KRAS G12C protein mutation inhibitors, methods of making, pharmaceutical compositions, and uses thereof

Abstract

The application discloses a KRAS G12C protein mutation inhibitor, a preparation method, a pharmaceutical composition and application thereof. And specifically discloses a compound as shown in formula I, a solvate, a prodrug, a metabolite or a pharmaceutically acceptable salt thereof, the compound has good KRAS G12C protein mutation inhibition activity, and can be used for treating or preventing cancer.

Description

Technical Field

[0001] This invention relates to a KRAS G12C Protein mutation inhibitors, their preparation methods, pharmaceutical compositions and their applications. Background Technology

[0002] RAS (Rat sarcoma) proteins, including KRAS, HRAS, and NRAS, are members of the small GTPase family. RAS proteins can bind to either guanine trinucleotide phosphate (GTP) or guanine dinucleotide phosphate (GDP), with the switching between these states regulated by guanine nucleotide exchange factors (GEFs) and GTP-activated protein activator (GAP). When bound to GDP, RAS proteins are in an "inactive" state; when bound to GTP, they are in an "activated" state. In the GTP-bound state, RAS proteins directly interact with various downstream effector proteins and activate signaling pathways, including mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K), thereby regulating cell growth, cell motility, metabolism, and the interaction between the cell and its extracellular environment.

[0003] Overactivation mutations in the RAS gene are the most common gene mutations in cancer, with approximately 30% of human tumors carrying some form of mutated RAS gene. KRAS is the most common mutated oncogene, accounting for 86% of all RAS mutations. Most RAS gene mutations reduce the binding of GAP to RAS proteins, thus affecting GAP's ability to catalyze GTP hydrolysis. This leads to a decrease in the intrinsic GTPase activity of the RAS protein, or both, resulting in an increase in GTP-bound RAS protein, keeping it in a persistently "activated" state. Activated RAS protein continuously activates downstream signaling pathways, ultimately leading to sustained cell growth and differentiation, and consequently, tumorigenesis. RAS mutations are a genetic driver of many cancer types, including colorectal cancer (CRC), pancreatic ductal adenocarcinoma (PDAC), lung adenocarcinoma (LUAD; a subtype of non-small cell lung cancer (NSCLC)), melanoma, and certain hematologic malignancies.

[0004] In KRAS-mutated tumors, the majority of oncogenic mutations occur within codon 12, accounting for 83%. The G12C mutation is a relatively common subtype of KRAS gene mutation. G12C Mutations are prevalent in lung adenocarcinoma and also found in other tumor types such as pancreatic and colorectal cancer. Despite extensive drug discovery efforts, there are currently no drugs targeting KRAS. G12C Marketed drugs with mutations. KRAS G12C Mutations in KRAS have been shown to be associated with poor cancer prognosis. Therefore, targeting KRAS...G12C The development of mutation inhibitors is of significant clinical need. Shokat and colleagues reported a covalently related KRAS... G12C Inhibitors are used to treat KRAS G12C Mutated tumors offer possibilities (Ostrem JM., et al. Nature, 2013, 548-551). Although a number of patents have been published regarding KRAS G12C inhibitors (WO2014143659, WO2014152588, WO2015054572, WO2016049524, WO2017201161, WO2018119183, etc.), it remains very important to discover more effective KRAS inhibitors with better pharmacokinetic and pharmacodynamic properties. Summary of the Invention

[0005] The technical problem to be solved by this invention is that it addresses the shortcomings of existing KRAS. G12C The limitation of protein mutation inhibitors having relatively simple structures leads to the development of a KRAS-based approach. G12C Protein mutation inhibitors, their preparation methods, pharmaceutical compositions and their applications: The compounds of this invention have novel structures and good activity and selectivity.

[0006] This invention provides a compound of Formula I, its solvate, its prodrug, its metabolite, or a pharmaceutically acceptable salt thereof (referring to the aforementioned compound of Formula I, its solvate, its prodrug, or its metabolite):

[0007]

[0008] Ring A is a 4-12 membered alicyclic ring containing 2-4 N atoms; the alicyclic ring is a monocyclic, bridged, or spirocyclic ring.

