use in the preparation of a treatment for diseases associated with KRAS mutations;

[0108] Preferably, the cancers include, but are not limited to, pancreatic cancer, colorectal cancer, lung cancer, bile duct cancer, endometrial cancer, and ovarian cancer.

[0109] In another aspect, the present invention provides the use of the above-described compound, its pharmaceutically acceptable salt, stereoisomer, solvate, and its prodrug in the preparation of a pharmaceutical composition for KRAS inhibitors; preferably, in the preparation of KRAS G12D, KRAS G12V, KRAS G12A, KRAS G12S, KRAS G12C, KRAS G13D, KRAS Q61H, KRAS Q61K and other KRAS mutation inhibitors.

[0110] In another aspect, the present invention provides a method for inhibiting mutant KRAS in a biological sample, comprising contacting the biological sample with the compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or the pharmaceutical composition thereof.

[0111] In another aspect, the present invention provides a method for treating KRAS mutation-mediated diseases, comprising the step of administering the compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or the pharmaceutical composition thereof to a patient in need.

[0112] Preferably, the amount of the compound, its pharmaceutically acceptable salt, stereoisomer, solvate, its prodrug, or the pharmaceutical composition administered is an effective amount. Detailed Implementation

[0113] Based on the content of this disclosure, and in accordance with common technical knowledge and practices in the art, various other modifications, substitutions, or alterations can be made without departing from the basic technical ideas described in this disclosure.

[0114] I. Definition

[0115] Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "comprises" shall be understood to include the stated elements or components without excluding other elements or other components.

[0116] The compounds disclosed herein may be asymmetric, for example, having one or more stereoisomers. Unless otherwise stated, all stereoisomers are included, such as enantiomers and diastereomers. The compounds containing asymmetric carbon atoms of this disclosure can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from racemic mixtures or synthesized using chiral starting materials or chiral reagents. Racemic, diastereomer, and enantiomers are all included within the scope of this disclosure.

[0117] The disclosed compounds also include tautomer forms. The tautomer forms arise from the exchange of a single bond with an adjacent double bond, accompanied by the migration of a proton.

[0118] The term “optional” or “optionally” means that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0119] The range of numbers in this article refers to the integers within a given range. For example, "C 1-6 "" means that the group can have 1, 2, 3, 4, 5, or 6 carbon atoms; "C 3-6 "" means that the group can have 3, 4, 5 or 6 carbon atoms.

[0120] The terms "substituted" or "substituted" refer to the substitution of one or more hydrogen atoms on a particular atom or group by a substituent, provided that the valence state of the particular atom or group is normal and the resulting compound is stable. When the substituent is a ketone group (i.e., =O), it means that two hydrogen atoms are substituted. Unless otherwise specified, the type and number of substituents can be arbitrary on a chemically feasible basis.

[0121] In this disclosure, when any variable (e.g., R) n When a substituent appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted by 1-5 Rs, the group can optionally be substituted by up to 5 Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0122] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight-chain or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples 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, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-ethylpropyl, 2,3-dimethylpropyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-ethylpropyl, 2,3-ethylpropyl, 2,3-ethylpropyl, 2-methylpropyl, 2-ethylpropyl, 2-methylpropyl, 2-propyl ... -Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 2,2-diethylhexyl, 2,2-diethylhexyl, and their various branched isomers, etc. More preferably, lower alkyl groups containing 1 to 6 carbon atoms are used. Non-limiting examples 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, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group can be substituted or unsubstituted. When substituted, the substituent can be substituted at any usable connection point. The substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group. The present disclosure preferably includes methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuteralkyl, alkoxy-substituted alkyl, and hydroxy-substituted alkyl.

[0123] The term "alkenyl" refers to an alkyl group as defined above, consisting of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, etc. Alkenyl groups can be substituted or unsubstituted; when substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0124] The term "alkynyl" refers to (CH=C-), wherein the alkynyl group may be further substituted with other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, cyano, nitro, phenolic, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylic acid ester group.

[0125] The term "cycloalkyl" refers to a saturated monocyclic alkane substituent, wherein the cycloalkyl ring contains at least 3 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0126] The term "heterocyclic group" or "heterocyclic alkyl group" refers to a saturated monocyclic cyclic hydrocarbon substituent in which one or more ring atoms are selected from nitrogen, oxygen, or S(O). m (where m is an integer from 0 to 2) heteroatoms, but excluding the ring portions of -OO-, -OS-, or -SS-, with the remaining ring atoms being carbon. Non-limiting examples of heterocyclic groups include pyrroleyl, imidazolyl, tetrahydrofuranyl, tetrahydrothiopheneyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrroleyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, etc., preferably pyrroleyl, morpholinyl, piperidinyl, cycloheptyl, 1,4-diazacycloheptyl, and piperazinyl.

[0127] The heterocyclic group can be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, cyano, nitro, chloro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group.

[0128] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably 6- to 12-membered, such as phenyl and naphthyl. More preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, including benzo5- to 10-membered heteroaryl, benzo3- to 8-membered cycloalkyl, and benzo3- to 8-membered heteroalkyl, preferably benzo5- to 6-membered heteroaryl, benzo3- to 6-membered cycloalkyl, and benzo3- to 6-membered heteroalkyl, wherein the heterocyclic group is a heterocyclic group containing 1-3 nitrogen, oxygen, and sulfur atoms; or may further include a ternary nitrogen-containing fused ring containing a benzene ring.

[0129] The aryl group can be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylic acid ester group.

[0130] The term "heteroaryl" refers to a heteroaryl system containing a heteroatom and a carbon atom, wherein the heteroatom is selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5- or 6-membered, such as imidazolyl, furanyl, thiophene, thiazolyl, pyrazolyl, pyrroleyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably triazolyl, thiophene, imidazolyl, pyrazolyl, oxazolyl, pyrimidinyl, or thiazolyl; more preferably pyrazolyl, pyrroleyl, and oxazolyl.

[0131] The heteroaryl group can be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylic acid ester group.

[0132] The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, chloro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylic acid ester group.

[0133] n-BuLi: n-Butyllithium

[0134] THF: Tetrahydrofuran

[0135] DMF: N,N-dimethylformamide

[0136] MeCN: Acetonitrile

[0137] (Boc)₂O: Ditert-butyl dicarbonate

[0138] DMAP: 4-Dimethylaminopyridine

[0139] 1,4-dioxane: 1,4-dioxane

[0140] NH4SCN: Ammonium thiocyanate

[0141] acetone: Acetone

[0142] DIEA: N,N-Diisopropylethylamine

[0143] Pd(dppf)Cl2: [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride

[0144] ACN: Acetonitrile

[0145] m-CPBA: m-chloroperoxybenzoic acid

[0146] DCM: Dichloromethane

[0147] Pd(Ph3P)4: Tetra(triphenylphosphine)palladium

[0148] ((2-Fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)

[0149] Triisopropylsilane

[0150] TsOH: p-Toluenesulfonic acid

[0151] All hydrogen atoms described in this disclosure can be replaced by their isotope deuterium.

[0152] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort.

[0153] In this disclosure, It refers to the junction of chemical bonds.

[0154] In this disclosure, It refers to any existing double or single bond.

[0155] Mode middle, Represents (R) 7 p can be arbitrarily replaced Hydrogen atoms at any position on the ring, of which p R 7 They can be the same or different.

[0156] Drugs or drug compositions

[0157] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are suitable for use in human and animal tissues to the extent of reasonable medical judgment without excessive toxicity, irritation, allergic reactions, or other problems or complications in proportion to a reasonable benefit / risk ratio.

[0158] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological potency of a particular compound as a free acid or base without any adverse biological effects. Examples include acid (including organic and inorganic acids) addition salts or base addition salts (including organic and inorganic bases).

[0159] The pharmaceutically acceptable salts disclosed herein can be synthesized from parent compounds containing an acid radical or a base using conventional chemical methods. Generally, such salts are prepared by reacting these compounds, in their free acid or base form, with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0160] The pharmaceutical products or pharmaceutical compositions disclosed herein can be administered orally, topically, parenterally, or via mucosal routes (e.g., sublingually, by inhalation, or rectally) in dosage units comprising a conventional, non-toxic, pharmaceutically acceptable carrier. Oral administration is generally preferred. The active agent can be administered orally in capsule, tablet, or other similar forms (see Remington: The Science and Practice of Pharmacy, 20th Edition).

[0161] For oral administration in tablet or capsule form, the active pharmaceutical ingredient may be combined with non-toxic, pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, sucrose, glucose, mannitol, sorbitol, and other reducing and non-reducing sugars, microcrystalline cellulose, calcium sulfate, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica, stearic acid, sodium stearyl fumarate, glyceryl docosanoate, calcium stearate, etc.); disintegrants (e.g., potato starch or sodium hydroxyacetic acid starch); or wetting agents (e.g., sodium lauryl sulfate), colorants and flavorings, gelatin, sweeteners, natural and synthetic gums (e.g., gum arabic, tragacanth, or alginate), buffer salts, carboxymethyl cellulose, polyethylene glycol, waxes, etc. For oral administration in liquid form, the pharmaceutical component may be combined with a non-toxic, pharmaceutically acceptable inert carrier (e.g., ethanol, glycerol, water), an anti-settling agent (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats), an emulsifier (e.g., lecithin or gum arabic), a non-aqueous carrier (e.g., almond oil, esters, ethanol, or fractionated vegetable oils), and a preservative (e.g., methylparaben, propylparaben, or sorbic acid). Stabilizers such as antioxidants (BHA, BHT, propyl iodide, sodium ascorbate, citric acid) may also be added to stabilize the dosage form.

[0162] Tablets containing the active compound can be coated using methods well known in the art. The compositions of this disclosure containing a compound of formula I as the active compound can also incorporate beads, microspheres, or microcapsules, for example, constructed from polyglycolic acid / lactic acid (PGLA). Liquid formulations for oral administration can take the form of, for example, solutions, syrups, emulsions, or suspensions, or they can be presented as dry products reconstituted with water or other suitable excipients prior to use. Formulations for oral administration can be suitably formulated to allow for controlled or delayed release of the active compound.

[0163] The pharmaceutical products or pharmaceutical compositions disclosed herein can be delivered parenterally, i.e., administered intravenously (IV), intraventricularly (ICV), subcutaneously (SC), intraperitoneally (IP), intramuscularly (IM), subcutaneously (SD), or intradermally (ID), by direct injection, such as rapid concentration or continuous infusion. Formulations for injection may be presented in unit dosage forms, such as in ampoules or multi-dose containers with added preservatives. The compositions may be in the form of excipients, suspensions, solutions, or emulsions in oil or aqueous carriers, and may contain formulation agents such as anti-settling agents, stabilizers, and / or dispersants. Alternatively, the active ingredient may be reconstituted in powder form with a suitable carrier (e.g., sterile, pyrogen-free water) prior to use.

[0164] The pharmaceutical or pharmaceutical composition disclosed herein can also be formulated for rectal administration, for example as a suppository or retention enema (e.g., containing a conventional suppository base such as cocoa butter or other glycerides).

[0165] The term "treatment" includes suppressing, alleviating, preventing, or eliminating one or more symptoms or side effects associated with the disease, condition, or disorder being treated.

[0166] The terms “reduction,” “inhibition,” “mitigation,” or “reduction” are used relative to a control. Those skilled in the art will readily determine the appropriate control for each experiment. For example, a reduced response in a subject or cell treated with the compound is compared to a response in a subject or cell not treated with the compound.

[0167] The term "effective dose" or "therapeutic effective dose" refers to a dose sufficient to treat, suppress, or alleviate one or more symptoms of the treated disease state or otherwise provide the desired pharmacological and / or physiological effect. The precise dose will vary depending on a variety of factors, such as subject-dependent variables (e.g., age, immune system health, etc.), the disease or illness, and the treatment administered. The effect of an effective dose can be relative to a control. These controls are known in the art and discussed herein, and can be, for example, the condition of the subject before or without administration of the drug or combination of drugs, or, in the case of a combination of drugs, the combined effect can be compared to the effect of administration of only one drug.

[0168] The term "excipient" is used herein to include any other compound that is not therapeutic or biologically active and may be contained in or on microparticles. Therefore, excipients should be pharmaceutically or biologically acceptable or relevant, for example, excipients that are generally non-toxic to the subject. "Excipient" includes a single such compound and is also intended to include multiple compounds.

[0169] The term "pharmaceutical composition" means a composition comprising the compounds described in this disclosure or their pharmaceutically acceptable salts, and at least one pharmaceutically acceptable ingredient selected from the following, depending on the manner of administration and the nature of the dosage form: carriers, diluents, adjuvants, excipients, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, dispersants, thermosensitive materials, temperature regulators, adhesives, stabilizers, suspending agents, etc.

[0170] Uses and treatments

[0171] The terms “patient,” “subject,” “individual,” etc., are used interchangeably herein and refer to any animal or its cells, whether in vitro or in situ, that conform to the methods described herein. In some non-limiting embodiments, the patient, subject, or individual is a person.

[0172] According to the methods disclosed herein, compounds or compositions may be administered in any amount and via any route of administration that are effective in treating KRAS-related diseases or reducing their severity.

[0173] This disclosure relates to a method for inhibiting KRAS in a biological sample, comprising the step of contacting the biological sample with a compound of this disclosure or a composition containing the compound.

[0174] The term "biological sample" includes (but is not limited to) cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other bodily fluids or extracts thereof. Inhibition of enzymes in biological samples can be used to achieve a variety of purposes known to those skilled in the art. Examples of such purposes include (but are not limited to) bioanalysis, gene expression studies, and identification of biological targets.

[0175] The present disclosure discloses a method for inhibiting KRAS in a patient, comprising the step of administering the disclosed compound or a composition comprising the compound to the patient.

[0176] The provided compounds are KRAS inhibitors and therefore can be used to treat one or more conditions associated with KRAS activity. Therefore, in some embodiments, this disclosure provides a method for treating KRAS-mediated conditions, comprising the step of administering the disclosed compounds or pharmaceutically acceptable combinations thereof to a patient in need.

[0177] As used herein, the term "KRAS-mediated" refers to any disease, ailment, and / or symptom where KRAS or its mutants are known to act. Therefore, another embodiment of this disclosure relates to treating or reducing the severity of one or more diseases for which KRAS or its mutants are known to act.

[0178] This disclosure provides a method for treating one or more conditions, diseases, and / or symptoms, wherein the condition, disease, or symptom is a proliferative disease, such as cancer, inflammatory conditions, or viral infections.

[0179] In some embodiments, this disclosure provides a method of treating cancer or another proliferative condition, comprising administering a compound or composition of the disclosed invention to a patient suffering from cancer or another proliferative condition. In some embodiments, the method of treating cancer or another proliferative condition comprises administering a compound or composition of the disclosed invention to a mammal. In some embodiments, the mammal is a human.

[0180] As used herein, the terms “cancer inhibition” and “cancer cell proliferation inhibition” refer to inhibiting the growth, division, maturation, or survival of cancer cells, and / or causing cancer cell death through cytotoxicity, nutrient depletion, or induction of apoptosis, individually or collectively with other cancer cells.

[0181] Examples of tissues containing cancer cells whose proliferation is inhibited by the compounds and compositions described herein and to which the methods described herein are applicable include (but are not limited to) the breast, prostate, brain, blood, bone marrow, liver, pancreas, epidermis, kidney, colon, ovary, lung, testis, penis, thyroid gland, parathyroid gland, pituitary gland, thymus, retina, uvea, conjunctiva, spleen, head, neck, trachea, gallbladder, rectum, salivary glands, adrenal glands, pharynx, esophagus, lymph nodes, sweat glands, sebaceous glands, muscles, heart, and stomach.

[0182] Cancers treated with the compounds or compositions disclosed herein include, but are not limited to, melanoma, liposarcoma, lung cancer, breast cancer, prostate cancer, leukemia, kidney cancer, esophageal cancer, brain cancer, lymphoma, or colorectal cancer. In some embodiments, the cancer is primary exudative lymphoma (PEL).

[0183] The compounds disclosed herein may be used to treat proliferative diseases selected from the following: benign or malignant tumors or carcinomas of the brain, kidneys, liver, adrenal glands, bladder, breast, stomach, gastric tumors, ovaries, colon, rectum, prostate, pancreas, lungs, vagina, cervix, testes, genitourinary tract, esophagus, larynx, skin, bone, or thyroid gland; sarcomas, glioblastomas, neuroblastomas, multiple myeloma, or gastrointestinal cancers (especially colorectal cancer or colorectal adenomas) or tumors of the neck and head; epidermal hyperplasia; psoriasis; benign prostatic hyperplasia; tumor formation; epithelial-characteristic tumor formation; adenoma; adenocarcinoma; keratoacanthoma; epidermoid carcinoma; large cell carcinoma; non-small cell lung cancer; Hodgkin's and non-Hodgkin's lymphomas; breast cancer; follicular carcinoma; undifferentiated tumors; papillary carcinoma; seminoma; melanoma; MYD88-driven diseases; DLBCL; ABC. DLBCL, IL-1 driven diseases, mild or indolent multiple myeloma or leukemia.

[0184] The cancers described in this disclosure include (but are not limited to) leukemias (e.g., acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphomas (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenström's macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphoendothelial sarcoma, synovoma, mesothelioma, Ewing's tumor). Tumors, leiomyosarcomas, rhabdomyosarcomas, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystic adenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma, liver tumor, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial carcinoma, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pineal tumor, angioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, and retinoblastoma.

[0185] In some specific embodiments, the cancer is glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pineal tumor, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, or retinoblastoma.

[0186] In some specific embodiments, the cancer is an acoustic neuroma, an astrocytoma (e.g., grade I - pilocytic astrocytoma, grade II - low-grade astrocytoma, grade III - pleomorphic astrocytoma, or grade IV - glioblastoma (GBM)), a chordoma, a CNS lymphoma, a craniopharyngioma, a brainstem glioma, an ependymoma, a mixed glioma, an optic glioma, a subependymal ependymoma, a medulloblastoma, a meningioma, a metastatic brain tumor, an oligodendroglioma, a pituitary tumor, a primary neuroectodermal tumor (PNET), or a schwannoma. In some embodiments, the cancer is a type more common in children than in adults, such as a brainstem glioma, a craniopharyngioma, an ependymoma, a juvenile pilocytic astrocytoma (JPA), a medulloblastoma, an optic glioma, a pineal tumor, a primary neuroectodermal tumor (PNET), or a rhabdoid tumor. In some embodiments, the patient is an adult patient. In some embodiments, the patient is a child or a pediatric patient.

[0187] In another specific embodiment, cancers include (but are not limited to): mesothelioma, hepatobiliary (liver and bile ducts), bone cancer, pancreatic cancer, skin cancer, head or neck cancer, melanoma of the skin or eye, ovarian cancer, colorectal cancer, rectal cancer, anal cancer, gastrointestinal cancer (stomach, colon, rectum, and duodenum), uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, endocrine system cancers, thyroid cancer, parathyroid cancer, etc. Adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvis cancer, non-Hodgkin's lymphoma, spinal axis tumor, brainstem glioma, pituitary adenoma, adrenocortical carcinoma, gallbladder cancer, multiple myeloma, bile duct cancer, fibrosarcoma, neuroblastoma, retinoblastoma, or one or more of the aforementioned cancers.

[0188] In some specific embodiments, the cancer is selected from hepatocellular carcinoma, ovarian cancer, ovarian epithelial carcinoma, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine serous papillary carcinoma (UPSC); prostate cancer; testicular cancer; gallbladder cancer; bile duct hepatocellular carcinoma; soft tissue and bone synovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing's sarcoma; pleomorphic thyroid carcinoma; adrenocortical adenoma; pancreatic cancer; pancreatic duct carcinoma or pancreatic cancer; gastrointestinal / stomach (GIST) cancer; lymphoma; head and neck squamous cell carcinoma (SCCHN); salivary gland cancer; glioma or brain cancer; neurofibroma-1-associated malignant peripheral nerve sheath tumor (MPNST); Waldenström's macroglobulinemia; or medulloblastoma.

[0189] The term "primary tumor" is relative to secondary tumors. A primary tumor refers to a tumor that first appears in a certain location, such as the lungs, liver, intestines, head, or skin. It can be called primary lung cancer, primary liver cancer, primary intestinal cancer, etc.

[0190] The term "inflammatory disease" includes the aforementioned autoimmune, allergic, and inflammatory conditions, such as those selected from arthritis, ankylosing spondylitis, inflammatory bowel disease, ulcerative colitis, gastritis, pancreatitis, Crohn's disease, celiac disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, rheumatic fever, gout, organ or transplant rejection, acute or chronic graft-versus-host disease, chronic allogeneic graft rejection, Bechtel's disease, uveitis, psoriasis, dermatitis, atopic dermatitis, dermatomyositis, myasthenia gravis, Graves' disease, Hashimoto's thyroiditis, Sjögren's syndrome, and blistering conditions (e.g., pemphigus vulgaris), antibody-mediated vasculitis syndromes, including ANCA-associated vasculitis, purpura, and immune complex vasculitis (stage I or II cancer or infection). The aforementioned allergic conditions may be particularly selected from contact dermatitis, celiac disease, asthma, hypersensitivity to house dust mites, pollen and related allergens, and beryllium poisoning. The respiratory conditions mentioned may be selected in particular from asthma, bronchitis, chronic obstructive pulmonary disease (COPD), cystic fibrosis, pulmonary edema, pulmonary embolism, pneumonia, pulmonary sarcoma, silicosis, pulmonary fibrosis, respiratory failure, acute respiratory distress syndrome, primary pulmonary hypertension, and emphysema.

[0191] The term "viral infection" includes, but is not limited to, retroviral infection, hepatitis virus infection, COVID-19 infection, Zika virus infection, dengue virus infection, etc.

[0192] combination therapy

[0193] This disclosure provides combination therapies using compounds as described herein in combination with other therapeutic agents. As used herein, the term "combination therapy" includes the sequential administration of these agents, i.e., each therapeutic agent is administered at different times, and the administration of these agents, or at least two agents, substantially simultaneously. The order, or substantially simultaneous administration, of each agent may be influenced by any suitable route, including but not limited to oral, intravenous, intramuscular, subcutaneous, and direct absorption via mucosal tissue. Agents may be administered via the same or different routes. For example, a first agent may be administered orally while a second agent is administered intravenously. Furthermore, selected combinations may be administered intravenously while other agents in the combination may be administered orally. Alternatively, for example, two or more agents may be administered intravenously or subcutaneously.

[0194] Example II

[0195] The present disclosure is further illustrated below with reference to embodiments. The description of specific exemplary embodiments of the present disclosure is for illustrative and explanatory purposes. These descriptions are not intended to limit the present disclosure to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the teachings of this specification. The exemplary embodiments were chosen and described in order to explain the specific principles of the present disclosure and their practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the present disclosure, as well as various different choices and variations.

[0196] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0197] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0198] Instruments and reagents:

[0199] NMR: Agilent 400MR DD2 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as solvents, and tetramethylsilane (TMS) as the internal standard. LC-MS: Agilent 1260 Infinity II-InfinityLab LC / MSD mass spectrometer. HPLC: Agilent 1260 Infinity II high-performance liquid chromatograph (Sunfire C18 5µm 150x4.6mm column).

[0200] Thin-layer chromatography silica gel plates: HSGF254 silica gel plates (Yantai Jiangyou Silica Gel Development Co., Ltd.), specifications 0.9mm-1mm. TLC silica gel plates: GF254 silica gel plates (Yucheng Chemical (Shanghai) Co., Ltd.), specifications 0.2mm-0.25mm. Column chromatography: 300-400 mesh silica gel carrier (Qingdao Hailang Silica Gel Desiccant Co., Ltd.), Flash column (Agilent Claricep Flash amorphous silica gel purification column).

[0201] Reagents: 7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one, (R)-3-methylpiperidin-3-ol, N,N-diisopropylethylamine, 3-methylazacyclobutane-3-ol, 2,4,7-trichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidinium, 4-amino-2,6-dichloropyridine, 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octanedi(tetrafluoroborate) salt, 4-dimethylaminopyridine, ditert-butyl dicarbonate, tetra(triphenylphosphine)palladium, 1,1′-di(diphenylphosphine) (2R,7aS)-2-fluorohexahydro-1H-pyrrolizin-7a-yl)methanol, (S)-(1-methylpyrrolidine-2-yl)methanol, (2-fluoro-6-(methoxymethyloxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)naphthyl)ethynyl)triisopropylsilane, 4,4-difluoropyridine, potassium carbonate, n-butyllithium, thionyl chloride, ammonium thiocyanate, iodomethane, sodium methoxide, m-chloroperoxybenzoic acid, phosphorus oxychloride, methylboronic acid, palladium on carbon, palladium acetate, aziridine-3-ol Azacyclobutane-3-ol hydrochloride, sodium hydride, (1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane, ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolin-7a(5H)-yl)methanol, (1-(morpholinomethyl)cyclopropyl)methanol, diazepam-4-ol hydrochloride, (R)-piperidin-3-ol hydrochloride, piperidin-3-ylmethanol, cesium fluoride cesium carbonate, 1,1'-bis(diphenylphosphine)ferrocene palladium dichloromethane complex, 1,4-oxacyclopropane, azacyclobutane-3-ylmethanol hydrochloride, 1,4-diaza-2-one, 1,4 -diaza-5-one, (R)-pyrrolidine-3-ol, (S)-pyrrolidine-3-ol, dimethylcarbamoyl chloride, acetic anhydride, dimethylaminopyridine, triethylamine, p-toluenesulfonyl chloride, acetyl chloride, diethyl dicarbonate, (1-methylpyrrolidine-2-yl)methanol, 3-methylazacyclobutane-3-ol, cyclopropylboronic acid, dioxane hydrochloride, etc. Other reagents and starting materials were purchased from Shanghai Bide, Leyan Reagent Company, Jiangsu Aikon Biomedical R&D Company, Anaiji Chemical Reagent Company, Shanghai Maclean Reagent Company, Saen Chemical Reagent Company, etc., or synthesized using methods known in the art.

[0202] Unless otherwise specified, all reactions in this disclosure are carried out under continuous magnetic stirring, in dry nitrogen or argon atmosphere, in dry solvent, and at temperatures in degrees Celsius.

[0203] The following are the intermediate numbers:

[0204]

[0205] Preparation of intermediate I-1: 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1)

[0206]

[0207] (1) Synthesis of intermediate I-1a: 2,6-dichloro-3-fluoro-4-pyridineamine (I-1a)

[0208] 4-Amino-2,6-dichloropyridine (52 g, 320 mmol) was added to acetonitrile (480 ml) and N,N-dimethylformamide (480 ml), followed by the slow addition of 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane di(tetrafluoroborate) salt (136 g, 360 mmol), and the reaction was carried out at 85 °C for 3 hours. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and backwashed with saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column chromatography (petroleum ether: ethyl acetate = 0%–30%) to obtain the target product 2,6-dichloro-3-fluoro-4-pyridineamine (I-1a, 51 g, yield 88.3%). ESI [M+H] + =180.9, 181.9

[0209] (2) Intermediate I-1b: Synthesis of tert-butyl (tert-butyloxycarbonyl) (2,6-dichloro-3-fluoropyridin-4-yl) carbamate (I-1b)

[0210] 2,6-Dichloro-3-fluoro-4-pyridinamide (I-1a, 41.3 g, 229.5 mmol) and 4-dimethylaminopyridine (1.3 g, 11.6 mmol) were added to tetrahydrofuran (225 ml), followed by the slow addition of di-tert-butyl dicarbonate (125.3 g, 574.5 mmol), and the reaction was carried out at 60 °C for 3 hours. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and backwashed with saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column chromatography (petroleum ether: ethyl acetate = 0%–50%) to obtain the target product, tert-butyl(tert-butyloxycarbonyl)(2,6-dichloro-3-fluoropyridin-4-yl)carbamate (I-1b, 42.1 g, yield 48.4%). ESI [M+H] + =381.2, 382.3

[0211] (3) Intermediate I-1c: Synthesis of 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinic acid tert-butyl ester (I-1c)

[0212] Diisopropylamine (24.32 g, 240.8 mmol) was added to tetrahydrofuran (200 ml), cooled to -78 °C, and n-butyllithium (150.5 ml, 240.8 mmol) was slowly added under nitrogen protection. The reaction was carried out at -78 °C for 1 hour. Tert-butyl (tert-butyloxycarbonyl) (2,6-dichloro-3-fluoropyridin-4-yl) carbamate (I-1b, 32.6 g, 86 mmol) was dissolved in tetrahydrofuran (100 ml). l) The reaction mixture was then slowly added to the reaction solution and reacted at -78°C for 2 hours. The reaction was quenched with aqueous acetic acid, extracted with ethyl acetate, and the organic phase was collected and backwashed with saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column chromatography (petroleum ether: ethyl acetate = 0%–30%) to obtain the target product, 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinic acid tert-butyl ester (I-1c, 23.1 g, yield 95.1%). ESI [M+H] + =381.2, 382.3

[0213] (4) Synthesis of intermediate I-1d: 4-amino-2,6-dichloro-5-fluoronicotinate salt (I-1d)

[0214] 4-(tert-Butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinic acid tert-butyl ester (I-1c, 22.8 g, 60 mmol) was added to dioxane (90 ml), followed by slow addition of concentrated hydrochloric acid (30 ml) to the reaction solution. The reaction was carried out at 25 °C for 16 hours. The reaction solution was directly filtered, and the filter cake was dried to obtain the target product, 4-amino-2,6-dichloro-5-fluoronicotinic acid salt (I-1d, 18.1 g). ESI[M+H] + =225.1

[0215] (5) Synthesis of intermediate I-1e: 5,7-dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one (I-1e)

[0216] 4-Amino-2,6-dichloro-5-fluoronicotinic acid salt (I-1d, 22.8 g, 60 mmol) was added to thionyl chloride (450 ml) and reacted at 50 °C for 3 hours. The reaction solution was concentrated under reduced pressure, dissolved in acetone (90 ml), and ammonium thiocyanate (15.84 g, 138 mmol) was added. The mixture was then reacted at 25 °C for 1 hour. After filtration, the filter cake was washed with acetonitrile and dried to obtain the target product 5,7-dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one (I-1e, 17.1 g, yield >64.1%). ESI[M+H + =266.1

[0217] (6) Synthesis of intermediate I-1: 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1)

[0218] 5,7-Dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one (I-1e, 15.0 g, 56.43 mmol) was added to an aqueous sodium hydroxide solution (0.1 M, 90 mL, 56.43 mmol), followed by the slow addition of iodomethane (13.3 g, 101.6 mmol). The reaction was carried out at 25 °C for 16 hours. The reaction solution was then poured into water, and the pH was adjusted to 6. A large amount of white solid precipitated out. The solid was filtered, and the filter cake was dried to obtain the target product, 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1, 10.1 g, yield 63.9%). ESI[M+H] + =280.1

[0219] Example 1: Synthesis of 4-(4-((1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol carboxylate (1)

[0220]

[0221] Step 1: Synthesis of (1R,5S)-3-(5,7-dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-a)

[0222] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 300 mg, 1.07 mmol) was dissolved in acetonitrile (10 ml), and then phosphorus oxychloride (246 mg, 1.605 mmol) and N,N-diisopropylethylamine (415 mg, 3.21 mmol) were added. After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (207 mg, 1.605 mmol) and (1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane (72 mg, 0.636 mmol) were added. The reaction was carried out at 0 °C for 0.5 hours. After concentration under reduced pressure, TLC (methanol:dichloromethane = 50:1) purification yielded the target product (1R,5S)-3-(5,7-dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-a, 300 mg, yield 74.63%). MS (ESI) [M+H] + 375.1, 377.0

[0223] Step 2: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-b)

[0224] (1R,5S)-3-(5,7-dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-a, 300 mg, 0.799 mmol) was dissolved in toluene (5 ml) and water (0.5 ml), and then methylboronic acid (191 mg, 3.196 mmol), potassium phosphate (509 mg, 2.4 mmol), and 1,1′-bis(diphenylphosphino)ferrocene palladium dichloride (II) (59 mg, 0.08 mmol) were added. The mixture was heated to 105 °C and reacted for 24 h. After concentration under reduced pressure, TLC (petroleum ether: ethyl acetate = 3:1) purified the target product (1R, 5S)-3-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-b, 125 mg, yield 44.01%). MS (ESI) [M+H] + 355.0, 357.0

[0225] Step 3: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-5-methyl-2-(methanesulfonyl)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-c)

[0226] (1R,5S)-3-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-b, 100 mg, 0.282 mmol) and m-chloroperoxybenzoic acid (146 mg, 0.846 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 1 h. The reaction was quenched with water, extracted with dichloromethane, and the organic phase was collected and washed with water in saturated sodium bicarbonate solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target product (1R,5S)-3-(7-chloro-8-fluoro-5-methyl-2-(methylsulfonyl)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-c, 130 mg). MS(ESI)[M+H] + 387.1, 389.1

[0227] Step 4: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-d)

[0228] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolino-7a(5H)-yl)methanol (160 mg, 1.008 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C and 60% sodium hydride (42 mg, 1.05 mmol) was added, and the reaction was continued for 0.5 hours. Then (1R,5S)-3-(7-chloro-8-fluoro-5-methyl-2-(methanesulfonyl)pyrimidin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-c 160 mg, 0.41 mmol), reacted for 0.5 h, concentrated under reduced pressure, and purified by TLC (dichloromethane:methanol = 10:1) to obtain the target product (1R,5S)-3-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-d, 72 mg, yield 45.86%). MS (ESI) [M+H] + 466.2, 468.2

[0229] Step 5: Synthesis of (1R,5S)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-e)

[0230] The following ingredients were added: (1R,5S)-3-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-d, 70 mg, 0.15 mmol), potassium carbonate (83 mg, 0.6 mmol), and tetraphenylphosphine. Palladium (17 mg, 0.015 mmol) and (2-fluoro-6-(methoxymethyloxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (115 mg, 0.225 mmol) were dissolved in 1,4-dioxane (3 ml) and water (0.75 ml), purged with nitrogen for 1 minute, and reacted in a microwave at 135 °C for 1 hour. After concentration under reduced pressure, TLC (dichloromethane:methanol = 15:1) yielded the target product (1R,5S)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-e, 62 mg, yield 50.82%). MS (ESI) [M+H] + 816.9, 817.8

[0231] Step 6: Synthesis of (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-f)

[0232] (1R,5S)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-e, 60 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (64 mg, 0.42 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), extracted three times with ethyl acetate (20 ml each time), and the organic phase was collected and distilled under reduced pressure to give the target product (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (1-f, 51 mg, yield 111%). MS (ESI) [M+H] + 660.7, 661.7

[0233] Step 7: Synthesis of 4-(4-((1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol carboxylate (1)

[0234] (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (1-f, 51 mg, 0.077 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (0.5 ml) was added at 25 °C. The reaction was allowed to proceed for 0.5 h. The reaction solution was purified by synthetic liquid chromatography to obtain the target product 4-(4-(((1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol carboxylate (1, 0.69 mg, yield 1.35%). MS (ESI) [M+H] +616.6, 617.6

[0235] Example 2: Synthesis of 4-(4-((1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol carboxylate (2)

[0236]

[0237] Step 1: Synthesis of 7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a)

[0238] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 300 mg, 1.07 mmol) was dissolved in N,N-dimethylacetamide (3 ml) and methanol (0.5 ml), followed by the addition of sodium methoxide (288 mg, 5.35 mmol). The reaction was carried out at 50 °C for 16 hours, then cooled to 0 °C. The pH was adjusted to 3 with concentrated hydrochloric acid, filtered, and the filter cake was washed three times with water and dried to obtain the target product, 7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 300 mg, yield 94.63%). MS (ESI) [M+H]+ 276.1, 277.0

[0239] Step 2: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-b)

[0240] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 295.3 mg, 1.07 mmol) was dissolved in acetonitrile (10 ml), and then phosphorus oxychloride (246 mg, 1.605 mmol) and N,N-diisopropylethylamine (415 mg, 3.21 mmol) were added. After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (207 mg, 1.605 mmol) and (1R,5S)-8-oxa-3-azabicyclo[3.2.1]octane (72 mg, 0.636 mmol) were added. The reaction was carried out at 0 °C for 0.5 hours. After concentration under reduced pressure, TLC (methanol:dichloromethane = 10:1) purification yielded the target product (1R,5S)-3-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-b, 300 mg, yield 74.63%). MS (ESI) [M+H] + 371.1

[0241] Step 3: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-c)

[0242] (1R,5S)-3-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-b, 300 mg, 0.564 mmol) and m-chloroperoxybenzoic acid (292 mg, 1.68 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 1 h. The solution was quenched with water, extracted with dichloromethane, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered and concentrated under reduced pressure to obtain the target product (1R,5S)-3-(7-chloro-8-fluoro-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-c, 320 mg, yield 98.5%). MS (ESI) [M+H] + 403.1, 404.1

[0243] Step 4: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (2-d)

[0244] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolino-7a(5H)-yl)methanol (160 mg, 1.008 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C and 60% sodium hydride (42 mg, 1.05 mmol) was added, and the reaction was continued for 0.5 hours. Then (1R,5s)-3-(7-chloro-8-fluoro-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-c 160 mg, 0.43 mmol), reacted for 0.5 h, concentrated under reduced pressure, and purified by TLC (dichloromethane:methanol = 10:1) to obtain the target product (1R,5S)-3-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (2-d, 72 mg, yield 37.70%). MS (ESI) [M+H] + 482.1

[0245] Step 5: Synthesis of ((1R,5S)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-e)

[0246] The following ingredients were added: (1R,5S)-3-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)-8-oxa-3-azabicyclo[3.2.1]octane (2-d, 72 mg, 0.15 mmol), potassium carbonate (83 mg, 0.6 mmol), and tetraphenylphosphine. Palladium (17 mg, 0.015 mmol) and ((2-fluoro-6-(methoxymethyloxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (115 mg, 0.225 mmol) were dissolved in 1,4-dioxane (3 ml) and water (0.75 ml), purged with nitrogen for 1 minute, and reacted in a microwave oven at 135 °C for 1 hour. After concentration under reduced pressure, TLC (dichloromethane:methanol = 15:1) yielded the target product (1R,5S)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-e, 60 mg, yield 50.82%). MS (ESI) [M+H] + 832.4

[0247] Step 6: Synthesis of (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-f)

[0248] (1R,5S)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-e, 60 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (64 mg, 0.42 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction solution was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to obtain the target product (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-f, 51 mg). MS (ESI) [M+H] + 676.1

[0249] Step 7: Synthesis of 4-(4-((1R,5S)-8-oxo-3-azabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridin[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol (2)

[0250] (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-8-oxo-3-azabicyclo[3.2.1]octane (2-f, 51 mg, 0.077 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloric acid solution (0.5 ml) was added at 25 °C. The reaction was allowed to proceed for 0.5 h. The reaction solution was purified by synthetic liquid chromatography to obtain the target product 4-(4-((1R,5S)-8-oxo-3-azabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methoxypyridin[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthyl-2-ol (2, 0.47 mg). MS (ESI) [M+H] + 632.1, 1H NMR (600MHz, DMSO-d6), δ10.17 (s, 1H), 8.17-7.90 (m, 2H), 7.44 (t, J=9.0Hz, 1H), 7.36 (d, J=2.6Hz, 1H), 7.20 (s, 1 H), 5.30 (s, 1H), 5.21 (s, 1H), 4.41 (d, J=12.0Hz, 2H), 4.10 (dd, J=10.3, 4.2Hz, 1H), 4.04-3.97 (m, 2H), 3.91 (s, 1H) , 3.87 (s, 3H), 3.42 (d, J = 13.1Hz, 3H), 3.06 (d, J = 11.4Hz, 2H), 3.01 (s, 1H), 2.81 (q, J = 8.0, 7.2Hz, 1H), 2.09 (d, J = 4.9Hz, 1H), 2.04 (s, 1H), 1.98 (dd, J=8.3, 4.2Hz, 1H), 1.83 (d, J=6.6Hz, 1H), 1.81-1.76 (m, 4H), 1.75-1.71 (m, 2H).

[0251] Example 3: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)azacycloheptan-4-ol carboxylate (3)

[0252]

[0253] Step 1: Synthesis of 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclohepta-4-ol (3-a)

[0254] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 300 mg, 1.07 mmol) was dissolved in acetonitrile (10 ml), and then phosphorus oxychloride (246 mg, 1.605 mmol) and N,N-diisopropylethylamine (415 mg, 3.21 mmol) were added. After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (207 mg, 1.605 mmol) and diazepam-4-ol hydrochloride (63 mg, 0.636 mmol) were added. The reaction was carried out at 0 °C for 0.5 hours. After concentration under reduced pressure, TLC (methanol:dichloromethane = 50:1) purified the target product 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclohepta-4-ol (3-a, 300 mg, yield 77.92%). ESI [M+H]+ =377.1

[0255] Step 2: Synthesis of 4-(azacyclohepta-1-ol)-7-chloro-8-fluoro-5-methyl-2-(methylthio)pyridine[4,3-d]pyrimidine (3-b)

[0256] 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacycloheptan-4-ol (3-a, 287 mg, 0.799 mmol) was dissolved in toluene (5 mL) and water (0.5 mL), followed by the addition of methylboronic acid (191 mg, 3.196 mmol), potassium phosphate (509 mg, 2.4 mmol), and [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride (59 mg, 0.08 mmol). The mixture was heated to 105 °C and reacted for 24 h. After concentration under reduced pressure, the product was purified by TLC (petroleum ether:ethyl acetate = 3:1) to obtain the target product 4-(azacycloheptan-1-ol)-7-chloro-8-fluoro-5-methyl-2-(methylthio)pyridin[4,3-d]pyrimidinyl (3-b, 200 mg, yield 73.8%). ESI [M+H] + =357.1

[0257] Step 3: Synthesis of 4-(azacyclohepta-1-ol)-7-chloro-8-fluoro-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidine (3-c)

[0258] 4-(azacyclohepta-1-ol)-7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidine (3-b, 200 mg, 0.578 mmol) and m-chloroperoxybenzoic acid (292 mg, 1.681 mmol) were dissolved in dichloromethane (10 mL). The reaction was carried out at room temperature for 1 h. The reaction was quenched with water, extracted with dichloromethane, and the organic phase was collected and washed with water in saturated sodium bicarbonate solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the target product 4-(azacyclohepta-1-ol)-7-chloro-8-fluoro-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidine (3-c, 218 mg, 99.1% yield). ESI[M+H] + =390.1

[0259] Step 4: Synthesis of 4-(azacycloheptan-1-ol)-7-chloro-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridine[4,3-d]pyrimidine (3-d)

[0260] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methanol (87 mg, 1.17 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0°C, and 60% sodium hydride (42 mg, 1.05 mmol) was added. After reacting for another 0.5 hours, 4-(azacycloheptan-1-ol)-7-chloro-8-fluoro-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidine The pyrimidine (3-c, 218 mg, 0.584 mmol) was reacted for 0.5 h, concentrated under reduced pressure, and purified by TLC (dichloromethane:methanol = 10:1) to give the target product 4-(azacycloheptan-1-ol)-7-chloro-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidine (3-d, 132 mg, yield 51.11%). ESI[M+H] + =469.2

[0261] Step 5: Synthesis of 4-(azacycloheptan-1-ol)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridinium[4,3-d]pyrimidine (3-e)

[0262] 4-(azacycloheptan-1-ol)-7-chloro-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidine (3-d, 132 mg, 0.29 mmol), potassium carbonate (83 mg, 0.6 mmol), tetrakis(triphenylphosphine)palladium (17 mg, 0.015 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (115 mg, 0.225 mmol) were dissolved in 1,4-dioxane (3 ml) and water (0.75 ml), purged with nitrogen for 1 minute, and microwaved at 135°C for 1 hour. After concentration under reduced pressure, TLC (dichloromethane:methanol = 15:1) yielded the target product 4-(azacycloheptan-1-ol)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin [4,3-d]pyrimidine (3-e, 117 mg, yield 50.82%). ESI [M+H] + =819.4

[0263] Step 6: Synthesis of 4-(azacyclohepta-1-ol)-7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridinium[4,3-d]pyrimidine (3-f)

[0264] 4-(azacycloheptan-1-ol)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridinium[4,3-d]pyrimidine (3-e, 117 mg, 0.15 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (128 mg, 0.84 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to give the target product 4-(azacyclohepta-1-ol)-7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin [4,3-d]pyrimidine (3-f, 91 mg, yield 96.8%). ESI [M+H] + =663.2

[0265] Step 7: 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)azacycloheptan-4-ol carboxylate (3)

[0266] 4-(azacycloheptan-1-ol)-7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridinium[4,3-d]pyrimidine (3-f, 91 mg, 0.141 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (2 ml) was added at 25 °C. The reaction was allowed to proceed for 0.5 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)azacyclohepta-4-ol carboxylate (3, 0.78 mg, yield 1.35%). ESI [M+H] + =619.3

[0267] Example 4: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-4-((3R)-3-hydroxycyclohexyl)-5-methylpyridin[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate (4)

[0268]

[0269] Step 1: Preparation of (1R)-3-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-a)

[0270] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 400 mg, 1.45 mmol) was dissolved in acetonitrile (10 ml), and then phosphorus oxychloride (1.67 g, 10.9 mmol) and N,N-diisopropylethylamine (1.69 g, 13.05 mmol) were added. After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (2.81 g, 21.75 mmol) and (R)-piperidine-3-ol hydrochloride (317 mg, 2.31 mmol) were added. The reaction was carried out at 0 °C for 0.5 hours. Preparation of the target product (1R)-3-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-a, 242 mg, yield 46.73%) after vacuum concentration and flash purification (petroleum ether:ethyl acetate = 4:1). (ESI)[M+H + =362.2

[0271] Step 2: (1R)-3-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol(4-b)

[0272] (1R)-3-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-a, 192 mg, 0.53 mmol) was dissolved in toluene (8 ml) and water (0.8 ml), and then methylboronic acid (159 mg, 2.65 mmol), potassium phosphate (450 mg, 2.12 mmol), and 1,1′-bis(diphenylphosphino)ferrocene palladium dichloride (II) (78 mg, 0.106 mmol) were added. The mixture was heated to 105 °C and reacted for 24 h. After concentration under reduced pressure, TLC (dichloromethane:methanol = 100:3) purification yielded the target product (1R)-3-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-b, 114 mg, yield 50.1%). (ESI)[M+H + =342.1

[0273] Step 3: Preparation of (1R)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilylethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-c)

[0274] (1R)-3-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-b, 77 mg, 0.224 mmol), potassium carbonate (124 mg, 0.896 mmol), tetrakis(triphenylphosphine)palladium (52 ​​mg, 0.0448 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (138 mg, 0.269 mmol) were dissolved in 1,4-dioxane (3 ml) and water (0.75 ml), purged with nitrogen for 1 minute, and microwaved at 135 °C for 1.5 hours. Preparation of the target product (1R)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilylethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol by TLC (dichloromethane:methanol = 15:1) after concentration under reduced pressure (4-c, 87 mg, yield 56.1%).