[0009] R 1 -C(=O)-C(R) a )=C(R b (R) c -C(=O)-C≡CR c -C(=O)-C(=O)-OR d or -C(=O)-R d ;

[0010] R a It is hydrogen, halogen or C 1-6 alkyl;

[0011] R b and R c Independently hydrogen, C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 alkyl";

[0012] Each R b-1 Independent of halogen, C 1-6 Alkoxy or -NR b-2 R b-3 ;R b-2 and R b-3 Independently hydrogen or C 1-6 Alkyl, or R b-2 R b-3 Together with the N atoms connected to them, they form "4-10 membered heterocyclic alkyl groups with N atoms, or N and O atoms, and one or two heteroatoms";

[0013] R d C 1-6 alkyl;

[0014] n is 0, 1, 2, or 3;

[0015] Each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 alkyl";

[0016] Each R 4-1 Independently halogen, cyano, hydroxyl or C 1-6 Alkoxy;

[0017] R 2 For -OR 2-1 -SR 2-2 -C(=O)R 2-3 -S(=O)2R 2-4 -S(=O)R 2-5 、-(CR 1-1 R 1-2 ) m R 2-6 or NR 1- 3 R 2-7 ;

[0018] R 1-1 R 1-2 and R 1-3 Independently hydrogen or C 1-6 alkyl;

[0019] m can be 0, 1, 2, or 3. When m is 0, it represents R. 2-6 It is directly connected to the parent body via a single bond;

[0020] R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5Independently for C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group with one or more heteroatoms selected from N, O and S, and having 1-4 heteroatoms", and is surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl;

[0021] R 2-6 Hydrogen, CN, halogen, hydroxyl, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group with one or more heteroatoms selected from N, O and S, and having 1-4 heteroatoms", and is surrounded by one or more R e -1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl;

[0022] R 2-7 For hydrogen, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6alkoxy, "a 5-12 membered heteroaryl group with one or more heteroatoms selected from N, O and S, and having 1-4 heteroatoms", and is surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl;

[0023] Each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e -4 Replacement C 3-10 cycloalkyl;

[0024] R e-1 R e-2 and R e-4 Independent of halogen, C 1-6 Alkyl, C 1-6 Alkoxy group, -C(=O)-C(R) a )=C(R b (R) c ) or NR e-5 R e-6 ;R e-5 and R e-6 Independently hydrogen or C 1-6 Alkyl, or R e-5 R e-6Together with the N atoms connected to them, they form "4-10 membered heterocyclic alkyl groups with N atoms, or N and O atoms, and one or two heteroatoms";

[0025] Each R e-3 Independently halogen, hydroxyl, cyano, C 1-6 Alkyl, with one or more R e-7 Replacement C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R e-8 Replacement C 1-6 Alkoxy group, -C(=O)-C(R) a )=C(R b (R) c ), NR e-5 R e-6 -O(C=O)NR e-13 R e-14 or -O(C=O)OR e-15 ;R e-7 and R e-8 Independently halogen, hydroxyl, cyano, -COOH, C 1-6 Alkoxy or NR e -9 R e-10 R e-9 and R e-10 Independently hydrogen, C 1-6 Alkyl, -C(=O)R e-11 or -S(=O)2R e-12 ;R e-11 and R e-12 Independently for C 1-6 alkyl;

[0026] R e-13 and R e-14 Independently hydrogen or C 1-6 Alkyl, or R e-13 R e-14 Together with the N atoms connected to them, they form "4-10 membered heterocyclic alkyl groups with N as the heteroatom, or N and O, and one or two heteroatoms"; R e-15 It is hydrogen or C 1-6 alkyl;

[0027] R 3 For hydrogen, C 1-6 Alkyl, with one or more R 3-1 Replacement C 1-6 Alkyl groups, wherein each R 3-1 Independently for C 1-6 Alkyl, C 1-6 Alkoxy, CN, halogen, or OH;

[0028] X and Y are independently O and NR. 6 or CR 7 R 8 Each R 6 Independently hydrogen or C 1-6 Alkyl groups; each R 7 and R 8 Independently H or C 1-6 Alkyl; Z represents a single bond, O, or NR 6 or CR 7 R 8 X, Y, and Z are not all 0 or not all NR at the same time. 6 ;

[0029] U, V, W, and Q are independently N or CR 5 ;

[0030] Each R 5 Independently H, halogen, hydroxyl group, or C substituted with one or more halogens. 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkoxy, cyano, C 3-6 cycloalkyl, C 1-6 Alkyl sulfone, C 3-6 cycloalkylsulfone group, C 1-6 Alkyl or C 1-6 Alkoxy;

[0031] Carbon atoms marked with an asterisk (*) are chiral carbon atoms, and are in the S configuration, R configuration, or a mixture thereof.

[0032] In certain preferred embodiments of the present invention, certain groups in the compounds represented by Formula I, their solvates, their prodrugs, their metabolites, or their pharmaceutically acceptable salts are defined as follows, and groups not mentioned are as described in any embodiment of the present invention (hereinafter referred to as "in one embodiment of the present invention").

[0033] Ring A is a 4-12 membered alicyclic ring containing 2-4 N atoms; the alicyclic ring is a monocyclic, bridged, or spirocyclic ring.