[0275] Step 4: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (4-d)

[0276] (1R)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilylethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-c, 87 mg, 0.127 mmol), and m-chloroperoxybenzoic acid (66 mg, 0.381 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 1 h. Quenching with aqueous solution, extraction with dichloromethane, collection of organic phase and washing with saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (4-d, 88 mg, yield 96.3%).

[0277] Step 5: Preparation of (1R)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-e)

[0278] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolino-7a(5H)-yl)methanol (58 mg, 0.366 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (15 mg, 0.366 mmol) was added. After reacting for another 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-] [d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (4-d), purified by dichloromethane:methanol = 10:1 to give the target product (1R)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-e, 32 mg, yield 33.1%). (ESI)[M+H + =803.1

[0279] Step 6: (1R)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol(4-f)

[0280] (1R)-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-e, 32 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (2 ml), and cesium fluoride (36 mg, 0.24 mmol) was added. The mixture was reacted at 25 °C for 0.5 hours. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to give the target product (1R)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-f, 30 mg, yield 120%). (ESI)[M+H + =647.2

[0281] Step 7: Preparation of 5-ethynyl-6-fluoro-4-(8-fluoro-2-((2R,7as)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-4-((3R)-3-hydroxycyclohexyl)-5-methylpyridin[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate (4)

[0282] (1R)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)cyclohexane-1-ol (4-f, 25 mg, 0.04 mmol) was dissolved in acetonitrile (5 ml), and at 25 °C, 1,4-dioxane hydrochloride solution (0.5 ml) was added, and the reaction was carried out for 0.5 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product 5-ethynyl-6-fluoro-4-(8-fluoro-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-4-((3R)-3-hydroxycyclohexyl)-5-methylpyridin[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate (4, 1.98 mg, yield 11.08%). (ESI)[M+H + =603.2, 1 H NMR (400MHz, DMSO-d6) δ10.17 (s, 1H), 7.97 (dd, J=9.2, 5.9Hz, 1H), 7.45 (t, J=9.0Hz, 1H), 7.39 (d, J=2.6Hz, 1H), 7.29 (d, J=2.5Hz, 1H), 5.35 (s, 1H), 5.22 (s, 1H), 4.65-4.58 (m, 2H), 4.13 (d, J=11.7Hz, 2H), 3.98 (d, J=10.4Hz, 1H), 3.60 ( d, J=13.1Hz, 1H), 3.41 (s, 1H),, 3.02 (s, 1H), 2.86-2.80 (m, 1H), 2.69-2.61 (m, 1H), 2.56 (s, 3H), 2.13 (s, 1H), 2.07-2 .05 (m, 1H), 2.00 (s, 1H), 1.86-1.81 (m, 1H), 1.80-1.75 (m, 2H), 1.67-1.62 (m, 2H), 1.50 (d, J=13.0Hz, 1H), 1.4 (s, 3H).

[0283] Example 5: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate (5)

[0284]

[0285] Step 1: Synthesis of 7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a)

[0286] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1, 743 mg, 2.65 mmol) was dissolved in methanol (0.7 mL) and N,N dimethylacetamide (4.0 mL), followed by the addition of sodium methoxide (1.15 g, 21.2 mmol). The mixture was reacted at 50 °C for 16 hours, then cooled to 0 °C. Water was added to precipitate the solid, which was then adjusted to pH 3 with concentrated hydrochloric acid. The mixture was filtered and dried to obtain the target product, 7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 787 mg, yield 43.1%). ESI[M+H] + =275.9

[0287] Step 2: Synthesis of 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-a)

[0288] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 716.8 mg, 2.6 mmol) was dissolved in acetonitrile (30 mL), followed by the addition of N,N-diisopropylethylamine (2.6 mL, 15.6 mmol) and phosphorus oxychloride (0.72 mL, 7.8 mmol). After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (1.3 mL, 3 mmol) and 3-methylpiperidin-3-ol (394.23 mg, 1 mmol) were added, followed by a reaction at 0 °C for 0.5 hours. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–40%) to obtain the target product 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-a, 430 mg, yield 26.6%). ESI [M+H] + =373.0

[0289] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-b)

[0290] 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-a, 134 mg, 0.36 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (276.8 mg, 0.54 mmol), and potassium carbonate (199 mg, 1.44 mmol) were dissolved in a mixed solvent (10 mL) of 1,4-dioxane and water (4:1), and then tetra(triphenylphosphine)palladium (41.6 mg, 0.1 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (petroleum ether: ethyl acetate = 3:1) to obtain the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-b, 171 mg).

[0291] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-c)

[0292] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-b, 171 mg, 0.23 mmol) was dissolved in dichloromethane (3 mL), and then m-chloroperoxybenzoic acid (59.5 mg, 0.345 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to obtain crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-c, 150 mg)

[0293] Step 5: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-d)

[0294] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (95.52 mg, 0.6 mmol) was dissolved in ultra-dry tetrahydrofuran (2 mL), sodium hydrogen (9.6 mg, 0.4 mmol) was added, and the mixture was stirred at 0 °C for 30 minutes. Then, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-c, 150 mg, 0.4 mmol) was added. A solution of 0.2 mmol) dissolved in tetrahydrofuran (2 mL) was added to the reaction mixture, and the reaction was carried out at room temperature for 30 min. The reaction was quenched with water, extracted with ethyl acetate, and concentrated under reduced pressure to give the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-d, 294 mg, yield 52.8%). ESI[M+H + =834.2

[0295] Step 6: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-e)

[0296] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-d, 594 mg, 0.71 mmol) was dissolved in N,N dimethylformamide (5 mL), and cesium fluoride (647 mg, 4.26 mmol) was added. The reaction was carried out at room temperature for 1 hour. The sample was extracted with ethyl acetate after adding saturated brine and concentrated under reduced pressure to give the crude product 1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-e, 816 mg, yield 33.9%). ESI[M+H + =678.2

[0297] Step 7: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate (5)

[0298] 1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5-e, 200 mg, 0.295 mmol) was dissolved in acetonitrile (5 mL), and 1,4-dioxane hydrochloride solution (0.9 mL) was added. The reaction was carried out at room temperature for 1.5 hours. The pH was adjusted to 7 with a saturated sodium bicarbonate solution. The reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate (5, 2.51 mg, yield 1.34%). ESI[M+H] + =634.2 1 H NMR(400MHz, DMSO-d6)δ 8.28 (s, 2H), 7.94 (dd, J=9.2, 5.9Hz, 1H), 7.44 (td, J=8.9, 3.9Hz, 1H), 7.38-7.32 (m, 1H), 7.22(dd, J=24.7, 2.5Hz, 1H), 5.32(s, 1H), 5.19(s, 1H), 4.58(s, 1H), 4.39(s, 1H ), 4.14-4.02(m, 2H), 4.01-3.90(m, 2H), 3.86(d, J=4.4Hz, 3H), 2.99(s, 2H), 2.80(d, J=7.3Hz, 2H), 2.03-1.98 (m, 2H), 1.84-1.76 (m, 2H), 1.66-1.48 (m, 6H), 1.21 (s, 3H).

[0299] Example 6: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)piperidin-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphthalene-2-ol formate (6)

[0300]

[0301] Step 1: Synthesis of (1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-a)

[0302] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 276 mg, 1.0 mmol) was dissolved in acetonitrile (12 mL), followed by the addition of N,N-diisopropylethylamine (1.04 mL, 6.0 mmol) and phosphorus oxychloride (0.28 mL, 3.0 mmol). After reacting at 80 °C for 1.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (0.5 mL, 3 mmol) and piperidin-3-ylmethanol (115.2 mg, 1 mmol) were added, followed by a reaction at 0 °C for 2 hours. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–40%) to obtain the target product 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-a, 82 mg, yield 22%). ESI[M+H + =373.1

[0303] Step 2: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-b)

[0304] 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-a, 82 mg, 0.22 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (169.1 mg, 0.33 mmol), and potassium carbonate (121.6 mg, 0.88 mmol) were dissolved in a mixed solvent (4 mL) of 1,4-dioxane and water (4:1), and then tetra(triphenylphosphine)palladium (25.4 mg, 0.022 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the crude product was purified by thin-layer chromatography using silica gel plates (petroleum ether: ethyl acetate = 3:1) to obtain (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-b, 78 mg)

[0305] Step 3: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-c)

[0306] (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-b, 78 mg, 0.108 mmol) was dissolved in dichloromethane (1 mL), and then m-chloroperoxybenzoic acid (28 mg, 0.162 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to obtain crude product (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-c, 91 mg)

[0307] Step 4: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-d)

[0308] Dissolve ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (57.3 mg, 0.36 mmol) in ultra-dry tetrahydrofuran (1 mL), add sodium hydrogen (6 mg, 0.24 mmol), stir at 0 °C for 30 minutes, then add a solution of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-c, 91 mg, 0.12 mmol) dissolved in tetrahydrofuran (1 mL). React at room temperature for 30 minutes. Quenching with water, extraction with ethyl acetate, and concentration under reduced pressure yielded the crude product (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-d, 21 mg, yield 12.6%). ESI[M+H + =834.1

[0309] Step 5: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-e)

[0310] (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-d, 21 mg, 0.025 mmol) was dissolved in N,N dimethylformamide (1 mL), and cesium fluoride (22.8 mg, 0.15 mmol) was added. The reaction was carried out at room temperature for 1 hour. Extracted with saturated brine and ethyl acetate, the crude product was concentrated under reduced pressure to obtain 1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-e, 15 mg, yield 26.6%). ESI[M+H + =678.4

[0311] Step 6: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5HH)-yl)methoxy)-4-(3-(hydroxymethyl)piperidin-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphthalene-2-ol formate (6)

[0312] 1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (6-e, 15 mg, 0.022 mmol) was dissolved in acetonitrile (2 mL), and 1,4-dioxane hydrochloride solution (0.4 mL) was added. The reaction was carried out at room temperature for 1 hour. The pH was adjusted to 7 with a saturated sodium bicarbonate solution. The reaction solution was purified by preparative liquid chromatography to obtain the target product, 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)piperidin-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate (6, 0.58 mg, yield 4.16%). ESI[M+H] + =634.0, 1 H NMR (600MHz, DMSO-d6) δ10.20 (s, 1H), 8.32 (s, 2H), 7.94 (dd, J=9.2, 5.9Hz, 1H), 7 .44 (t, J=9.0Hz, 1H), 7.36 (d, J=2.6Hz, 1H), 7.21 (d, J=6.2Hz, 1H), 5.30 (s, 1H), 5. 21 (s, 1H), 4.61 (s, 1H), 4.21 (s, 1H), 4.08 (td, J=11.1, 5.3Hz, 2H), 4.00 (d, J=7.8 Hz, 3H), 3.87 (s, 3H), 2.80 (d, J=8.4Hz, 2H), 2.11-1.96 (m, 4H), 1.89-1.59 (m, 9H).

[0313] Example 7: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate (7)

[0314]

[0315] 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (5.17 mg, 0.027 mmol) was dissolved in tetrahydrofuran (1 mL), and palladium on carbon (6 mg) was added. The reaction was carried out at room temperature for 4 hours under hydrogen protection. The mixture was then filtered with diatomaceous earth, the filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate (7, 2.96 mg, yield 17.2%). ESI[M+H] + =637.2. 1 H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 8.17 (s, 1H), 7.76 (td, J=6.2, 2.8Hz, 1H), 7.40-7.24 (m, 2H), 7.07 (d, J=9.2Hz, 1H), 5. 35 (s, 1H), 5.21 (s, 1H), 4.56 (s, 1H), 4.42 (s, 1H), 4.17-4.09 (m, 1H), 4.02 (dd, J=13.2, 9.2Hz, 1H), 3.98-3.83 (m, 3H), 3.57 (d, J=14.8Hz, 1H), 3.47 (d, J=31.0Hz, 1H), 3.05 (d, J=31.8Hz, 3H), 2.83 (d, J=7.6Hz, 1H), 2.29 (s, 2H), 2.13 (s, 1H), 2.03 (d , J=20.8Hz, 2H), 1.81 (d, J=36.2Hz, 4H), 1.63 (s, 2H), 1.24 (s, 1H), 1.07 (s, 1H), 1.01 (s, 1H), 0.80 (dt, J=14.6, 7.6Hz, 2H).

[0316] Example 8: Synthesis of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidine-3-ol carboxylate (8)

[0317]

[0318] Step 1: Synthesis of (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-a)

[0319] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 966 mg, 3.5 mmol) was dissolved in acetonitrile (40 mL), followed by the addition of N,N-diisopropylethylamine (3.7 mL, 21 mmol) and phosphorus oxychloride (1 mL, 10.5 mmol). After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (3.7 mL, 21 mmol) and (R)-piperidin-3-ol (482 mg, 3.5 mmol) were added, followed by a reaction at 0 °C for 3 hours. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–50%) to obtain the target product (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-a, 627 mg, yield 29.9%). ESI[M+H + =358.5

[0320] Step 2: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-b)

[0321] (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-a, 251 mg, 0.7 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphthyl)ethynyl)triisopropylsilane (538 mg, 1.05 mmol), and potassium carbonate (387 mg, 2.8 mmol) were dissolved in a mixed solvent (15 mL) of 1,4-dioxane and water (4:1), and then tetra(triphenylphosphine)palladium (81 mg, 0.07 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the crude product was purified by thin-layer chromatography using silica gel plates (petroleum ether: ethyl acetate = 3:1) to obtain (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-b, 592 mg).

[0322] Step 3: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-c)

[0323] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-b, 592 mg, 0.83 mmol) was dissolved in dichloromethane (8 mL), and then m-chloroperoxybenzoic acid (214 mg, 1.245 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, and concentrate under reduced pressure to obtain crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-c, 480 mg)

[0324] Step 4: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-d)

[0325] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (205 mg, 1.29 mmol) was dissolved in ultra-dry tetrahydrofuran (3 mL), sodium hydrogen (20 mg, 0.86 mmol) was added, and the mixture was stirred at 0 °C for 30 minutes. Then, a solution of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-c, 311 mg, 0.43 mmol) dissolved in tetrahydrofuran (3 mL) was added. The reaction was carried out at room temperature for 30 minutes. Quenching with water, extraction with ethyl acetate, and concentration under reduced pressure yielded the crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridano[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-d, 356 mg, yield 40.3%). ESI[M+H+ =820.1

[0326] Step 5: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-e)

[0327] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-d, 356 mg, 0.43 mmol) was dissolved in N,N dimethylformamide (4 mL), and cesium fluoride (391 mg, 2.58 mmol) was added. The reaction was carried out at room temperature for 1 hour. Extracted with saturated brine and ethyl acetate, the crude product was concentrated under reduced pressure to obtain (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridano[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-e, 200 mg, yield 14.1%). ESI[M+H + =663.7

[0328] Step 6: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-f)

[0329] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-e, 200 mg, 0.3 mmol) was dissolved in acetonitrile (4 mL), and 1,4-dioxane hydrochloride solution (3 mL) was added. The reaction was carried out at room temperature for 1 hour. The pH was adjusted to 7 with a saturated sodium bicarbonate solution. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-f, 32 mg, yield 172%). ESI[M+H + =620.1

[0330] Step 7: Synthesis of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidine-3-ol carboxylate (8)

[0331] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-f, 32 mg, 0.052 mmol) was dissolved in tetrahydrofuran (2 mL), and palladium on carbon (20 mg) was added. The reaction was carried out at room temperature for 4 hours under hydrogen protection. The mixture was then filtered with diatomaceous earth, the filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol carboxylate (8, 4.09 mg, yield 12.6%). ESI[M+H] + =624.2. 1H NMR (400MHz, DMSO-d6) δ8.20 (s, 1H), 7.73 (dd, J=9.0, 6.2Hz, 1H), 7.39-7.23 (m, 2H), 7.12-6.89 (m, 1H ), 5.25 (d, J = 54.2Hz, 1H), 4.15-4.08 (m, 1H), 4.00 (t, J = 9.2Hz, 2H), 3.90 (dd, J = 9.0, 2.4Hz, 3H), 3.80 (d, J=13.2Hz, 1H), 3.72-3.57 (m, 2H), 3.08-2.99 (m, 4H), 2.80 (q, J=8.0, 7.7Hz, 1H), 2.43-2.22 (m, 2H ), 2.12-2.02(m, 2H), 2.00-1.89(m, 2H), 1.87-1.72(m, 4H), 1.58-1.41(m, 2H), 0.77(q, J=7.8Hz, 3H).

[0332] Example 9: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-ol formate (9)

[0333]

[0334] Step 1: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9-a)

[0335] Dissolve (1-(morpholinomethyl)cyclopropyl)methanol (200 mg, 1.17 mmol) in ultradry tetrahydrofuran (2 mL), add sodium hydrogen (19 mg, 0.78 mmol), stir at 0 °C for 30 minutes, then add a solution of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (8-c, 280 mg, 0.39 mmol) dissolved in tetrahydrofuran (2 mL). React at room temperature for 30 minutes. The product was quenched with water, extracted with ethyl acetate, and concentrated under reduced pressure to give the crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9-a, 200 mg, yield 18.4%). ESI[M+H + =832.1

[0336] Step 2: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9-b)

[0337] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9-a, 200 mg, 0.24 mmol) was dissolved in N,N dimethylformamide (3 mL), and cesium fluoride (218 mg, 1.44 mmol) was added. The reaction was carried out at room temperature for 1 hour. The extract was obtained by adding saturated brine and extracting with ethyl acetate. After concentration under reduced pressure, the crude product (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9-b, 214 mg, yield 26.4%) was obtained. (ESI[M+H) + =676.3

[0338] Step 3: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-ol formate (9)

[0339] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9-b, 214 mg, 0.32 mmol) was dissolved in acetonitrile (4 mL), and 1,4-dioxane hydrochloride solution (2 mL) was added. The reaction was carried out at room temperature for 1 hour. The pH was adjusted to 7 with a saturated sodium bicarbonate solution. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol carboxylate (9, 1.88 mg, yield 9.3%). ESI[M+H] + =632.3, 1 H NMR (400MHz, DMSO-d6) δ8.26 (s, 1H), 7.97 (dd, J=9.2, 6.0Hz, 1H), 7.46 (td, J=9.0, 2.5Hz, 1H), 7.38 (d, J=2.6Hz, 1H), 7.23(dd, J=18.0, 2.6Hz, 1H), 4.95(s, 1H), 4.31-4.21(m, 2H), 4.13-3.98(m, 2H), 3.88(d, J=2.0Hz, 4H), 3.74 (s, 1H), 3.64 (s, 2H), 3.21 (d, J=7.8Hz, 2H), 2.93 (dd, J=13.0, 9.0Hz, 2H), 2.38 (s, 4 H), 2.29 (d, J=2.6Hz, 2H), 1.64-1.51 (m, 2H), 1.23 (s, 2H), 0.63 (d, J=5.0Hz, 2H), 0.42 (d, J=4.6Hz, 2H).

[0340] Example 10: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10)

[0341]

[0342] Step 1: Synthesis of 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-a)

[0343] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 600 mg, 2.14 mmol) was dissolved in acetonitrile (30 ml), and then phosphorus oxychloride (986 mg, 6.43 mmol) and N,N-diisopropylethylamine (1.68 g, 13 mmol) were added. After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (840 mg, 1.605 mmol) and 3-methylpiperidine-3-ol hydrochloride (324 mg, 2.14 mmol) were added. The reaction was then carried out at 0 °C for 0.5 hours. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 30%–35%) to obtain the target product 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-a, 430 mg, yield 52.3%). ESI [M+H] + =377.4

[0344] Step 2: Synthesis of 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-b)

[0345] 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-a, 430 mg, 1.14 mmol) was dissolved in toluene (20 ml) and water (2 ml), followed by the addition of methylboronic acid (341 mg, 5.7 mol), potassium phosphate (726 mg, 3.42 mmol), and [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride (167 mg, 0.23 mmol). The mixture was heated to 105 °C and reacted for 16 h. After concentration under reduced pressure, the product was purified by TLC (petroleum ether:ethyl acetate = 2:1) to obtain the target product 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-b, 230 mg, yield 56.5%). ESI[M+H] + =357.1

[0346] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-c)

[0347] 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-b, 230 mg, 0.72 mmol), potassium carbonate (400 mg, 2.88 mmol), tetra(triphenylphosphine)palladium (166 mg, 0.144 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (553 mg, 1.08 mmol) were dissolved in 1,4-dioxane (10 ml) and water (2.5 ml), purged with nitrogen for 1 minute, and reacted in a microwave at 135 °C for 1 hour. After concentration under reduced pressure, TLC (petroleum ether: ethyl acetate = 1:1) yielded the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-c, 450 mg, yield 88.4%).

[0348] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-d)

[0349] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-c, 450 mg, 0.64 mmol) and m-chloroperoxybenzoic acid (228 mg, 0.95 mmol) were dissolved in dichloromethane (8 ml). The reaction was carried out at room temperature for 0.5 h. Quenching with water, extraction with dichloromethane, collection of the organic phase and washing with water in saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-d, 423 mg, yield 96.8%).

[0350] Step 5: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-e)

[0351] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methanol (282 mg, 1.77 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (47 mg, 1.17 mmol) was added. After reacting for another 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-d, 423 mg, 0.5 ml) was added. 9 mmol), reacted for 0.5 h. Quenched with water, extracted with ethyl acetate, the organic phase was collected and washed with water in saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-e, 298 mg, yield 22.9%). ESI[M+H + =818.1

[0352] Step 6: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-f)

[0353] (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-e, 298 mg, 0.36 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (328 mg, 2.16 mmol) was added. The mixture was reacted at 25 °C. The reaction mixture should be kept for 1 hour. Water (20 ml) was added to the reaction solution, and the mixture was extracted three times with 20 ml of ethyl acetate each time. The organic phase was collected and distilled under reduced pressure to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-f, 151 mg, yield 63.4%). ESI[M+H + =661.1

[0354] Step 7: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5HH)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10)

[0355] 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-f, 151 mg, 0.23 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (1 ml) was added at 25 °C. The reaction was allowed to proceed for 1 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10, 40 mg, yield 28.2%). ESI[M+H + =618.4, 1H NMR (400MHz, DMSO-d6) δ10.12 (d, J=25.3Hz, 1H), 7.96 (dd, J=9.1, 5.9Hz, 1H), 7.65-7.53 (m, 1H), 7.44 (t, J=9.1H z, 1H), 7.38 (d, J = 2.7Hz, 1H), 7.29 (d, J = 2.6Hz, 1H), 4.63 (d, J = 11.3Hz, 1H), 4.10 (s, 2H), 3.60 (d, J = 13.2Hz, 1H), 3.40 (d, J=13.1Hz, 1H), 3.09 (t, J=12.7Hz, 4H), 2.65 (d, J=39.6Hz, 1H), 2.56 (s, 2H), 2.05 (d, J=43.1Hz, 5H), 1.8 3 (d, J=52.0Hz, 3H), 1.68-1.60 (m, 2H), 1.51 (d, J=13.0Hz, 1H), 1.23 (s, 1H), 1.12 (d, J=17.0Hz, 1H), 1.00 (s, 3H).

[0356] Example 11: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy-)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (11)

[0357]

[0358] 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10, 22 mg, 0.036 mmol) was dissolved in methanol (1 ml), 5 mg of palladium on carbon was added, hydrogen gas was purged, and the reaction was carried out at room temperature for 2 h. The mixture was then filtered through diatomaceous earth. The filtrate was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (11, 10 mg, yield 86.5%). ESI [M+H] + =622.5, 1H NMR (400MHz, DMSO-d6) δ9.90 (d, J=19.0Hz, 1H), 7.75 (q, J=7.3Hz, 1H), 7.33 (d, J=13 .7Hz, 2H), 7.13-6.97 (m, 1H), 5.28 (d, J=53.9Hz, 1H), 4.28-4.00 (m, 4H), 3.18-3.02 (m, 4H), 2.84 (t, J=8.4Hz, 1H), 2.65-2.57 (m, 3H), 2.19-2.00 (m, 6H), 1.91-1.75 (m, 4H), 1.35 (s, 2H), 1.23 (s, 2H), 1.03 (d, J=35.8Hz, 2H), 0.75 (q, J=10.2, 7.4Hz, 4H).

[0359] Example 12: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12)

[0360]

[0361] Step 1: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-a)

[0362] (1-(morpholinomethyl)cyclopropyl)methanol (257 mg, 1.5 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (60 mg, 1.5 mmol) was added. After reacting for 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (10-d, 360 mg, 0.5 mmol) was added, and the reaction was carried out for 0.5 h. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-a, 389 mg, yield 93.7%). ESI[M+H + =830.2

[0363] Step 2: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-b)

[0364] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-a, 389 mg, 0.47 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (428 mg, 2.82 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-b, 118 mg, yield 37.2%). ESI[M+H + =672.8

[0365] Step 3: 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-c)

[0366] 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-b, 118 mg, 0.18 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (1 ml) was added at 25 °C. The reaction was allowed to proceed for 1 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-c, 21 mg, yield 19.1%). ESI [M+H] + =629.7

[0367] Step 4: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12)

[0368] 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12-c, 21 mg, 0.033 mmol) was dissolved in methanol (1 ml), 5 mg of palladium on carbon was added, and the mixture was purged with hydrogen gas. The reaction was carried out at room temperature for 2 h, and the mixture was filtered through diatomaceous earth. The filtrate was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (12, 12 mg, yield 57.1%). ESI[M+H] + =634.5, 1H NMR (600MHz, DMSO-d6) δ9.90 (s, 1H), 7.75 (d, J=7.5Hz, 1H), 7.33 (d, J=16.8H z, 2H), 7.17-6.95 (m, 1H), 4.89-4.47 (m, 1H), 4.39-4.17 (m, 3H), 3.55 (dd, J= 38.4, 15.9Hz, 6H), 2.73-2.56(m, 4H), 2.33(d, J=53.9Hz, 7H), 1.80-1.47(m, 3H), 1.03 (d, J=35.1Hz, 3H), 0.75 (q, J=7.5Hz, 3H), 0.64 (s, 2H), 0.42 (s, 2H).

[0369] Example 13: Synthesis of 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)piperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (13)

[0370]

[0371] Step 1: Synthesis of (1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-a)

[0372] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 270 mg, 0.96 mmol) was dissolved in acetonitrile (15 ml), and then phosphorus oxychloride (440 mg, 2.89 mmol) and N,N-diisopropylethylamine (746 mg, 5.79 mmol) were added. After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (373 mg, 2.89 mmol) and 3-methylpiperidine-3-ol hydrochloride (110 mg, 0.96 mmol) were added. The reaction was then carried out at 0 °C for 0.5 hours. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 30%–35%) to obtain the target product (1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-a, 190 mg, yield 52.0%). ESI [M+H]+ = 377.5

[0373] Step 2: Synthesis of (1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-b)

[0374] (1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-a, 190 mg, 0.5 mmol), toluene (10 mL), and water (1 mL) were added, followed by the addition of methylboric acid (120 mg, 2.0 mol), potassium phosphate (318 mg, 1.5 mmol), and [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride (10 mg, 0.01 mmol). The mixture was heated to 105 °C and reacted for 16 h. After concentration under reduced pressure, the mixture was purified by TLC (petroleum ether: ethyl acetate = 2:1) to obtain the target product (1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-b, 96 mg, yield 53.9%). ESI [M+H] + =357.7

[0375] Step 3: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-c)

[0376] (1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-b, 96 mg, 0.27 mmol), potassium carbonate (149 mg, 1.08 mmol), tetraphenylphosphine palladium (62 mg, 0.054 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (278 mg, 0.54 mmol) were dissolved in 1,4-dioxane (5 ml) and water (1.25 ml), purged with nitrogen for 1 minute, and reacted in a microwave at 135 °C for 1 hour. After concentration under reduced pressure, TLC (petroleum ether: ethyl acetate = 1:1) yielded the target product (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-c, 107 mg, yield 56.3%).

[0377] Step 4: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-d)

[0378] (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-c, 107 mg, 0.15 mmol) and m-chloroperoxybenzoic acid (39 mg, 0.23 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 0.5 h. Quenching with water, extraction with dichloromethane, collection of the organic phase and washing with water in saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-d, 97 mg, yield 53.9%).

[0379] Step 5: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-e)

[0380] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methanol (64 mg, 0.402 mmol) was dissolved in anhydrous tetrahydrofuran (2 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (47 mg, 0.27 mmol) was added. After reacting for 0.5 hours, (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-b, 97 mg, 0.134 mmol) was added, and the reaction was carried out for 0.5 h. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered and concentrated under reduced pressure to obtain the target product (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-e, 65 mg, yield 59.1%). ESI[M+H + =818.2

[0381] Step 6: Synthesis of (1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (13-f)

[0382] (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl)methanol (13-e, 65 mg, 0.08 mmol) was dissolved in N,N-dimethylformamide (2 ml), and cesium fluoride (73 mg, 0.48 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to obtain the target product (1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (13-f, 25 mg, yield 47.2%). ESI[M+H +=662.1

[0383] Step 7: Synthesis of 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)piperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (13)

[0384] (1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)3-methylpiperidin-3-ol (13-f, 25 mg, 0.04 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution was added at 25 °C. (0.5 ml), react for 1 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)piperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (13.5 mg, yield 20.2%). ESI[M+H + =618.4

[0385] Example 14: Synthesis of (3R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxy-7,8-dihydronaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14)

[0386]

[0387] Step 1: Synthesis of (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-a)

[0388] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1, 470 mg, 1.7 mmol) was dissolved in acetonitrile (20 ml), and then phosphorus oxychloride (270 mg, 5.1 mmol) and N,N-diisopropylethylamine (1315 mg, 10.2 mmol) were added. After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (658 mg, 5.1 mmol) and 3-methylpiperidine-3-ol hydrochloride (316 mg, 2.6 mmol) were added. The reaction was then carried out at 0 °C for 0.5 hours. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 30%–35%) to obtain the target product (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-a, 206 mg, yield 34.19%). ESI [M+H] + =359.1

[0389] Step 2: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-b)

[0390] (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-a, 200 mg, 0.57 mmol), potassium carbonate (314 mg, 2.28 mmol), tetrakis(triphenylphosphine)palladium (70 mg, 0.06 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (402 mg, 0.86 mmol) were dissolved in 1,4-dioxane (10 ml) and water (2.5 ml), purged with nitrogen for 1 minute, and reacted in a microwave oven at 135 °C for 1 hour. After concentration under reduced pressure, TLC (petroleum ether: ethyl acetate = 3:1) yielded the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-b, 330 mg, yield 81.7%).

[0391] Step 3: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-c)

[0392] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-b, 330 mg, 0.47 mmol) and m-chloroperoxybenzoic acid (122 mg, 0.71 mmol) were dissolved in dichloromethane (8 ml). The reaction was carried out at room temperature for 0.5 h. Quenching with water, extraction with dichloromethane, collection of the organic phase and washing with water in saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidine-3-ol (14-c, 300 mg, yield 87.5%).

[0393] Step 4: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3d]pyrimidin-4-yl)piperidin-3-ol (14-d)

[0394] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolin-7a(5H)-yl)methanol (196 mg, 1.23 mmol) was dissolved in anhydrous tetrahydrofuran (2 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (20 mg, 0.82 mmol) was added. After reacting for 0.5 hours, (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-c, 300 mg, 0.41 mmol) was added, and the reaction was carried out for 0.5 h. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered and concentrated under reduced pressure to give the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-d, 80 mg, yield 24.1%). ESI[M+H + =820.6

[0395] Step 5: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-e)

[0396] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-d, 80 mg, 0.098 mmol) was dissolved in N,N-dimethylformamide (3 ml), and cesium fluoride (89 mg, 0.585 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to give the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-e, 46 mg, yield 69.7%). ESI[M+H+ =664.2

[0397] Step 6: (3R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxy-7,8-dihydronaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14)

[0398] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14-e, 46 mg, 0.07 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (1 ml) was added at 25 °C. The reaction was allowed to proceed for 1 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product (3R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxy-7,8-dihydronaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (14,11 mg, yield 25.8%). ESI [M+H]+ = 620.2. 1 H NMR (400MHz, DMSO-d6) δ8.16 (s, 1H), 7.96 (dd, J=9.2, 6.0Hz, 1H), 7.45 (td, J=9.0, 2.6Hz, 1H), 7.36 (dd , J=11.4, 2.5Hz, 1H), 7.23 (dd, J=18.9, 2.6Hz, 1H), 5.35 (d, J=3.8Hz, 1H), 5.23-5.19 (m, 1H), 4.18-4.07 (m, 2H), 4.05-3.96 (m, 3H), 3.88 (d, J=1.6Hz, 5H), 3.20 (dd, J=12.9, 7.0Hz, 2H), 3.14-3.07 (m, 2H), 3.02 (d, J=2.7Hz, 1H), 2.84 (s, 1H), 2.13 (s, 1H), 2.06 (d, J=5.5Hz, 2H), 1.76 (s, 5H), 1.48 (d, J=36.7Hz, 2H).

[0399] Example 15: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15)

[0400]

[0401] Step 1: Preparation of 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-a)

[0402] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 400 mg, 1.45 mmol) was dissolved in acetonitrile (10 ml), and then phosphorus oxychloride (1.67 g, 10.9 mmol) and N,N-diisopropylethylamine (1.69 g, 13.05 mmol) were added. After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (2.81 g, 21.75 mmol) and 3-methylpiperidine-3-ol hydrochloride (350 mg, 2.31 mmol) were added. The reaction was carried out at 0 °C for 0.5 hours. Preparation of the target product 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-a, 383 mg, yield 70.93%) after flash purification (petroleum ether:ethyl acetate = 4:1) following vacuum concentration. (ESI)[M+H + =375.1, 377.0

[0403] Step 2: Preparation of 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-b)

[0404] 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-a, 200 mg, 0.53 mmol) was dissolved in toluene (8 ml) and water (0.8 ml), and then methylboronic acid (159 mg, 2.65 mmol), potassium phosphate (450 mg, 2.12 mmol), and [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride (78 mg, 0.106 mmol) were added. The mixture was heated to 105 °C and reacted for 24 h. After concentration under reduced pressure, TLC (dichloromethane:methanol = 100:3) purification yielded the target product 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-b, 80 mg, yield 42.33%). (ESI)[M+H + =357.1, 359.1

[0405] Step 3: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-c)

[0406] 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-b, 80 mg, 0.224 mmol), potassium carbonate (124 mg, 0.896 mmol), tetrakis(triphenylphosphine)palladium (52 ​​mg, 0.0448 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (138 mg, 0.269 mmol) were dissolved in 1,4-dioxane (3 ml) and water (0.75 ml), purged with nitrogen for 1 minute, and microwaved at 135 °C for 1.5 hours. Preparation of the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol by TLC (15-c, 90 mg, yield 56.96%) after concentration under reduced pressure.

[0407] Step 4: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-d)

[0408] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-c, 90 mg, 0.127 mmol) and m-chloroperoxybenzoic acid (66 mg, 0.381 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 1 h. Quenching with aqueous solution, extraction with dichloromethane, collection of organic phase and washing with saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-d, 92 mg, yield 97.87%).

[0409] Step 5: Preparation of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-e)

[0410] (1-(morpholinomethyl)cyclopropyl)methanol (63 mg, 0.366 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (15 mg, 0.366 mmol) was added. After reacting for another 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-d) was added. 90 mg, 0.122 mmol), reacted for 0.5 h, concentrated under reduced pressure, and purified by TLC (dichloromethane:methanol = 10:1) to obtain the target product (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-e, 32 mg, yield 31.68%). (ESI)[M+H) + =830.9, 831.9.

[0411] Step 6: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-f)

[0412] (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-e, 32 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (2 ml), and cesium fluoride (36 mg, 0.24 mmol) was added. The solution was incubated at 25°C. The reaction was carried out for 0.5 hours. Water (20 ml) was added to the reaction solution, and the mixture was extracted three times with 20 ml of ethyl acetate each time. The organic phase was collected and distilled under reduced pressure to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-f, 30 mg). (ESI)[M+H] + =674.6, 665.7

[0413] Step 7: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15-f, 25 mg, 0.04 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (0.5 ml) was added at 25 °C. The reaction was carried out for 0.5 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (15, 5.27 mg, yield 21.08%). (ESI)[M+H + =630.3, 631.3 1H NMR (400MHz, DMSO-d6) δ10.15 (d, J=16.0Hz, 1H), 7.97 (dd, J=9.2, 5.9Hz, 1H), 7.45 (t, J=9.0Hz, 1H), 7.38 (d, J=2.6Hz, 1H), 7.29 (dd, J=7.1, 2.6Hz, 1H), 4.62 (d, J=12.2Hz, 1H), 4.30 (td, J=31.6, 10.2Hz, 3H), 4.1 2(s, 1H), 3.60(d, J=46.8Hz, 6H), 3.41(s, 1H), 3.08(s, 1H), 2.69-2.61(m, 1H), 2.56(s, 2H), 2.44-2.24(m , 6H), 1.64 (s, 1H), 1.49 (d, J=13.2Hz, 1H), 1.23 (s, 3H), 1.00 (s, 2H), 0.64 (d, J=11.2Hz, 2H), 0.41 (s, 2H).

[0414] Example 16: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)aza-4-ol (16)

[0415]

[0416] Step 1: Preparation of 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-a)

[0417] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1, 470 mg, 1.705 mmol) was dissolved in acetonitrile (5 ml), and then phosphorus oxychloride (805 mg, 5.25 mmol) and N,N-diisopropylethylamine (881 mg, 6.82 mmol) were added. The reaction was carried out at 80 °C for 0.5 hours, and then the reaction was cooled to 0 °C. Then N,N-diisopropylethylamine (881 mg, 6.82 mmol) and Azerbaijan-4-ol hydrochloride (350 mg, 2.308 mmol) were added, and the reaction was carried out at 0 °C for 0.5 hours. After concentration under reduced pressure, TLC (methanol:dichloromethane = 2:25) purification yielded the target product 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-a, 448 mg, yield 70.44%). (ESI)[M+H +=373.4, 375.4

[0418] Step 2: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-b)

[0419] 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-a, 440 mg, 1.18 mmol), potassium carbonate (652 mg, 4.72 mmol), tetrakis(triphenylphosphine)palladium (273 mg, 0.236 mmol), and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (726 mg, 1.416 mmol) were dissolved in 1,4-dioxane (10 ml) and water (2.5 ml), purged with nitrogen for 1 minute, and microwaved at 135 °C for 1.5 hours. After concentration under reduced pressure, TLC (dichloromethane:methanol = 15:1) yielded the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-b, 602 mg, yield 70.57%).

[0420] Step 3: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-c)

[0421] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-b, 150 mg, 0.207 mmol) and m-chloroperoxybenzoic acid (107 mg, 0.621 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 1 h. Quenching with water, extraction with dichloromethane, collection of the organic phase and washing with water in saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-c, 100 mg, yield 64.10%).

[0422] Step 4: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-d)

[0423] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (51 mg, 0.318 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (13 mg, 0.318 mmol) was added. After reacting for another 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)zepam-4- Alcohol (16-c, 80 mg, 1.06 mmol) was reacted for 0.5 h, concentrated under reduced pressure, and purified by TLC (dichloromethane:methanol = 10:1) to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-d, 65 mg, yield 73.86%). (ESI)[M+H + =834.3, 835.4

[0424] Step 5: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)aza-4-ol (16)

[0425] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-d, 35 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (2 ml), and cesium fluoride (36 mg, 0.24 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected, and the crude product was purified by preparative liquid chromatography after vacuum distillation to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridano[4,3-d]pyrimidin-4-yl)aza-4-ol (16, 4.45 mg, yield 16.48%). (ESI)[M+H + =678.3, 679.3 1 H NMR (400MHz, DMSO-d6) δ8.16-8.09 (m, 1H), 7.76 (d, J=2.6Hz, 1H), 7.62-7.55 (m, 1H) , 7.46 (d, J = 7.9Hz, 1H), 5.45-5.24 (m, 3H), 4.71-4.60 (m, 1H), 4.09 (d, J = 15.0Hz, 1H) , 3.90 (s, 3H), 3.72 (s, 1H), 3.61 (s, 1H), 3.48 (d, J = 1.9Hz, 2H), 2.12-1.98 (m, 4H), 1 .72 (s, 4H), 1.51 (s, 2H), 1.37 (s, 1H), 1.26 (d, J = 3.7Hz, 10H), 0.89 (t, J = 6.6Hz, 1H).

[0426] Example 17: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17)

[0427]

[0428] Step 1: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-a)

[0429] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (16-b, 237 mg, 0.328 mmol) and m-chloroperoxybenzoic acid (170 mg, 0.984 mmol) were dissolved in dichloromethane (10 ml). The reaction was carried out at room temperature for 1 h. Quenching with water, extraction with dichloromethane, collection of the organic phase and washing with water in saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-a, 280 mg).

[0430] Step 2: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-b)

[0431] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (63 mg, 0.396 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (16 mg, 0.396 mmol) was added. After reacting for another 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol was added. (17-a, 100 mg, 0.132 mmol), after reacting for 0.5 h, the product was concentrated under reduced pressure and purified by TLC (dichloromethane:methanol = 10:1) to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-b, 48 mg, yield 43.64%). (ESI)[M+H + =834.3, 835.4

[0432] Step 3: Preparation of 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-c)

[0433] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-b, 48 mg, 0.058 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (0.5 ml) was added at 25 °C. The reaction was allowed to proceed for 0.5 h. The reaction solution was concentrated under reduced pressure to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridano[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-c, 52 mg, yield 113%). (ESI)[M+H + =790.4, 791.3

[0434] Step 4: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17)

[0435] 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17-c, 52 mg, 0.066 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (60 mg, 0.396 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected, and the crude product was purified by preparative liquid chromatography after vacuum distillation to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (17, 6.97 mg, yield 16.59%). (ESI)[M+H + =634.2, 635.2 1 HNMR (400MHz, DMSO-d6) δ10.17 (d, J=3.8Hz, 1H), 7.97 (dd, J=9.2, 5.9Hz, 1H), 7.46 (td, J=9.1, 1.7H z, 1H), 7.38 (d, J=2.6Hz, 1H), 7.23 (dd, J=8.7, 2.6Hz, 1H), 5.36 (s, 1H), 5.22 (s, 1H), 4.62 (dd, J=12 .4, 3.6Hz, 1H), 4.10 (s, 1H), 4.01 (d, J = 16.6Hz, 2H), 3.86 (s, 5H), 3.71 (d, J = 19.7Hz, 2H), 3.57 (s, 2 H), 3.11 (s, 2H), 3.03 (s, 1H), 2.84 (s, 1H), 2.13 (s, 1H), 2.03 (d, J=20.1Hz, 4H), 1.87-1.65 (m, 6H).

[0436] Example 18: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18)

[0437]

[0438] Step 1: Preparation of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18-a)

[0439] (1-(morpholinomethyl)cyclopropyl)methanol (122 mg, 0.714 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C, and 60% sodium hydride (29 mg, 0.714 mmol) was added. After reacting for another 0.5 hours, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d] The target product, 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18-a, 272 mg), was reacted with pyrimidin-4-yl)aza-4-ol (17-a, 180 mg, 0.238 mmol) for 0.5 h. The mixture was then concentrated under reduced pressure to obtain the crude product. (ESI)[M+H] + =846.4, 847.4

[0440] Step 2: Preparation of 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18-b)

[0441] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18-a, 272 mg, 0.32 mmol) was dissolved in acetonitrile (10 ml), and 1,4-dioxane hydrochloride solution (1 ml) was added at 25 °C. The reaction was allowed to proceed for 0.5 h. The reaction solution was concentrated under reduced pressure to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18-b, 115 mg, yield 44.75%). (ESI)[M+H + =802.9, 803.9

[0442] Step 3: Preparation of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18)

[0443] 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18-b, 115 mg, 0.14 mmol) was dissolved in N,N-dimethylformamide (3 ml), and cesium fluoride (213 mg, 1.4 mmol) was added. The mixture was reacted at 25 °C for 1 hour. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected, and the crude product was purified by preparative liquid chromatography after vacuum distillation to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)zepam-4-ol (18, 1.23 mg, yield 1.37%). (ESI)[M+H + =646.3, 647.3 1H NMR (400MHz, DMSO-d6) δ10.17 (d, J=3.8Hz, 1H), 7.97 (dd, J=9.2, 5.9Hz, 1H), 7.46 (td, J=9.1, 1.7H z, 1H), 7.38 (d, J=2.6Hz, 1H), 7.23 (dd, J=8.7, 2.6Hz, 1H), 5.36 (s, 1H), 5.22 (s, 1H), 4.62 (dd, J=12 .4, 3.6Hz, 1H), 4.10 (s, 1H), 4.01 (d, J = 16.6Hz, 2H), 3.86 (s, 5H), 3.71 (d, J = 19.7Hz, 2H), 3.57 (s, 2 H), 3.11 (s, 2H), 3.03 (s, 1H), 2.84 (s, 1H), 2.13 (s, 1H), 2.03 (d, J=20.1Hz, 4H), 1.87-1.65 (m, 6H).

[0444] Example 19: Preparation of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19)

[0445]

[0446] Step 1: Preparation of (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-a)

[0447] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 1 g, 3.57 mmol) was dissolved in acetonitrile (20 ml), and then phosphorus oxychloride (1.64 g, 10.71 mmol) and N,N-diisopropylethylamine (2.77 g, 21.42 mmol) were added. After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (2.77 g, 21.42 mmol) and (R)-3-methylpiperidin-3-ol hydrochloride (800 mg, 5.28 mmol) were added. The reaction was carried out at 0 °C for 0.5 hours. After concentration under reduced pressure, flash purification (petroleum ether:ethyl acetate = 4:1) yielded the target product (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-a, 730 mg, yield 54.28%). (ESI)[M+H +=377.2, 379.0

[0448] Step 2: Preparation of (R)-1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-b)

[0449] (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-a, 480 mg, 1.27 mmol) was dissolved in toluene (6 ml) and water (0.6 ml), and then methylboronic acid (304 mg, 5.08 mmol), potassium phosphate (1.08 g, 5.08 mmol), and 1,1′-bis(diphenylphosphino)ferrocene palladium dichloride (II) (186 mg, 0.254 mmol) were added. The mixture was heated to 105 °C and reacted for 24 h. After concentration under reduced pressure, TLC (dichloromethane:methanol = 100:4) purification yielded the target product (R)-1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-b, 100 mg, yield 22.08%). (ESI)[M+H + =357.2

[0450] Step 3: Preparation of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-c)

[0451] (R)-1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-b, 100 mg, 0.28 mmol), potassium carbonate (155 mg, 1.12 mmol), tetrakis(triphenylphosphine)palladium (65 mg, 0.056 mmol), and ((2-fluoro-6-(methoxymethyloxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (172 mg, 0.336 mmol) were dissolved in 1,4-dioxane (4 ml) and water (1 ml), purged with nitrogen for 1 minute, and microwaved at 135 °C for 1.5 hours. After concentration under reduced pressure, TLC (dichloromethane:methanol = 15:1) yielded the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-c, 123 mg, yield 62.12%).

[0452] Step 4: Preparation of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-d)

[0453] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-c, 120 mg, 0.17 mmol) and m-chloroperoxybenzoic acid (88 mg, 0.51 mmol) were dissolved in dichloromethane (5 ml). The reaction was carried out at room temperature for 1 h. Quenching with aqueous solution, extraction with dichloromethane, collection of organic phase and washing with saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-d, 127 mg, 100% yield).

[0454] Step 5: Preparation of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-e)

[0455] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (77 mg, 0.486 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 pyr, and 60% sodium hydride (20 mg, 0.486 mmol) was added. After reacting for another 0.5 hours, (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol was added. (19-d, 120 mg, 0.162 mmol), after reacting for 0.5 h, the product was concentrated under reduced pressure and purified by TLC (dichloromethane:methanol = 10:1) to obtain the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-e, 57 mg, yield 42.86%). (ESI)[M+H + =818.6, 819.6

[0456] Step 6: Preparation of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-f)

[0457] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-e, 32 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (64 mg, 0.42 mmol) was added. The mixture was reacted at 25 °C for 0.5 h. The reaction mixture was added to water (20 ml), and extracted three times with ethyl acetate (20 ml each time). The organic phase was collected and distilled under reduced pressure to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-f, 60 mg). (ESI)[M+H] + =662.6, 663.6

[0458] Step 7: Preparation of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19)

[0459] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19-f, 60 mg, 0.09 mmol) was dissolved in acetonitrile (5 ml), and 1,4-dioxane hydrochloride solution (0.5 ml) was added at 25 °C. The reaction was allowed to proceed for 0.5 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (19,60 mg, yield 15.21%). (ESI)[M+H + =618.5, 619.4 1H NMR (400MHz, DMSO-d6) δ10.17 (s, 1H), 7.97 (dd, J=9.2, 5.9Hz, 1H), 7.45 (t, J=9.0Hz, 1H), 7.39 (d, J=2.6Hz, 1H), 7.29 (d, J =2.5Hz, 1H), 5.35 (s, 1H), 5.22 (s, 1H), 4.65-4.58 (m, 2H), 4.13 (d, J = 11.7Hz, 2H), 3.98 (d, J = 10.4Hz, 1H), 3.60 (d, J = 13.1 Hz, 1H), 3.41 (s, 1H), 3.13-3.07 (m, 3H), 3.02 (s, 1H), 2.86-2.80 (m, 1H), 2.69-2.61 (m, 1H), 2.56 (s, 3H), 2.13 (s, 1H), 2.0 7-2.05 (m, 1H), 2.00 (s, 1H), 1.86-1.81 (m, 1H), 1.80-1.75 (m, 2H), 1.67-1.62 (m, 2H), 1.50 (d, J=13.0Hz, 1H), 1.00 (s, 3H).

[0460] Example 20: Synthesis of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0461]

[0462] Step 1: Synthesis of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (20)

[0463] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (9.32 mg, 0.05 mmol) was dissolved in tetrahydrofuran (5 mL), and palladium on carbon (30 mg) was added. The reaction was carried out at room temperature for 2 hours under hydrogen protection. Then, diatomaceous earth was added and filtered. The filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (20.89 mg, yield 15.4%). ESI[M+H] + =636.3 1 H NMR(400MHz, DMSO-d6)δ 9.81-9.89(m, 1H), 7.75-7.71(m, 1H), 7.37-7.22(m, 2H), 7.1-6.88(m, 1H), 4.92( s, 1H), 4.36-4.25 (m, 2H), 4.00 (s, 1H), 3.91-3.89 (m, 3H), 3.80 (s, 1H), 3.66-3.65 (m, 1H), 3.49 (s, 4H), 3.03-2.99 (m, 1H), 2.35 (s, 4H), 2.27 (s, 2H), 1.91-1.84 (m, 2H), 1.53-1.45(m, 2H), 1.21(s, 1H), 0.83-0.76(m, 3H), 0.62(s, 2H), 0.39(s, 2H).

[0464] Example 21: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate

[0465]

[0466] Step 1: Synthesis of (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-a)

[0467] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a, 606.5 mg, 2.2 mmol) was dissolved in acetonitrile (16 mL), followed by the addition of N,N-diisopropylethylamine (2.3 mL, 13.2 mmol) and phosphorus oxychloride (0.62 mL, 6.6 mmol). After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (2.3 mL, 13.2 mmol) and (R)-3-methylpiperidin-3-ol (400 mg, 2.64 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–40%) to obtain the target product (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-a, 293 mg, yield 25.0%). ESI[M+H + =373.0

[0468] Step 2: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-b)

[0469] (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-a, 293 mg, 0.78 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphthyl)ethynyl)triisopropylsilane (600 mg, 1.17 mmol), and potassium carbonate (431 mg, 3.12 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (12 mL), and then tetra(triphenylphosphine)palladium (90 mg, 0.078 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-b, 420 mg) was obtained by thin-layer chromatography with silica gel plates (petroleum ether: ethyl acetate = 3:1).

[0470] Step 3: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-c)

[0471] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-b, 420 mg, 0.58 mmol) was dissolved in dichloromethane (5 mL), and then m-chloroperoxybenzoic acid (150 mg, 0.87 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to obtain crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-c, 470 mg).

[0472] Step 4: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0473] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (296 mg, 1.86 mmol) was dissolved in ultra-dry tetrahydrofuran (3 mL), sodium hydrogen (30 mg, 1.24 mmol) was added, and the mixture was stirred at 0 °C for 30 minutes. Then, a solution of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21-c, 470 mg, 0.62 mmol) dissolved in tetrahydrofuran (3 mL) was added. The reaction was carried out at room temperature for 30 minutes. Quenching with water, extraction with ethyl acetate, and concentration under reduced pressure yielded the crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(21-d, 798 mg). ESI[M+H] + =834.2

[0474] Step 5: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (21-e)

[0475] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (21-d, 798 mg, 0.96 mmol) was dissolved in N,N-dimethylformamide (4 mL), and cesium fluoride (875 mg, 5.76 mmol). The reaction was carried out at room temperature for 1 hour. Saturated brine was added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, the crude product (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(21-e, 810 mg). ESI[M+H] + =678.2

[0476] Step 6: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate (21)

[0477] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphthyl-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(21-e, 810 mg, 1.2 mmol) was dissolved in acetonitrile (5 mL), and 1,4-dioxane hydrochloride solution (1.5 mL) was added. The reaction was carried out at room temperature for 1 hour. The pH was adjusted to saturated sodium bicarbonate solution. 7. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21,70 mg, yield 9.2%). ESI[M+H + =634.2 1H NMR(400MHz, DMSO-d6)δ 8.12(s, 1H), 7.95-7.90(m, 1H), 7.45-7.39(m, 1H), 7.38-7.32(m, 1H), 7.25-7.16( m, 1H), 5.33 (s, 0.5H), 5.19 (s, 0.5H), 4.56-4.36 (m, 2H), 4.20-4.09 (m, 2H), 4.02- 3.97(m, 1H), 3.66-3.63(m, 3H), 3.70-3.62(m, 2H), 3.15-3.02(m, 4H), 2.85-2.75( m, 1H), 2.15-1.95 (m, 4H), 1.88-1.83 (m, 3H), 1.65-1.49 (m, 3H), 1.08-1.01 (m, 3H).

[0478] Example 22: Synthesis of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol carboxylate

[0479]

[0480] Step 1: Synthesis of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol

[0481] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (21,40 mg, 0.06 mmol) was dissolved in tetrahydrofuran (2 mL), and palladium on carbon (20 mg) was added. The reaction was carried out at room temperature for 2 hours under hydrogen protection. The mixture was then filtered with diatomaceous earth, the filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (22, 10.38 mg, yield 27.13%). ESI[M+H+ =638.3 1 H NMR(400MHz, DMSO-d6)δ 9.92-9.85(m, 1H), 7.75-7.69(m, 1H), 7.34-7.29(m, 2H), 7.13-6.95(m, 1H), 5.42(s, 0.5H), 5.29 (s, 0.5H), 4.52-4.51 (m, 1H), 4.37-4.2 (m, 2H), 4.02-4.00 (m, 1H), 3.90-3.89 (m, 3H), 3.79-3.70 (m, 1H), 3.56-3.49 (m, 2H), 3.42-3.36 (m, 2H), 3.29-3.18 (m, 2H), 2.97 (s, 1H), 2.41-2.18 (m, 3H) , 2.15-2.07(m, 1H), 2.01-1.79(m, 4H), 1.61-1.53(m, 3H), 1.32-1.20(m, 2H), 1.05-0.99(m, 3H).

[0482] Example 23: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol

[0483]

[0484] Step 1: Synthesis of 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-a)

[0485] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a910 mg, 3.3 mmol) was dissolved in acetonitrile (18 mL), followed by the addition of N,N-diisopropylethylamine (2.6 mL, 19.8 mmol) and phosphorus oxychloride (0.92 mL, 9.9 mmol). After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (2.6 mL, 19.8 mmol) and 3-methylazacyclobutane-3-ol (815.63 mg, 6.6 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–40%) to obtain the target product 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-a, 388 mg, yield 39.9%). ESI[M+H] + =345.0

[0486] Step 2: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-b)

[0487] (1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-a, 207 mg, 0.6 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphthyl)ethynyl)triisopropylsilane (461 mg, 0.9 mmol), and potassium carbonate (332 mg, 2.4 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water. Mixed solvent (6 mL), then tetrakis(triphenylphosphine)palladium (70 mg, 0.06 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (petroleum ether: ethyl acetate = 3:1) to obtain the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-b, 236 mg).

[0488] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-c)

[0489] (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-b, 236 mg, 0.34 mmol) was dissolved in dichloromethane (4 mL), and then m-chloroperoxybenzoic acid (88 mg, 0.51 mL) was added. mol). React at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to give crude product (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-c, 321 mg).

[0490] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0491] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (210 mg, 1.32 mmol) was dissolved in ultra-dry tetrahydrofuran (3 mL), sodium hydrogen (21 mg, 0.88 mmol) was added, and the mixture was stirred at 0 °C for 30 minutes. Then, (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylazine Heterocyclic butane-3-ol (23-c, 321 mg, 0.44 mmol) dissolved in tetrahydrofuran (3 mL) was added. The reaction was carried out at room temperature for 30 minutes. The reaction was quenched with water, extracted with ethyl acetate, and concentrated under reduced pressure to give the crude product (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (23-d, 330 mg). ESI[M+H] + =806.4

[0492] Step 5: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy]-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-e)

[0493] (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (23-d, 330 mg, 0.4 mmol)) was dissolved in N,N-dimethylformamide (5 mL), and cesium fluoride (365 mg, 2.4 mmol) was added. The reaction was carried out at room temperature for 1 hour. Extracted with saturated brine and ethyl acetate, the crude product was concentrated under reduced pressure to obtain 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy]-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-e, 732 mg). ESI[M+H] + =650.3

[0494] Step 6: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23)

[0495] 1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy]-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23-e, 732 mg, 1.12 mmol) was dissolved in acetonitrile (3 mL), and 1,4-dioxane hydrochloride solution (1.5 mL) was added. The reaction was carried out at room temperature for 1 minute. The reaction mixture was adjusted to pH 7 with a saturated sodium bicarbonate solution. The solution was then purified by preparative liquid chromatography to yield the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23, 45.07 mg, yield 6.6%). ESI [M+H] + =606.2 1 H NMR(400MHz, DMSO-d6)δ 10.11(s, 1H), 7.95-7.91(m, 1H), 7.45-7.40(m, 1H), 7.35-7.32(m, 1H), 7.17( s, 1H), 5.65-5.61 (m, 1H), 5.32 (s, 0.5H), 5.18 (s, 0.5H), 4.38-4.33 (m, 1H), 4. 26-4.13(m, 2H), 4.10-3.93(m, 4H), 3.85(s, 3H), 3.11-2.97(m, 3H), 2.86-2.74 (m, 1H), 2.10-2.08 (m, 1H), 2.05-1.95 (m, 2H), 1.85-1.70 (m, 3H), 1.39 (s, 3H).

[0496] Example 24: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylaminobutane-3-ol trifluoroacetate

[0497]

[0498] Step 1: Synthesis of (1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol trifluoroacetate (24)

[0499] 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (23,40 mg, 0.066 mmol) was dissolved in tetrahydrofuran (4 mL), and palladium on carbon (189 mg) was added. The reaction was carried out at room temperature for 1 hour under hydrogen protection. The mixture was then filtered with diatomaceous earth, the filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridano[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (24, 2.57 mg, yield 6.4%). ESI[M+H + =610.3 1 H NMR(400MHz, DMSO-d6)δ 10.82(s, 1H), 7.75-7.71(m, 1H), 7.34-7.24(m, 2H), 7.01-7.98(m, 1H), 5.61(s, 0.5H), 5. 59(s, 0.5H), 4.60-4.58(m, 2H), 4.44-4.38(m, 2H), 4.25-4.19(m, 1H), 4.11-4.09(m, 1H), 3.89(s, 3H), 3.83-3.66(m, 4H), 3.29-3.18(m, 2H), 2.67-2.58(m, 1H), 2.42-2.39(m, 1H), 2.28-2.26(m, 2H), 2.14-2.11(m, 2H), 2.09-1.98(m, 1H), 1.40(s, 3H), 0.87-0.77(m, 3H).

[0500] Example 25: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol trifluoroacetate

[0501]

[0502] Step 1: Synthesis of 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-a)

[0503] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a 1.16 g, 4.2 mmol) was dissolved in acetonitrile (20 mL), followed by the addition of N,N-diisopropylethylamine (4.4 mL, 25.2 mmol) and phosphorus oxychloride (1.2 mL, 12.6 mmol). After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (4.4 mL, 25.2 mmol) and aziridine-3-ol (690 mg, 6.3 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–50%) to obtain the target product 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-a, 153 mg, yield 11.0%). ESI [M+H] + =331.0

[0504] Step 2: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-b)

[0505] 1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-a, 153 mg, 0.46 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (354 mg, 0.69 mmol), and potassium carbonate (254 mg, 1.84 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (6 mL), and then tetra(triphenylphosphine)palladium (53 mg, 0.046 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-b, 295 mg) was obtained by thin-layer chromatography with silica gel plates (petroleum ether: ethyl acetate = 3:1).

[0506] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-c)

[0507] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-b, 295 mg, 0.43 mmol) was dissolved in dichloromethane (4 mL), and then m-chloroperoxybenzoic acid (111 mg, 0.645 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to obtain crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-c, 285 mg).

[0508] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0509] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (191 mg, 1.2 mmol) was dissolved in ultra-dry tetrahydrofuran (2 mL), sodium hydrogen (19.2 mg, 0.8 mmol) was added, and the mixture was stirred at 0 °C for 30 minutes. Then, a solution of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-c, 285 mg, 0.4 mmol) dissolved in tetrahydrofuran (2 mL) was added. The reaction was carried out at room temperature for 30 minutes. The sample was quenched with water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (25-d, 51 mg, yield 16.1%). ESI[M+H + =792.4

[0510] Step 5: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-e)

[0511] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (25-d, 51 mg, 0.06 mmol) was dissolved in acetonitrile (2 mL), and 1,4-dioxane hydrochloride solution (0.2 mL) was added. The reaction was carried out at room temperature for 1 minute. Hours. After concentration under reduced pressure, crude product 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-e, 59 mg). ESI [M+H] + =748.3

[0512] Step 6: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol trifluoroacetate (25)

[0513] 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25-e, 59 mg, 0.08 mmol) was dissolved in N,N dimethylformamide (2 mL), and cesium fluoride (729 mg, 60.0 mmol) was added. The reaction was carried out at room temperature for 4 hours. The sample was extracted with ethyl acetate after adding saturated brine and water, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25 mg, 1.19 mg, yield 2.51%). ESI[M+H + =592.2 1 H NMR (400MHz, DMSO-d6) δ10.78 (s, 1H), 10.14 (s, 1H), 7.96-7.92 (m, 1H), 7.45-7. 34(m, 2H), 7.19-7.16(m, 1H), 5.79-5.45(m, 2H), 4.70(s, 1H), 4.55-4.49(m, 4H) , 4.35-4.22(m, 1H), 4.04-3.95(m, 2H), 3.89(s, 3H), 3.83-3.71(m, 3H), 2.60-2. 58(m, 1H), 2.46-2.28(m, 2H), 2.25-2.19(m, 2H), 2.16-2.09(m, 1H), 1.21(s, 1H).

[0514] Example 26: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol carboxylate

[0515]

[0516] Step 1: Synthesis of (1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol carboxylate (26)

[0517] 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (25, 39 mg, 0.066 mmol) was dissolved in tetrahydrofuran (1 mL), and palladium on carbon (28 mg) was added. The reaction was carried out at room temperature for 1 hour under hydrogen protection. The mixture was then filtered with diatomaceous earth, the filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product (1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (26, 0.67 mg, yield 1.76%). ESI[M+H + =696.2

[0518] Example 27: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate

[0519]

[0520] Step 1: Synthesis of (1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-a)

[0521] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a 303 mg, 1.1 mmol) was dissolved in acetonitrile (12 mL), followed by the addition of N,N-diisopropylethylamine (1.1 mL, 6.6 mmol) and phosphorus oxychloride (0.3 mL, 3.3 mmol). After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (1.1 mL, 6.6 mmol) and aziridine-3-ylmethanol (163 mg, 1.32 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel (petroleum ether:ethyl acetate = 1:1) to obtain the target product (1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-a, 177 mg, yield 21.5%). ESI[M+H + =345.0

[0522] Step 2: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-b)

[0523] (1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-a, 150 mg, 0.435 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (334 mg, 0.652 mmol), and potassium carbonate (240 mg, 1.74 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (12.5 mL), and then tetra(triphenylphosphine)palladium (51 mg, 0.044 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the crude product was purified by thin-layer chromatography using silica gel plates (petroleum ether: ethyl acetate = 1:1) to obtain (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-b, 211 mg).

[0524] Step 3: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-c)

[0525] (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-b, 211 mg, 0.29 mmol) was dissolved in dichloromethane (3 mL), and then m-chloroperoxybenzoic acid (75 mg, 0.435 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to obtain crude product (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-c, 198 mg).

[0526] Step 4: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0527] Dissolve (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (129 mg, 0.81 mmol) in ultra-dry tetrahydrofuran (2 mL), add sodium hydrogen (13 mg, 0.54 mmol), stir at 0 °C for 30 minutes, then dissolve (1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (27-c, 198 mg, 198 ...((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (27-c, 198 mg, 198 mmol) in ultra-dry tetrahydrofuran (2 mL), add sodium hydrogen (13 mg, 0.54 mmol), stir at 0 °C for 30 minutes, then dissolve (((2 A solution of 0.27 mmol (mg, 0.27 mmol) dissolved in 3 mL of tetrahydrofuran was added. The reaction was carried out at room temperature for 1 hour. The reaction was quenched with water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel (dichloromethane:methanol = 10:1) to obtain the target product (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(27-d, 41 mg, yield 18.84%)). ESI[M+H +=806.4

[0528] Step 5: Synthesis of 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)-5-(((triisopropylsilyl)ethynyl)naphthalene-2-ol (27-e)

[0529] (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (27-d, 41 mg, 0.05 mmol)) was dissolved in acetonitrile (3 mL), and 1,4-dioxane hydrochloride solution (0.3 mL) was added. The reaction was carried out at room temperature for 1 hour. After concentration under reduced pressure, the crude product 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)-5-(((triisopropylsilyl)ethynyl)naphthyl-2-ol (27-e, 63 mg) was obtained. ESI[M+H] + =762.4

[0530] Step 6: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyrido[4,3-d]pyrimidin-7-yl)naphthalene-2-ol formate (27)

[0531] 6-Fluoro-4-(8-Fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)-5-(((triisopropylsilyl)ethynyl)naphthyl-2-ol (27-e, 63 mg, 0.083 mmol) was dissolved in N,N dimethylformamide (2 mL), and cesium fluoride (756 mg, 60.0 mmol) was added. The reaction was carried out at a warm temperature for 7 hours. Saturated brine and water were added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, the extract was purified by preparative liquid chromatography to obtain the target product, 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (27, 2.39 mg, yield 4.75%). ESI [M+H] + =606.2 1 H NMR(400MHz, DMSO-d6)δ 10.24(s, 0.5H), 8.20(s, 0.5H), 7.98-7.96(m, 1H), 7.48-746(m, 1H), 7.43-737(m, 1H), 7 .20-7.19(m,1H),5.34(s,0.5H),5.21(s,0.5H),5.20-4.71(m,1H),4.55-4.40(m,1H),4 .31-4.24(m, 2H), 4.13-3.98(m, 4H), 3.87(s, 3H), 3.61-3.58(m, 2H), 3.13-3.07(m, 2H), 3.01(s, 1H), 2.79-2.75(m, 2H), 2.12-2.11(m, 1H), 2.04-1.98(m, 2H), 1.86-1.77(m, 3H).

[0532] Example 28: Synthesis of 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphthalene-2-ol formate

[0533]

[0534] Step 1: Synthesis of 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyrido[4,3-d]pyrimidin-7-yl)naphthalene-2-ol formate (28)

[0535] 27.5 mg, 0.008 mmol of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (1 mL) was dissolved in tetrahydrofuran, and palladium on carbon (4 mg) was added. The reaction was carried out at room temperature for 1 hour under hydrogen protection. The mixture was then filtered with diatomaceous earth, the filter cake was washed with methanol, and the reaction solution was purified by preparative liquid chromatography to obtain the target product 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methoxypyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (28, 3.23 mg, yield 66.2%). ESI[M+H + =610.3 1 H NMR(400MHz, DMSO-d6)δ 10.22(s, 1H), 7.73(s, 1H), 7.33(s, 2H), 7.00-6.83(m, 1H), 5.30(s, 0.5H ), 5.17 (s, 0.5H), 4.86-4.81 (s, 1H), 4.56-4.44 (m, 1H), 4.43-4.38 (m, 2H) , 4.24-4.06(m, 3H), 3.98(s, 3H), 3.04-2.97(m, 3H), 2.97-2.77(m, 2H), 2. 08-2.00 (m, 3H), 1.81-1.72 (m, 3H), 1.42-1.20 (m, 4H), 0.82-0.76 (m, 3H).

[0536] Example 29: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-((((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0537]

[0538] Step 1: Synthesis of (2R,7aS)-7a-(chloromethyl)-2-fluorohexahydro-1H-pyrrolazine (29-A)

[0539] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (100 mg, 0.63 mmol) was dissolved in toluene (2.5 mL), and then thionyl chloride (0.068 mL, 0.945 mmol) was added. After reacting at 70 °C for 2.5 hours, the mixture was concentrated under reduced pressure to give the target product (2R,7aS)-7a-(chloromethyl)-2-fluorohexahydro-1H-pyrrolizin (29-A, 108 mg, yield 96.5%). ESI [M+H]+ = 178.1

[0540] Step 2: Synthesis of (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methylamine (29-B)

[0541] (2R,7aS)-7a-(chloromethyl)-2-fluorohexahydro-1H-pyrrolizine (29-A, 108 mg, 0.6 mmol) was dissolved in ammonia-methanol (5 mL), reacted at 70 °C for 16 hours, and then concentrated under reduced pressure to obtain the target product (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methylamine (29-B, 160 mg). ESI[M+H] + =159.1

[0542] Step 3: Synthesis of (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-a)

[0543] 7-Chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (2-a 496 mg, 1.8 mmol) was dissolved in acetonitrile (18 mL), followed by the addition of N,N-diisopropylethylamine (1.9 mL, 10.8 mmol) and phosphorus oxychloride (0.5 mL, 5.4 mmol). After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (1.9 mL, 10.8 mmol) and (R)-piperidin-3-ol (371.5 mg, 2.7 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–50%) to obtain the target product (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-a, 320 mg, yield 49.54%). ESI[M+H + =359.1

[0544] Step 4: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-b)

[0545] (R)-1-(7-chloro-8-fluoro-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-a, 143.5 mg, 0.4 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphthyl)ethynyl)triisopropylsilane (307.5 mg, 0.6 mmol), and potassium carbonate (221 mg, 1.6 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (5 mL), and then tetra(triphenylphosphine)palladium (46 mg, 0.04 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-b, 198 mg) was obtained by thin-layer chromatography with silica gel plates (petroleum ether: ethyl acetate = 3:1).

[0546] Step 5: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-c)

[0547] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-b, 198 mg, 0.28 mmol) was dissolved in dichloromethane (3 mL), and then m-chloroperoxybenzoic acid (73 mg, 0.42 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to obtain crude product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-c, 202 mg).

[0548] Step 6: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-d)

[0549] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methylamine (142 mg, 0.9 mmol) was dissolved in ultra-dry tetrahydrofuran (2 mL), and sodium hydrogen (14.5 mg, 0.6 mmol) was added. The mixture was stirred at 0 °C for 30 minutes. Then, a solution of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-c, 222 mg, 0.3 mmol) dissolved in tetrahydrofuran (2 mL) was added. The reaction was carried out at room temperature for 30 minutes. The product was quenched with water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyridano[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-d, 6 mg, yield 2.4%). ESI[M+H + =819.4

[0550] Step 7: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-e)

[0551] Dissolve ((R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-d, 6 mg, 0.007 mmol) in NN dimethylformamide (1 mL), and add cesium fluoride (8 mg, 0.042 mmol). The reaction was carried out at room temperature for 2 hours. Saturated brine was added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, the crude product (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-e, 10 mg). ESI [M+H]+ = 663.3

[0552] Step 8: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-((((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29)

[0553] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29-e, 10 mg, 0.015 mmol) was dissolved in acetonitrile (1 mL), and 1,4-dioxane hydrochloride solution (0.1 mL) was added. The reaction was carried out at room temperature for 2 hours. The pH was adjusted to 7 with a saturated sodium bicarbonate solution. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-((((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methyl)amino)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (29, 0.34 mg, yield 3.6%). ESI[M+H + =619.3 Example 30: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0554]

[0555] Step 1: Synthesis of 2-(chloromethyl)-1-methylpyrrolidine (30-A)

[0556] (1-Methylpyrrolidone-2-yl)methanol (600 mg, 5.209 mmol) was dissolved in toluene (15 mL), and thionyl chloride (1549 mg, 13.02 mmol) was added. The mixture was reacted at 70 °C for 3 hours. The reaction solution was concentrated under reduced pressure to give the target product, 2-(chloromethyl)-1-methylpyrrolidone (30-A, 1.1 g, yield 125%). ESI [M+H] + =169.05

[0557] Step 2: Synthesis of (1-methylpyrrolidone-2-yl)methylamine (30-B)

[0558] 2-(chloromethyl)-1-methylpyrrolidine (30-A, 1.1 g, 5.915 mmol) was dissolved in ammonia-methanol (12 ml) and reacted at 70 °C for 16 hours. The reaction solution was concentrated under reduced pressure to give the target product (1-methylpyrrolidine-2-yl)methylamine (30-B, 1 g, yield 134%). ESI [M+H] + =114.12

[0559] Step 3: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-b)

[0560] (1-Methylpyrrolidone-2-yl)methylamine (30-B, 400 mg, 3.505 mmol) was dissolved in tetrahydrofuran (2 ml) and N,N-dimethylformamide (2 ml), cooled to 0 °C, and sodium hydride (102 mg, 4.25 mmol) was added and reacted at 0 °C for 20 minutes. Then (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-a, 200 mg, 0.269 mmol) was added and the reaction was continued at 0 °C for 1 hour. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. After drying with anhydrous sodium sulfate, the solution was filtered, concentrated under reduced pressure, and purified by Pre-TLC (dichloromethane:methanol = 10:1) to obtain the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((((R)-1-methylpyrrolidone-2-yl)methyl)amino)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-b, 35 mg, yield 1.2%). ESI[M+H + =774.41

[0561] Step 4: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-c)

[0562] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-b, 35 mg, 0.045 mmol) was dissolved in N,N-dimethylformamide, cesium fluoride (100 mg, 0.658 mmol) was added, and the mixture was reacted at room temperature for 2 hours. Quenching with saturated NaCl aqueous solution, extraction with ethyl acetate, collection of the organic phase, drying with anhydrous sodium sulfate, filtration, and concentration under reduced pressure yielded the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-c, 30 mg, 111%). ESI[M+H + =618.28

[0563] Step 5: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30)

[0564] (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30-c, 30 mg, 0.0485 mmol) was dissolved in acetonitrile (0.5 ml), and dioxane hydrochloride (0.5 ml) was added. The reaction was carried out at room temperature for 0.5 hours. The pH was adjusted to 7 by adding sodium bicarbonate aqueous solution. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methoxy-2-((((R)-1-methylpyrrolidine-2-yl)methyl)amino)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (30, 0.64 mg, yield 2.3%). ESI [M+H] + =574.25, 1 HNMR (400MHz, DMSO-d6) δ10.12(s, 1H), 8.20(s, 1H), 7.96-7.83(m, 1H), 7.48-7.38(m, 1H), 7.32( s, 1H), 7.17-7.14 (m, 1H), 6.61 (s, 1H), 5.36-5.22 (m, 1H), 4.84 (s, 1H), 4.07-3.83 (m, 2H), 3.80 ( s, 1H), 3.71-3.51 (m, 2H), 3.39 (s, 2H), 3.14 (s, 3H), 2.94 (s, 2H), 2.64 (s, 1H), 2.32-2.30 (m, 1H) , 2.27-2.01(m, 2H), 2.01-1.90(m, 1H), 1.87-1.84(m, 1H), 1.71-1.30(m, 3H), 1.26-1.20(m, 2H).

[0565] Example 31: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol

[0566]

[0567] Step 1: Synthesis of 1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-a)

[0568] 2,4,7-Trichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidine (200 mg, 0.75 mmol) was dissolved in dichloromethane (10 mL) and cooled to 0 °C. N,N-diisopropylethylamine (194 mg, 1.50 mmol) and aziridine-3-ol hydrochloride (82 mg, 0.75 mmol) were added, and the mixture was reacted at 0 °C for 20 min. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column chromatography (methanol:dichloromethane = 0%–5%) to obtain the target product 1-(2,7-dichloro-8-fluoro-5-methylpyridino[4,3-d]pyrimidin-4-yl)aziridine-3-ol (31-a, 170 mg, yield 74.9%). ESI[M+H] + =266.4

[0569] Step 2: Synthesis of 1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-b)

[0570] 1-(2,7-dichloro-8-fluoro-5-methylpyridano[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-a, 170 mg, 0.56 mmol), ((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methanol (401 mg, 2.52 mmol), and N,N-diisopropylethylamine (183 mg, 1.42 mmol) were added to dioxane (8 ml). The mixture was heated to 90 °C and reacted for 16 h. Quenching with water, extraction with ethyl acetate, collection of the organic phase and repeated washing with saturated sodium chloride solution, drying with anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by Flash column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target product 1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-b, 140 mg, yield 58.8%). ESI[M+H + =303.1

[0571] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0572] 1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-b, 140 mg, 0.33 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)naphthyl)ethynyl)triisopropylsilane (202 mg, 0.39 mmol), cesium carbonate (323 mg, 0.99 mmol), and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (54 mg, 0.066 mmol) were dissolved in dioxane (7 ml) and water (1.75 ml). The reaction was purged with nitrogen three times and heated to 135℃ for 1 hour. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. After drying with anhydrous sodium sulfate, the mixture was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(31-c, 150 mg, yield 58.6%). ESI[M+H] + =776.2

[0573] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-d)

[0574] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(31-c, 150 mg, 0.193 mmol) was dissolved in acetonitrile (5 ml), and dioxane hydrochloride (1 ml) was added. The reaction was carried out at room temperature for 0.5 hours. The target product, 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-d, 97 mg, yield 80.1%), was obtained after purification. (ESI[M+H]) + =620.5

[0575] Step 5: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31)

[0576] 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31-d, 97 mg, 0.16 mmol) was dissolved in N,N-dimethylformamide (5 ml), and cesium fluoride (1.21 g, 8 mmol) was added. The mixture was reacted at room temperature for 4 hours. The reaction solution was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31.17 mg, yield 18.5%). ESI[M+H + =575.4, 1H NMR (400MHz, DMSO-d6) δ8.08 (s, 1H), 7.89-7.85 (m, 1H), 7.38-7.33 (m, 1H) 7.29-7.27 ( m, 1H), 7.12 (s, 1H), 5.66 (s, 1H), 5.13-5.12 (m, 0.5H), 5.11-5.10 (m, 0.5H), 4.66-4.60 (m, 1H), 4.52-4.44 (m, 1H), 4.25-4.20 (m, 2H), 4.15-4.01 (m, 1H), 3.96-3.90 (m, 2H), 3. 05-2.92(m, 4H), 2.78-2.69(m, 1H), 2.51(s, 3H), 2.05-1.89(m, 3H), 1.81-1.65(m, 3H).

[0577] Example 32: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol

[0578]

[0579] Step 1: Synthesis of 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (32)

[0580] 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (31.7 mg, 0.012 mmol) was dissolved in tetrahydrofuran (1 ml), 5 mg of palladium on carbon was added, hydrogen gas was introduced for purging, and the reaction was carried out at room temperature for 2 h. The mixture was then filtered through diatomaceous earth. The filtrate was purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)azacyclobutane-3-ol (32.2 mg, yield 28.7%). ESI [M+H] + =580.1, 1H NMR (400MHz, DMSO-d6) δ9.90 (s, 1H), 7.77-7.36 (m, 1H), 7.38-7.28 (m, 2H), 7.03-7 .00(m, 1H), 5.89-5.76(m, 1H), 5.37(s, 0.5H), 5.24(s, 0.5H), 4.66-4.60(m, 2H), 4 .24(s, 1H), 4.20-4.14(m, 1H), 4.12-4.04(m, 2H), 3.14(s, 1H), 2.62(s, 3H), 2.27- 2.14(m, 3H), 1.80-1.75(m, 3H), 1.40-1.31(m, 2H), 1.23(s, 4H), 0.78-0.74(m, 3H).

[0581] Example 33: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-4-(1,4-oxazin-4-yl)pyridino[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate

[0582]

[0583] Step 1: Synthesis of 4-(2,7-dichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidin-4-yl)-1,4-oxacyclopropane (33-a)

[0584] 2,4,7-Trichloro-8-fluoro-5-methyl-3,4-dihydropyrido[4,3-d]pyrimidine (150 mg, 0.56 mmol) was dissolved in dichloromethane (2 mL), followed by the addition of N,N-diisopropylethylamine (0.1 mL, 0.56 mmol) and 1,4-oxetine (57 mg, 0.56 mmol). The mixture was reacted at 0 °C for 0.5 hours, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel (petroleum ether:ethyl acetate = 1:1) to obtain the target product 4-(2,7-dichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidin-4-yl)-1,4-oxetine (33-a, 133 mg, yield 71.7%). ESI[M+H] + =331.0

[0585] Step 2: Synthesis of 4-(7-fluoro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-1,4-oxacyclopropane (33-b)

[0586] 4-(2,7-dichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidin-4-yl)-1,4-oxetane (33-a, 133 mg, 0.4 mmol) and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (287 mg, 1.8 mmol) were dissolved in 1,4-dioxane (7 mL), and then N,N-diisopropylethylamine (0.18 mL, 1.0 mmol) was added. The reaction was carried out at 90°C for 16 hours under nitrogen protection. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product 4-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-1,4-oxacyclopropane (33-b, 125 mg, yield 59.2%). ESI[M+H + =454.2

[0587] Step 3: Synthesis of 4-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0588] 4-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)-1,4-oxacyclopropane (33-b, 30 mg, 0.066 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3, 2-Dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (41 mg, 0.0792 mmol) and cesium carbonate (65 mg, 0.198 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (2.5 mL), followed by the addition of [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (11 mg, 0.0132 mmol). The mixture was reacted in a microwave oven at 135 °C for 1 hour. Water was added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, it was purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product 4-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(33-c, 22 mg, yield 44.45%). ESI[M+H] + =804.4

[0589] Step 4: Synthesis of 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-4-(1,4-oxacyclopentan-4-yl)pyridino[4,3-d]pyrimidin-7-yl)-5-((triisopropylsilyl)ethynyl)naphthalene-2-ol (33-d)

[0590] 4-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(33-c, 170 mg, 0.21 mmol) was dissolved in acetonitrile (3 mL), and 1,4-dioxane hydrochloride solution (0.2 mL) was added. The reaction was carried out at room temperature for 1 minute. Hours. After concentration under reduced pressure, the crude product 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-4-(1,4-oxacyclopentan-4-yl)pyridino[4,3-d]pyrimidin-7-yl)-5-((triisopropylsilyl)ethynyl)naphthyl-2-ol (33-d, 219 mg). ESI[M+H] + =760.4

[0591] Step 5: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-4-(1,4-oxetane-4-yl)pyrido[4,3-d]pyrimidin-7-yl)naphthalene-2-ol carboxylate (33)

[0592] 6-Fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-4-(1,4-oxacyclopentan-4-yl)pyrido[4,3-d]pyrimidin-7-yl)-5-((triisopropylsilyl)ethynyl)naphthyl-2-ol (33-d, 219 mg, 0.29 mmol) was dissolved in N,N dimethylformamide (3 mL), and cesium fluoride (2.2 g, 50.0 mmol) was added. The reaction was carried out at room temperature for 7 hours. The sample was extracted with ethyl acetate after adding saturated brine and water, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain the target product 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-4-(1,4-oxetane-4-yl)pyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (33, 15.45 mg, yield 8.82%). ESI[M+H +=604.3 1 H NMR(400MHz, DMSO-d6)δ 10.14(s, 1H), 7.93-7.91(m, 1H), 7.44-7.40(m, 1H), 7.35-7.20(m, 1H), 7.2 0-7.19 (m, 1H), 5.19 (s, 0.5H), 5.18 (s, 0.5H), 4.11-4.08 (m, 1H), 3.99-3.8 8(m, 1H), 3.86-3.70(m, 9H), 3.08-3.06(m, 2H), 2.99(s, 1H), 2.83-2.77(m, 1H), 2.56(s, 3H), 2.11-2.03(m, 1H), 2.00-1.96(m, 4H), 1.85-1.80(m, 3H).

[0593] Example 34: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol

[0594]

[0595] Step 1: Synthesis of (1-(2,7-dichloro-8-fluoro-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (34-a)

[0596] 2,4,7-Trichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidine (100 mg, 0.375 mmol) was dissolved in dichloromethane (5 mL) and cooled to 0 °C. N,N-diisopropylethylamine (96 mg, 0.75 mmol) and aziridine-3-ylmethanol hydrochloride (48 mg, 0.375 mmol) were added, and the mixture was reacted at 0 °C for 20 min. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column chromatography (methanol:dichloromethane = 0%–5%) to obtain the target product (1-(2,7-dichloro-8-fluoro-5-methylpyridino[4,3-d]pyrimidine-4-yl)aziridine-3-yl)methanol (34-a, 107 mg, yield 89.9%). ESI[M+H) + =317.1

[0597] Step 2: Synthesis of 1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (34-b)

[0598] 1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (34-a, 105 mg, 0.33 mmol), ((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methanol (264 mg, 1.66 mmol), and N,N-diisopropylethylamine (128 mg, 0.99 mmol) were added to dioxane (5 ml). The mixture was heated to 90 °C and reacted for 16 h. Quenching with water, extraction with ethyl acetate, collection of the organic phase and repeated washing with saturated sodium chloride solution, drying with anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by Flash column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target product (1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (34-b, 84 mg, yield 57.5%). ESI[M+H + =439.9

[0599] Step 3: Synthesis of (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0600] (1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)azacyclobutane-3-yl)methanol (34-b, 84 mg, 0.19 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (117 mg, 0.23 mmol), cesium carbonate (247 mg, 0.76 mmol), and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (31 mg, 0.038 mmol) were dissolved in dioxane (4 ml) and water (1 ml). The reaction was purged with nitrogen three times and heated to 135℃ for 1 hour. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. After drying with anhydrous sodium sulfate, the mixture was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target product (1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(34-c, 98 mg, yield 65.3%)). ESI[M+H + =790.0

[0601] Step 4: Synthesis of 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)-5-(((triisopropylsilyl)ethynyl)naphthalene-2-ol) (34-d)

[0602] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(34-c, 98 mg, 0.124 mmol) was dissolved in acetonitrile (5 ml), and dioxane hydrochloride (1 ml) was added. The reaction was carried out at room temperature for 0.5 hours. The reaction solution was purified. The target product, 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)-5-(((triisopropylsilyl)ethynyl)naphthyl-2-ol (34-d, 61 mg, yield 65.6%), was obtained by ESI[M+H]. +=746.1

[0603] Step 5: Synthesis of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (34)

[0604] Dissolve 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)-5-(((triisopropylsilyl)ethynyl)naphthyl-2-ol (34-d, 61 mg, 0.08 mmol) in N,N-dimethylformamide (3 ml), and add cesium fluoride (600 mg). The reaction mixture (g, 4 mmol) was reacted at room temperature for 4 hours. The reaction solution was purified by preparative liquid chromatography to obtain the target product 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (34, 20 mg, yield 42.5%). ESI [M+H] + =589.9, 1 H NMR (400MHz, DMSO-d6) δ8.21 (s, 1H), 7.96-7.94 (m, 1H), 7.49-7.35 (m, 2H), 7.22(s, 1H), 5.32(s, 0.5H), 5.21(s, 0.5H), 4.55-4.50(m, 1H), 4.32-4.14(m , 3H), 4.14-4.09(m, 1H), 4.04-4.01(m, 1H), 3.97-3.93(m, 2H), 3.16-3.00(m , 4H), 2.88-2.80(m, 2H), 2.59(s, 3H), 2.20-1.93(m, 4H), 1.87-1.74(m, 3H).

[0605] Example 35: Synthesis of 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol

[0606]

[0607] Step 1: Synthesis of 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (35)

[0608] 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (34.7 mg, 0.012 mmol) was dissolved in tetrahydrofuran (1 ml), 5 mg of palladium on carbon was added, hydrogen gas was purged, and the reaction was carried out at room temperature for 2 h. The mixture was then filtered through diatomaceous earth. The filtrate was purified by preparative liquid chromatography to obtain the target product, 5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(3-(hydroxymethyl)azacyclobutane-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-ol (35.2 mg, yield 28.6%). ESI[M+H + =594.2, 1 H NMR (600MHz, DMSO-d6) δ9.90 (s, 1H), 7.77-7.73 (m, 1H), 7.33-7.30 (m, 2H), 7.03 (s, 1H), 5 .36(s, 0.5H), 5.22(s, 0.5H), 4.49-4.45(m, 1H), 4.24-4.14(m, 2H), 4.08-4.01(m, 2H), 3. 60-3.57(m, 2H), 3.12-3.02(m, 2H), 2.86-9-2.82(mz, 2H), 2.62(s, 2H), 2.18-2.14(m, 2H) , 2.07-2.01(m, 2H), 1.89-1.77(m, 3H), 1.35(s, 3H), 1.26-1.16(m, 2H), 0.78-0.74(m, 3H).

[0609] Example 36: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol

[0610]

[0611] Step 1: Synthesis of (R)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-a)

[0612] 2,4,7-Trichloro-8-fluoro-5-methyl-3,4-dihydropyrido[4,3-d]pyrimidine (50 mg, 0.19 mmol) was dissolved in dichloromethane (1 mL), followed by the addition of N,N-diisopropylethylamine (0.033 mL, 0.19 mmol) and (R)-pyrrolidine-3-ol (16.5 mg, 0.19 mmol). The mixture was reacted at 0 °C for 0.25 hours, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel (petroleum ether:ethyl acetate = 1:1) to obtain the target product (R)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidine-4-yl)pyrrolidine-3-ol (36-a, 58 mg, yield 94.3%). ESI[M+H] + =317.0

[0613] Step 2: Synthesis of (R)-1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-b)

[0614] (R)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-a, 58 mg, 0.18 mmol) and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (129 mg, 0.81 mmol) were dissolved in 1,4-dioxane (2 mL), and then N,N-diisopropylethylamine (0.078 mL, 0.45 mmol) was added. The reaction was carried out at 90°C for 16 hours under nitrogen protection. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product (R)-1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-b, 46 mg, yield 68.0%). ESI[M+H + =440.2

[0615] Step 3: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0616] (R)-1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-b, 46 mg, 0.1 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3-yl)pyrrolidine-3-ol) 2-Dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (61.5 mg, 0.12 mmol) and cesium carbonate (98 mg, 0.3 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (2.5 mL), and then [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (16 mg, 0.02 mmol) was added. The mixture was reacted in a microwave at 135 °C for 1 hour. Water was added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, it was purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(36-c, 48 mg, yield 49.8%). ESI[M+H] + =790.4

[0617] Step 4: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-d)

[0618] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(36-c, 89 mg, 0.11 mmol) was dissolved in acetonitrile (3 mL), and 1,4-dioxane hydrochloride solution (1 mL) was added. The reaction was carried out at room temperature for 1 minute. Hours. Concentration under reduced pressure yielded the crude product (R)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-d, 79 mg). ESI[M+H] + =746.4

[0619] Step 5: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36)

[0620] (R)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36-d, 79 mg, 0.1 mmol) was dissolved in N,N-dimethylformamide (3 mL), and cesium fluoride (911.4 mg, 60.0 mmol) was added. The reaction was carried out at room temperature for 12 hours. The sample was extracted with ethyl acetate after adding saturated brine and water, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (36, 6.62 mg, yield 11.2%). ESI[M+H + =590.2 1 H NMR(400MHz, DMSO-d6)δ 10.12-10.09(m,1H),7.96-7.91(m,1H),7.44-7.39(m,1H),7.36-7.34(m,1H),7.26 -714(m, 1H), 5.33(s, 0.5H), 5.32(s, 0.5H), 5.20-4.92(m, 1H), 4.33-4.21(m, 1H), 4. 20-4.08 (m, 2H), 4.04-3.96 (m, 2H), 3.85-3.65 (m, 2H), 3.42-3.40 (m, 1H), 3.15-3.01 (m, 3H), 2.84-2.79 (m, 1H), 2.54-2.52 (m, 3H), 2.12-1.98 (m, 4H), 1.95-1.78 (m, 4H).

[0621] Example 37: Synthesis of (S)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol carboxylate

[0622]

[0623] Step 1: Synthesis of (S)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-a)

[0624] 2,4,7-Trichloro-8-fluoro-5-methyl-3,4-dihydropyrido[4,3-d]pyrimidine (50 mg, 0.19 mmol) was dissolved in dichloromethane (1 mL), followed by the addition of N,N-diisopropylethylamine (0.033 mL, 0.19 mmol) and (S)-pyrrolidine-3-ol (16.5 mg, 0.19 mmol). The mixture was reacted at 0 °C for 0.5 hours, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel (petroleum ether:ethyl acetate = 1:1) to obtain the target product (S)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidine-4-yl)pyrrolidine-3-ol (37-a, 51 mg, yield 84.6%). ESI[M+H] + =317.0

[0625] Step 2: Synthesis of (S)-1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-b)

[0626] Dissolve (S)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-a, 51 mg, 0.16 mmol) and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (203 mg, 1.28 mmol) in 1,4-dioxane (2 mL), and then add N,N-diisopropylethylamine (0.111 mL, 0.64 mmol). The reaction was carried out at 90°C for 16 hours under nitrogen protection. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product (S)-1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-b, 53 mg, yield 75.3%). ESI[M+H + =440.2

[0627] Step 3: Synthesis of (S)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0628] (S)-1-(7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-b, 53 mg, 0.12 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3-yl)pyrrolidine-3-ol) 2-Dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (74 mg, 0.144 mmol) and cesium carbonate (117 mg, 0.36 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (2.5 mL), and then [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (20 mg, 0.024 mmol) was added. The mixture was reacted in a microwave at 135 °C for 1 hour. Water was added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, it was purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product (S)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(37-c, 41 mg, yield 43.2%). ESI[M+H] + =790.4

[0629] Step 4: Synthesis of (S)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-d)

[0630] (S)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(37-c, 41 mg, 0.05 mmol) was dissolved in acetonitrile (2 mL), and 1,4-dioxane hydrochloride solution (1 mL) was added. The reaction was carried out at room temperature for 1 minute. Hours. After concentration under reduced pressure, the crude product (S)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-d, 51 mg). ESI[M+H] + =746.4

[0631] Step 5: Synthesis of (S)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol carboxylate (37)

[0632] (S)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37-d, 51 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (2 mL), and cesium fluoride (638 mg, 60.0 mmol) was added. The reaction was carried out at room temperature for 8 hours. The sample was extracted with ethyl acetate after adding saturated brine and water, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain the target product (S)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (37, 3.35 mg, yield 8.1%). ESI[M+H + =590.2 1H NMR(400MHz, DMSO-d6)δ 10.12(s, 1H), 8.12-7.96(m, 1H), 7.44-7.42(m, 1H), 7.39-7.34(m, 1H), 7.25-713( m, 1H), 5.33 (s, 0.5H), 5.32 (s, 0.5H), 5.19-5.04 (m, 1H), 4.35-4.33 (m, 1H), 4.32- 4.13(m, 2H), 4.11-3.99(m, 2H), 3.94-3.77(m, 2H), 3.46-3.43(m, 1H), 3.09-3.00( m, 3H), 2.82-2.79 (m, 1H), 2.54-2.52 (m, 3H), 2.12-1.97 (m, 4H), 1.88-1.74 (m, 4H).

[0633] Example 38: (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0634]

[0635] Step 1: (R)-1-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)piperidin-3-ol (38)

[0636] Referring to Example 37, (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)piperidin-3-ol (38-a) was prepared.

[0637] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-5-methylpyridin[4,3-d]pyrimidin-4-yl)piperidin-3-ol (38-a, 81 mg, 0.1 mmol) was dissolved in N,N-dimethylformamide (2 mL), and cesium fluoride (638 mg, 1 mmol) was added. The reaction was carried out at room temperature for 8 hours. Saturated brine and water were added, and the mixture was extracted with ethyl acetate. After concentration under reduced pressure, the extract was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)pyrrolidine-3-ol (38, 30.1 mg, yield 51%), ESI [M+H). + =648.

[0638] Example 39: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0639]

[0640] Step 1: Synthesis of (R)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-a)

[0641] 2,4,7-Trichloro-8-fluoro-5-methylpyridinone[4,3-d]pyrimidine (50 mg, 0.188 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. N,N-diisopropylethylamine (48 mg, 0.376 mmol) and (R)-piperidin-3-ol hydrochloride (26 mg, 0.188 mmol) were added, and the mixture was reacted at 0 °C for 20 min. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column chromatography (methanol:dichloromethane = 0%–5%) to obtain the target product (R)-1-(2,7-dichloro-8-fluoro-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-a, 59 mg, yield 95.0%). ESI[M+H] + =331.1

[0642] Step 2: Synthesis of (R)-1-(7-chloro-8-fluoro-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-b)

[0643] (R)-1-(2,7-dichloro-8-fluoro-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-a, 59 mg, 0.18 mmol), (S)-(1-methylpyrrolidone-2-yl)methanol (103 mg, 0.89 mmol), and N,N-diisopropylethylamine (70 mg, 0.54 mmol) were added to dioxane (3 ml). The mixture was heated to 90 °C and reacted for 16 h. Quenching with water, extraction with ethyl acetate, collection of the organic phase and repeated washing with saturated sodium chloride solution, drying with anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by Flash column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target product (R)-1-(7-chloro-8-fluoro-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-b, 60 mg, yield 81.2%). ESI[M+H + =409.8

[0644] Step 3: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy]pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-c)

[0645] (R)-1-(7-chloro-8-fluoro-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-b, 60 mg, 0.146 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)naphthyl)ethynyl)triisopropylsilane (90 mg, 0.176 mmol), cesium carbonate (143 mg, 0.438 mmol), and 1,1′-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (24 mg, 0.029 mmol) were dissolved in dioxane (3 ml) and water (0.75 ml). The reaction was purged with nitrogen three times and heated to 135℃ for 1 hour. The mixture was quenched with water, extracted with ethyl acetate, and the organic phase was collected and repeatedly washed with saturated sodium chloride solution. After drying with anhydrous sodium sulfate, the mixture was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (methanol:dichloromethane = 0%–10%) to obtain the target product (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy]pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-c, 34 mg, yield 31.0%). ESI[M+H + =760.0

[0646] Step 4: Synthesis of (R)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-d)

[0647] (R)-1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy]pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-c, 34 mg, 0.045 mmol) was dissolved in acetonitrile (2.5 ml), and dioxane hydrochloride (0. 5 ml), reacted at room temperature for 0.5 hours. The reaction solution was purified to obtain the target product (R)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-d, 20 mg, yield 62.5%). ESI[M+H + =716.1

[0648] Step 5: Synthesis of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39)

[0649] (R)-1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39-d, 20 mg, 0.028 mmol) was dissolved in N,N-dimethylformamide (1 ml), and cesium fluoride (212 mg, 1.4 mmol) was added. The mixture was reacted at room temperature for 4 hours. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-5-methyl-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (39.2 mg, yield 33.3%). ESI[M+H + =560.2, 1 H NMR (400MHz, DMSO-d6) δ8.30 (s, 1H), 8.08-7.82 (m, 1H), 7.51-7.36 (m, 1H), 7.32-7 .17(m, 1H), 6.64(s, 1H), 5.33-5.31(m, 1H), 4.47-4.30(m, 1H), 4.29-4.21(m, 1H), 3.89-3.78(m, 3H), 2.99-2.87(m, 2H), 2.77-2.57(m, 2H), 2.36-2.34(m, 2H), 2.16- 2.12(m, 2H), 2.01-1.97(m, 2H), 1.75-1.42(m, 3H), 1.37-1.10(m, 6H), 0.85(s, 1H).

[0650] Example 40: Preparation of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl dimethylcarbamate trifluoroacetate

[0651]

[0652] Step 1: Preparation of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl dimethylcarbamate trifluoroacetate (40)

[0653] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (4,100 mg, 0.166 mmol) was dissolved in dichloromethane (5 ml), and dimethylcarbamoyl chloride (17.85 mg, 0.166 mmol) and dimethylaminopyridine (20 mg, 0.166 mmol) were added at room temperature, and the reaction was carried out for 2 h. The reaction solution was subjected to vacuum distillation to remove the solvent. The crude product was purified by preparative liquid chromatography (aqueous phase: 0.01% TFA, organic phase: acetonitrile) to obtain the target product 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl dimethylcarbamate trifluoroacetate (40, 36.7 mg, yield 28.02%). ESI[M+H] + =675.40. 1 H NMR (400MHz, DMSO-d6) δ10.92-10.84 (d, 1H), 8.21-8.17 (m, 1H), 7.99-7.98 (t, 1H), 7.67-7.44 (m, 2H), 5.6 4(s, 0.5H), 5.51(s, 0.5H), 4.65-4.55(m, 2H), 4.35-4.32(d, 1H), 4.07(s, 4H), 4.03-3.96(m, 2H), 3.88-3.8 1(m,3H),3.78(s,2H),3.47-3.37(d,1H),3.11-3.10(d,3H),2.94-2.91(d,3H),2.67-2.62(d,3H),2.36-2 .29(m, 1H), 2.24-2.12(m, 2H), 2.07-2.01(m, 1H), 1.87-1.75(m, 2H), 1.71-1.60(m, 1H), 1.55-1.47(m, 1H).

[0654] Example 41: Preparation of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate (41)

[0655]

[0656] Step 1: Preparation of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate (41)

[0657] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (4,100 mg, 0.166 mmol) was dissolved in dichloromethane (5 ml), and acetic anhydride (37.28 mg, 0.365 mmol), triethylamine (50 mg, 0.498 mmol) and dimethylaminopyridine (4 mg, 0.033 mmol) were added at room temperature, and the reaction was carried out for 2 h. The reaction solution was subjected to vacuum distillation to remove the solvent. The crude product was purified by preparative liquid chromatography (aqueous phase: 0.05% NH4HCO3, organic phase: acetonitrile) to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridano[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate (41, 32.15 mg, yield 30%). ESI[M+H] + =646.3. 1HNMR (400MHz, DMSO-d6) δ10.10-10.08 (d, 1H), 7.98-7.92 (m, 1H), 7.45-7.40 (t, 1H), 7.35 (s, 1H), 7.26-7.11 (m, 1H), 5.34 (s, 0.5H), 5.20 (s, 0.5H), 4.94-4.82 (d, 1H), 4.16 (s, 1H) , 4.05-4.02(d, 1H), 3.88-3.80(m, 2H), 3.67-3.53(m, 2H), 3.14-3.02(m, 3H), 2.86-2.80( m, 1H), 2.67-2.63 (m, 3H), 2.13 (s, 1H), 2.06-1.93 (m, 5H), 1.88-1.73 (m, 6H), 1.66 (s, 2H).

[0658] Example 42: Preparation of trifluoroacetate of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate

[0659]

[0660] Step 1: Preparation of trifluoroacetate of (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate (42)

[0661] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate (41, 28.83 mg, 0.045 mmol) was dissolved in dichloromethane (5 ml), and acetic anhydride (5 mg, 0.05 mmol), triethylamine (14 mg, 0.135 mmol) and dimethylaminopyridine (5 mg, 0.045 mmol) were added at room temperature, and the reaction was carried out for 2 h. The reaction solution was subjected to vacuum distillation to remove the solvent. The crude product was purified by preparative liquid chromatography (aqueous phase: 0.05% NH4HCO3, organic phase: acetonitrile) to obtain the target product (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidin-3-yl acetate trifluoroacetate (42.25 mg, yield 69.44%). + =688.4. 1 HNMR (400MHz, DMSO-d6) δ10.78 (s, 1H), 8.21-8.17 (m, 1H), 7.97-7.96 (d, 1H), 7.64-7.57 (t, 1H) ,7.58-7.43(m,1H),5.61(s,1H),5.49(s,1H),4.94-4.84(d,1H),4.67-4.53(m,2H),3.98-3.93( m, 1H), 3.90-3.79 (m, 2H), 3.78-3.66 (m, 3H), 3.59-3.57 (m, 2H), 3.29-3.14 (m, 3H), 2.69 (s, 3H) , 2.53-2.48(d, 1H), 2.34-2.25(m, 4H), 2.22-2.10(m, 2H), 2.05-1.89(m, 4H), 1.83-1.58(m, 3H).

[0662] Example 43: Preparation of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl 4-methylbenzenesulfonate trifluoroacetate

[0663]

[0664] Step 1: Preparation of trifluoroacetate of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl 4-methylbenzenesulfonate (43)

[0665] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (4,118 mg, 0.2 mmol) was dissolved in dichloromethane (5 ml), and triethylamine (61 mg, 0.6 mmol) and p-toluenesulfonyl chloride (76 mg, 0.4 mmol) were added at room temperature, and the reaction was allowed to proceed for 10 minutes. The reaction solution was subjected to vacuum distillation to remove the solvent, and the crude product was purified by preparative liquid chromatography (aqueous phase: 0.01% TFA, organic phase: acetonitrile) to obtain the target product 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4, [3-d]pyrimidin-7-yl)naphthyl-2-yl4-methylbenzenesulfonate trifluoroacetate (43, 118.31 mg, yield 67.85%). ESI [M+H]+=758.1.1H NMR (400MHz, DMSO-d6) δ 10.98–10.91 (d, 1H), 8.25–8.21 (m, 1H), 8.00–7.97 (m, 1H) ,7.87-7.76(m,2H),7.69-7.64(m,1H),7.50-7.48(d,2H),7.38-7.28(m,1H),5.64 (s, 0.5H), 5.51 (s, 0.5H), 4.66-4.56 (m, 2H), 4.36-4.33 (d, 1H), 4.09-3.97 (m, 4H), 3.91-3.79(m, 6H), 3.48-3.39(m, 1H), 2.68-2.53(m, 4H), 2.42(s, 3H), 2.34-2.29( m, 1H), 2.23-2.20 (m, 2H), 2.12-1.95 (m, 2H), 1.85-1.83 (m, 1H), 1.68-1.45 (m, 2H).

[0666] Example 44: Preparation of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidine-3-ol carboxylate

[0667]

[0668] Step 1: Preparation of (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidine-3-ol carboxylate (44)

[0669] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol carboxylate (4,15 mg, 0.02 mmol) was dissolved in tetrahydrofuran (2 ml), and 15 mg of 5% palladium on carbon was added at room temperature. After reacting for 16 hours, the mixture was filtered, and the solvent was removed by vacuum distillation to obtain the target product (R)-1-(7-(8-ethyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol carboxylate (44 mg, 10.3 mg, yield 78.81%). ESI[M+H + =608.4. 1HNMR (400MHz, DMSO-d6) δ9.93-9.95 (d, 1H), 7.78-7.73 (m, 1H), 7.39-7.28 (m, 2H), 7.09-6.99 (m, 1H), 5.37 (s, 0.5H), 5. 20(s, 0.5H), 5.12-4.78(d, 1H), 4.27-4.24(t, 1H), 4.22-4.12(m, 1H), 4.04-4.07(d, 1H), 3.83-3.77(m, 1H), 3.70-3.56 (m, 1H), 3.13 (s, 2H), 3.06 (s, 1H), 2.86-2.84 (m, 1H), 2.64 (s, 3H), 2.44-2.40 (t, 1H), 2.25-2.21 (m, 1H), 2.14-2.10 (m, 2H), 2.08-2.02(m, 1H), 2.01-1.98(m, 2H), 1.92-1.82(m, 3H), 1.80-1.71(m, 3H), 1.52-1.43(m, 1H), 0.80-0.67(m, 3H).

[0670] Example 45: Preparation of 5-acetylene: 6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl acetate trifluoroacetate

[0671]

[0672] Step 1: Preparation of trifluoroacetate of 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl acetate (45)

[0673] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (4.60 mg, 0.1 mmol) was dissolved in dichloromethane (6 ml), and triethylamine (40 mg, 0.4 mmol) and acetyl chloride (7.85 mg, 0.1 mmol) were added at 0 °C, and the reaction was continued for 10 minutes. The reaction solution was subjected to vacuum distillation to remove the solvent. The crude product was purified by preparative liquid chromatography (aqueous phase: 0.01% TFA, organic phase: acetonitrile) to obtain the target product 5-ethynyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl acetate trifluoroacetate (45, 50.49 mg, yield 66.43%). ESI[M+H] + =646.5. 1 HNMR (400MHz, DMSO-d6) δ11.01-11.10 (d, 1H), 8.23-8.19 (m, 1H), 8.00-7.98 (t, 1H), 7.68-7.49 (m, 2H), 5. 63(s, 0.5H), 5.50(s, 0.5H), 4.65-4.56(m, 2H), 4.39-4.29(m, 1H), 4.09(s, 1H), 4.02-3.95(m, 2H), 3.86-3. 82 (m, 3H), 3.79-3.75 (m, 2H), 3.50-3.25 (m, 3H), 2.67 (s, 1H), 2.59-2.54 (m, 2H), 2.48-2.42 (m, 1H), 2.36-2 .29(m, 4H), 2.23-2.16(m, 2H), 2.10-2.01(m, 1H), 1.92-1.80(m, 1H), 1.74-1.58(m, 1H), 1.57-1.49(m, 1H).

[0674] Example 46: Preparation of trifluoroacetate of (5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl)ethyl carbonate

[0675]

[0676] Step 1: Preparation of trifluoroacetate of (5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl)carbonate (46)

[0677] (R)-1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (4,60 mg, 0.1 mmol) was dissolved in dichloromethane (6 ml), and triethylamine (30 mg, 0.3 mmol) and diethyl dicarbonate (19.5 mg, 0.12 mmol) were added at room temperature, and the reaction was continued for 10 minutes. The reaction solution was subjected to vacuum distillation to remove the solvent. The crude product was purified by preparative liquid chromatography (aqueous phase: 0.01% TFA, organic phase: acetonitrile) to obtain the target product (5-ethyl-6-fluoro-4-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4-(((R)-3-hydroxypiperidin-1-yl)-5-methylpyridino[4,3-d]pyrimidin-7-yl)naphth-2-yl)ethyl carbonate trifluoroacetate (46,45 mg, yield 56.96%). + =676.2. 1 HNMR (400MHz, DMSO-d6) δ11.04-10.94 (d, 1H), 8.22-8.18 (m, 1H), 8.10-8.08 (t, 1H), 7.72-7.58 (m, 2H), 5.61 (s, 0.5H) , 5.48 (s, 0.5H), 4.62-4.53 (m, 2H), 4.34-4.25 (m, 3H), 4.12-4.09 (m, 2H), 4.01-3.94 (m, 2H), 3.84 (s, 1H), 3.81-3.79 ( d, 2H), 3.76-3.67 (m, 2H), 3.49-3.29 (m, 2H), 2.67-2.61 (m, 1H), 2.58-2.51 (m, 2H), 2.45-2.44 (m, 1H), 2.31-2.26 (m, 1 H), 2.19-2.10(m, 2H), 2.07-1.94(m, 2H), 1.83-1.77(m, 1H), 1.70-1.54(m, 1H), 1.52-1.44(m, 1H), 1.31-1.27(m, 3H).

[0678] Example 47: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol

[0679]

[0680] Step 1: Synthesis of 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-a)

[0681] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1, 1.68 g, 6.0 mmol) was dissolved in acetonitrile (30 mL), followed by the addition of N,N-diisopropylethylamine (6.3 mL, 36 mmol) and phosphorus oxychloride (1.7 mL, 18 mmol). After reacting at 80 °C for 1 hour, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (6.3 mL, 36 mmol) and 3-methylazacyclobutane-3-ol (1.1 g, 9.0 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 0%–600%) to obtain the target product 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-a, 765 mg, yield 31.4%). ESI [M+H]+ = 349.0

[0682] Step 2: Synthesis of 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-b)

[0683] 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-a, 453 mg, 1.3 mmol), methylboronic acid (156 mg, 2.6 mmol), and potassium phosphate (827 mg, 3.9 mmol) were dissolved in a 10:1 mixture of toluene and water (13.2 mL), and then dichloro[1,1′-bis(diphenylphosphine)ferrocene]palladium(II) (190 mg, 0.26 mmol) was added. The reaction was carried out at 105 °C for 16 hours under nitrogen protection. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (petroleum ether:ethyl acetate = 1:1) to obtain the target product 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-b, 128 mg, yield 29.9%). ESI [M+H] + =329.1

[0684] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-c)

[0685] 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-b, 160 mg, 0.49 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (377 mg, 0.735 mmol), and potassium carbonate (270 mg, 1.96 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (6 mL), and then tetra(triphenylphosphine)palladium (57 mg, 0.049 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (petroleum ether: ethyl acetate = 1:1) to obtain the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-c, 164 mg).

[0686] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-d)

[0687] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-c, 164 mg, 0.24 mmol) was dissolved in dichloromethane (3 mL), and then m-chloroperoxybenzoic acid (62 mg, 0.36 mmol) was added. The reaction was carried out at room temperature for 1 hour. Add saturated sodium bicarbonate solution, extract with dichloromethane, concentrate under reduced pressure to give crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-d, 198 mg)

[0688] Step 5: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (47-e)

[0689] ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (134 mg, 0.84 mmol) was dissolved in ultra-dry tetrahydrofuran (2 mL), and sodium hydrogen (13 mg, 0.56 mmol) was added. The mixture was stirred at 0 °C for 30 minutes. Then, a solution of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-d, 198 mg, 0.28 mmol) dissolved in tetrahydrofuran (2 mL) was added. The reaction was carried out at room temperature for 30 minutes. The sample was quenched with water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel plates (dichloromethane:methanol = 10:1) to obtain the target product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(47-e, 46 mg, yield 20.8%). ESI[M+H] + =790.4

[0690] Step 6: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-f)

[0691] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(47-e, 46 mg, 0.06 mmol) was dissolved in acetonitrile (2 mL), and 1,4-dioxane hydrochloride solution (0.2 mL) was added. The reaction was carried out at room temperature for 1 hour. After concentration, the crude product was obtained as 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-f, 59 mg). ESI[M+H] + =746.4

[0692] Step 7: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47)

[0693] 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47-f, 59 mg, 0.08 mmol) was dissolved in N,N-dimethylformamide (2 mL), and cesium fluoride (729 mg, 60.0 mmol) was added. The reaction was carried out at room temperature for 4 hours. The sample was extracted with ethyl acetate after adding saturated brine and water, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain the target product 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)-3-methylazacyclobutane-3-ol (47, 5.58 mg, yield 11.83%). ESI [M+H] + =590.2 1 H NMR (400MHz, DMSO-d6)δ 10.08(s, 1H), 7.95-7.91(m, 1H), 7.42-7.39(m, 1H), 7.35-7.34(m, 1H), 7.17(s, 1H), 5.32(s, 0.5H), 5.18(s, 0.5H), 4.34-4.31(m, 2H), 4.29-4.1 39(m, 4H), 3.86(s, 1H), 3.10-3.00(m, 3H), 2.87-2.79(m, 1H), 2.59(s, 3H ), 2.11-2.03(m, 1H), 2.06-1.95(m, 2H), 1.87-1.71(m, 3H), 1.38(s, 3H).

[0694] Example 48: Synthesis of (3R)-1-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-7-(5-methyl-1H-indazol-4-yl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0695]

[0696] Step 1: Synthesis of (R)-1-(5,7-di-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-a)

[0697] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidin[4,3-d]pyrimidin-4(3H)-one (I-1, 600 mg, 2.14 mmol) was dissolved in acetonitrile (30 ml), and then phosphorus oxychloride (986 mg, 6.43 mmol) and N,N-diisopropylethylamine (1.68 g, 13 mmol) were added. After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (840 mg, 1.605 mmol) and (R)-piperidine-3-ol hydrochloride (324 mg, 2.14 mmol) were added. The reaction was then carried out at 0 °C for 0.5 hours. The solution was quenched with water, extracted with ethyl acetate, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered, concentrated under reduced pressure, and purified by Flash column chromatography (ethyl acetate: petroleum ether = 30%–35%) to obtain the target product (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-a, 430 mg, yield 52.3%). ESI [M+H] + =363.2

[0698] Step 2: Synthesis of (R)-1-(7-chloro-5-methyl-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-b)

[0699] (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-a, 430 mg, 1.14 mmol) was dissolved in toluene (20 ml) and water (2 ml), followed by the addition of methylboronic acid (341 mg, 5.7 mol), potassium phosphate (726 mg, 3.42 mmol), and 1,1′-bis(diphenylphosphino)ferrocene palladium(II) dichloride (167 mg, 0.23 mmol). The mixture was heated to 105 °C and reacted for 16 h. After concentration under reduced pressure, the product was purified by TLC (petroleum ether:ethyl acetate = 1:1) to obtain the target product (R)-1-(7-chloro-5-methyl-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-b, 230 mg, yield 56.5%). ESI[M+H] + =343.1

[0700] Step 3: Synthesis of (3R)-1-(8-fluoro-5-methyl-7-(5-methyl-1H-indazol-4-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-c)

[0701] (R)-1-(7-chloro-5-methyl-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-b, 230 mg, 0.72 mmol), potassium carbonate (400 mg, 2.88 mmol), tetrakis(triphenylphosphine)palladium (166 mg, 0.144 mmol), (2-fluoro-6-(methoxymethyloxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (553 mg, 1.08 mmol) were dissolved in 1,4-dioxane (10 ml) and water (2.5 ml), purged with nitrogen for 1 minute, and reacted in a microwave at 135 °C for 1 hour. TLC (petroleum ether:ethyl acetate = 1:1) following vacuum concentration yielded the target product (3R)-1-(8-fluoro-5-methyl-7-(5-methyl-1H-indazol-4-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-c, 450 mg, yield 88.4%). ESI [M+H] + =438.5

[0702] Step 4: Synthesis of (3R)-1-(8-fluoro-5-methyl-7-(5-methyl-1H-indazol-4-yl)-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-d)

[0703] (3R)-1-(8-fluoro-5-methyl-7-(5-methyl-1H-indazol-4-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-c, 450 mg, 0.64 mmol) and m-chloroperoxybenzoic acid (228 mg, 0.95 mmol) were dissolved in dichloromethane (8 ml). The reaction was carried out at room temperature for 0.5 h. The reaction was quenched with water, extracted with dichloromethane, and the organic phase was collected and washed with water in saturated sodium bicarbonate solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the target product (3R)-1-(8-fluoro-5-methyl-7-(5-methyl-1H-indazol-4-yl)-2-(methylsulfinyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-d, 423 mg, yield 96.8%). ESI[M+H] + =454.4

[0704] Step 5: Synthesis of (3R)-1-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-7-(5-methyl-1H-indazol-4-yl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48)

[0705] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrololin-7a(5H)-yl)methanol (282 mg, 1.77 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C and 60% sodium hydride (47 mg, 1.17 mmol) was added, and the reaction was continued for 0.5 hours. Then (3R)-1-(8-fluoro-5-methyl-7-(5-methyl-1H-indazol-4-yl)-2-(methylsulfinyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48-d, 423 mg, 0 ml) was added. 0.59 mmol), reacted for 0.5 h. Quenched with water, extracted with ethyl acetate, the organic phase was collected and washed with water in saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the target product (3R)-1-(8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-7-(5-methyl-1H-indazol-4-yl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (48,298 mg, yield 22.9%). ESI[M+H + =550.1, 1 H NMR (400MHz, DMSO-d6) δ8.38 (s, 1H), 7.82-7.49 (m, 1H), 7.39-7.11 (m, 2H), 5.33-5. 31(m, 0.5H), 5.22-5.20(m, 0.5H), 4.14-4.11(m, 1H), 4.09-3.98(m, 2H), 3.78(s, 1H ), 3.14-3.06(m, 2H), 3.04-2.79(m, 2H), 2.70-2.67(m, 2H), 2.30(s, 3H), 2.17-1.95 (m, 4H), 1.93-1.66 (m, 3H), 1.47-1.44 (m, 2H), 1.27-1.23 (m, 4H), 1.19-1.15 (m, 2H).

[0706] Example 49: Synthesis of (R)-1-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol

[0707]

[0708] Step 1: Synthesis of (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-a)

[0709] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrimidine[4,3-d]pyrimidine-4(3H)-one (I-1, 600 mg, 2.14 mmol) was dissolved in acetonitrile (30 ml), and then phosphorus oxychloride (986 mg, 6.43 mmol) and N,N-diisopropylethylamine (1.68 g, 13 mmol) were added. After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (828 mg, 6.43 mmol) and (R)-piperidine-3-ol hydrochloride (885 mg, 6.43 mmol) were added. The reaction was then carried out at 0 °C for 0.5 hours. Quenching with water, extraction with ethyl acetate, collection of the organic phase, washing with water in saturated sodium bicarbonate solution, drying with anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by Flash column chromatography (ethyl acetate: petroleum ether = 30%–35%) to obtain the target product (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-a, 400 mg, yield 51.5%). ESI [M+H + =363.2

[0710] Step 2: Synthesis of (R)-1-(7-chloro-5-cyclopropyl-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-b)

[0711] (R)-1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-a, 400 mg, 1.1 mmol) was dissolved in toluene (4 ml) and water (0.4 ml), and then cyclopropylboronic acid (114 mg, 1.32 mol), potassium phosphate (700 mg, 3.3 mmol), palladium acetate (50 mg, 0.22 mmol), and 2-biscyclohexylphosphine-2,6-dimethoxy-diphenyl (90 mg, 0.22 mmol) were added. The mixture was heated to 105 °C and reacted under nitrogen protection for 16 h. After concentration under reduced pressure, TLC (petroleum ether:ethyl acetate = 1:1) purification yielded the target product (R)-1-(7-chloro-5-cyclopropyl-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-b, 198 mg, yield 48.8%). ESI [M+H] + =368.8

[0712] Step 3: Synthesis of (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-c)

[0713] (R)-1-(7-chloro-5-cyclopropyl-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-b, 198 mg, 0.54 mmol), potassium carbonate (299 mg, 2.15 mmol), tetrakis(triphenylphosphine)palladium (139 mg, 0.12 mmol), (2-fluoro-6-(methoxymethyloxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (332 mg, 0.65 mmol) were dissolved in 1,4-dioxane (10 ml) and water (2 ml), purged with nitrogen for 1 minute, and reacted in a microwave at 135 °C for 1 hour. After concentration under reduced pressure, TLC (petroleum ether:ethyl acetate = 1:1) yielded the target product (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthyl-1-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-c, 201 mg, yield 51.8%). ESI [M+H] + =718.8

[0714] Step 4: Synthesis of (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-d)

[0715] (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-c, 201 mg, 0.28 mmol) and m-chloroperoxybenzoic acid (72 mg, 0.42 mmol) were dissolved in dichloromethane (8 ml). The reaction was carried out at room temperature for 0.5 h. The solution was quenched with water, extracted with dichloromethane, and the organic phase was collected and washed with water in a saturated sodium bicarbonate solution. After drying with anhydrous sodium sulfate, the solution was filtered and concentrated under reduced pressure to obtain the target product (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(methanesulfonyl)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-d, 178 mg, yield 86.4%). ESI[M+H + =734.9

[0716] Step 5: Synthesis of (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (49-e)

[0717] (((2R,7aS)-2-fluorotetrahydro-1H-pyrrolino-7a(5H)-yl)methanol (111 mg, 0.73 mmol) was dissolved in anhydrous tetrahydrofuran (5 ml), the temperature was lowered to 0 °C and 60% sodium hydride (29 mg, 0.73 mmol) was added, and the reaction was continued for 0.5 hours. Then (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-d, 178 mg) was added. 0.24 mmol), reacted for 0.5 h. Quenched with water, extracted with ethyl acetate, the organic phase was collected and washed with water in saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and TLC (DCM∶MeOH=10∶1) yielded the target product (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(49-e, 89 mg, yield 44.7%). ESI[M+H] + =830.4

[0718] Step 6: Synthesis of (R)-1-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy]pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-f)

[0719] (R)-1-(5-cyclopropyl-8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(49-e, 89 mg, 0.11 mmol) was dissolved in N,N-dimethylformamide (3 ml), and cesium fluoride (167 mg, 1.1 mmol) was added. The reaction was carried out at 25 °C for 1 hour. Water was added to the reaction solution. Extracted three times with ethyl acetate (20 ml each time), the organic phase was collected and distilled under reduced pressure to give the target product (R)-1-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy]pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-f, 46 mg, yield 62.2%). ESI[M+H + =673.7

[0720] Step 7: Synthesis of (R)-1-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49)

[0721] (R)-1-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-(methoxy-methoxy)naphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy]pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49-f, 46 mg, 0.07 mmol) was dissolved in acetonitrile (2.5 ml), and 1,4-dioxane hydrochloride solution was added at 25 °C. (0.5 ml), reaction time 1 h. The reaction solution was purified by preparative liquid chromatography to obtain the target product (R)-1-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-ol (49, 15 mg, yield 34.1%). ESI[M+H + =630.1, 1H NMR (400MHz, DMSO-d6) δ9.96-9.91 (m, 1H), 8.00-7.90 (m, 1H), 7.47-7.41 (m, 1H), 7 .33-7.30(m, 1H), 7.04-6.90(m, 1H), 5.40(s, 0.5H), 5.26(s, 0.5H), 4.25-4.10(m, 4H), 3.52(s, 1H), 3.24-3.02(m, 3H), 2.89(s, 1H), 2.45-2.31(m, 2H), 2.29-1.99(m , 4H), 1.91-1.83(m, 4H), 1.51-1.44(m, 4H), 1.16-1.12(m, 2H), 1.02-0.96(m, 3H).

[0722] Example 50: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate

[0723]

[0724] Step 1: Synthesis of 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-a)

[0725] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (I-1, 196 mg, 0.7 mmol) was dissolved in acetonitrile (5 mL), followed by the addition of N,N-diisopropylethylamine (2.6 mL, 4.2 mmol) and phosphorus oxychloride (0.2 mL, 2.1 mmol). After reacting at 80 °C for 0.5 hours, the reaction was cooled to 0 °C, and then N,N-diisopropylethylamine (2.6 mL, 4.2 mmol) and piperidine-3-carbamate (154 mg, 0.05 mmol) were added, followed by a reaction at 0 °C for 1 hour. The solution was quenched in water, extracted with ethyl acetate, concentrated under reduced pressure, and purified by thin-layer chromatography using silica gel plates (petroleum ether:ethyl acetate = 3:1) to obtain the target product 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-carbamate (50-a, 130 mg, yield 49.9%). (ESI[M+H) + =372.0

[0726] Step 2: Synthesis of 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-b)

[0727] 1-(5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-a, 30 mg, 0.08 mmol), methylboronic acid (7 mg, 0.12 mmol), and potassium phosphate (68 mg, 0.32 mmol) were dissolved in a 10:1 mixture of toluene and water (1.1 mL), and then dichloro[1,1′-bis(diphenylphosphine)ferrocene]palladium(II) (12 mg, 0.016 mmol) was added. The reaction was carried out at 105 °C for 16 hours under nitrogen protection. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel (petroleum ether:ethyl acetate = 2:1) to obtain the target product 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-b, 4 mg, yield 14.2%). ESI[M+H] + =352.1

[0728] Step 3: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-c)

[0729] 1-(7-chloro-8-fluoro-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidin-3-carbamate (50-b, 35 mg, 0.09 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (69 mg, 0.135 mmol), and potassium carbonate (50 mg, 0.36 mmol) were dissolved in a 4:1 mixture of 1,4-dioxane and water (2.5 mL), and then tetra(triphenylphosphine)palladium (10 mg, 0.009 mmol) was added. The mixture was microwaved at 135 °C for 1 hour. After concentration under reduced pressure, the product was purified by thin-layer chromatography using silica gel plates (petroleum ether:ethyl acetate = 1:1) to obtain the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-c, 30 mg, yield 47.5%). ESI[M+H] + =702.3

[0730] Step 4: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-d)

[0731] 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methylthio)pyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-c, 30 mg, 0.04 mmol) was dissolved in dichloromethane (1 mL), and then m-chloroperoxybenzoic acid (10.4 mg, 0.06 mmol) was added. The reaction was carried out at room temperature for 1 hour. The sample was extracted with dichloromethane after adding a saturated sodium bicarbonate solution and concentrated under reduced pressure to give the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-d, 28 mg, yield 95.5%). ESI[M+H + =734.3

[0732] Step 5: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)

[0733] Dissolve ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (18 mg, 0.114 mmol) in ultra-dry tetrahydrofuran (1 mL), add sodium hydrogen (3 mg, 0.076 mmol), stir at 0 °C for 30 minutes, then add a solution of 1-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-5-methyl-2-(methanesulfonyl)pyridino[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-d, 30 mg, 0.038 mmol) dissolved in tetrahydrofuran (1 mL). React at room temperature for 30 minutes. The sample was quenched with water, extracted with ethyl acetate, and concentrated under reduced pressure to give the crude product 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy(50-e, 26 mg, yield 84.2%). ESI[M+H] + =813.4

[0734] Step 6: Synthesis of 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyrido[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-f)

[0735] 1-(8-fluoro-7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy (50-e, 26 mg, 0.03 mmol) was dissolved in acetonitrile (3 mL), and 1,4-dioxane hydrochloride solution (0.5 mL) was added. The reaction was carried out at room temperature for 1 hour. At that time, after concentration under reduced pressure, crude product was obtained: 1-(8-fluoro-7-(7-fluoro-3-hydroxy-8-((triisopropylsilyl)ethynyl)naphth-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methylpyridino[4,3-d]pyrimidin-4-yl)piperidine-3-carbamate (50-f, 27 mg). ESI[M+H] + =769.4

[0736] Step 7: Synthesis of 1-(7-(8-ethynyl-7-fluoro-3-hydroxynaphth-1-yl)-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin...

Claims

1. The following compounds or their pharmaceutically acceptable salts: 。 2. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a medicament for treating cancer or immune diseases.

4. The use according to claim 3, wherein, The cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, bile duct cancer, endometrial cancer, or ovarian cancer.

5. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a medicament for treating diseases associated with KRAS mutations.

6. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a KRAS inhibitor.

7. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of KRAS G12D, KRAS G12V, KRAS G12A, KRAS G12S, KRAS G12C, KRAS G13D, KRAS Q61H, and KRAS Q61K mutation inhibitors.

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