[0034] R 1 -C(=O)-C(R) a )=C(R b (R) c -C(=O)-C≡CR c -C(=O)-C(=O)-OR d or -C(=O)-R d ;

[0035] R a It is hydrogen, halogen or C 1-6 alkyl;

[0036] R b and R c Independently hydrogen, C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 alkyl";

[0037] Each R b-1 Independent of halogen, C 1-6 Alkoxy or -NR b-2 R b-3 ;R b-2 and R b-3 Independently hydrogen or C 1-6 Alkyl, or R b-2 R b-3 Together with the N atoms connected to them, they form "4-10 membered heterocyclic alkyl groups with N atoms, or N and O atoms, and one or two heteroatoms";

[0038] R d C 1-6 alkyl;

[0039] n is 0, 1, 2, or 3;

[0040] Each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 alkyl";

[0041] Each R 4-1 Independently halogen, cyano, hydroxyl or C 1-6 Alkoxy;

[0042] R 2 For -OR 2-1 -SR 2-2 -C(=O)R 2-3 -S(=O)2R 2-4 -S(=O)R 2-5 、-(CR 1-1 R 1-2 ) m R 2-6 or NR 1- 3 R 2-7 ;

[0043] R 1-1 R 1-2 and R 1-3 Independently hydrogen or C 1-6 alkyl;

[0044] m can be 0, 1, 2, or 3. When m is 0, it represents R. 2-6It is directly connected to the parent body via a single bond;

[0045] R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group with one or more heteroatoms selected from N, O and S, and having 1-4 heteroatoms", and is surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl;

[0046] R 2-6 Hydrogen, CN, halogen, hydroxyl, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group with one or more heteroatoms selected from N, O and S, and having 1-4 heteroatoms", and is surrounded by one or more R e -1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl;

[0047] R 2-7 For hydrogen, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group with one or more heteroatoms selected from N, O and S, and having 1-4 heteroatoms", and is surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl;

[0048] Each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", C 3-10 cycloalkyl, or, by one or more R e -4 Replacement C 3-10 cycloalkyl;

[0049] R e-1 R e-2 and R e-4 Independent of halogen, C 1-6 Alkyl, C 1-6 Alkoxy group, -C(=O)-C(R) a )=C(R b (R) c ) or NR e-5 Re-6 ;R e-5 and R e-6 Independently hydrogen or C 1-6 Alkyl, or R e-5 R e-6 Together with the N atoms connected to them, they form "4-10 membered heterocyclic alkyl groups with N atoms, or N and O atoms, and one or two heteroatoms";

[0050] Each R e-3 Independently halogen, hydroxyl, cyano, C 1-6 Alkyl, with one or more R e-7 Replacement C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R e-8 Replacement C 1-6 Alkoxy group, -C(=O)-C(R) a )=C(R b (R) c ), NR e-5 R e-6 -O(C=O)NR e-13 R e-14 or -O(C=O)OR e-15 ;R e-7 and R e-8 Independently halogen, hydroxyl, cyano, -COOH, C 1-6 Alkoxy or NR e -9 R e-10 R e-9 and R e-10 Independently hydrogen, C 1-6 Alkyl, -C(=O)R e-11 or -S(=O)2R e-12 ;R e-11 and R e-12 Independently for C 1-6 alkyl;

[0051] R e-13 and R e-14 Independently hydrogen or C 1-6 Alkyl, or R e-13 R e-14 Together with the N atoms connected to them, they form "4-10 membered heterocyclic alkyl groups with N as the heteroatom, or N and O, and one or two heteroatoms"; R e-15 It is hydrogen or C 1-6 alkyl;

[0052] R 3 For hydrogen, C 1-6 Alkyl, with one or more R 3-1 Replacement C1-6 Alkyl groups, wherein each R 3-1 Independently for C 1-6 Alkyl, C 1-6 Alkoxy, CN, halogen, or OH;

[0053] X and Y are independently O and NR. 6 Or CH2; R 6 It is hydrogen or C 1-6 Alkyl; Z represents a single bond, O, or NR 6 Or CH2; X, Y, and Z are not simultaneously O or not simultaneously NR 6 ;

[0054] U, V, W, and Q are independently N or CR 5 ;

[0055] Each R 5 Independently H, halogen, hydroxyl group, or C substituted with one or more halogens. 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkoxy, cyano, C 3-6 cycloalkyl, C 1-6 Alkyl sulfone, C 3-6 cycloalkylsulfone group, C 1-6 Alkyl or C 1-6 Alkoxy;

[0056] Carbon atoms marked with an asterisk (*) are chiral carbon atoms, and are in the S configuration, R configuration, or a mixture thereof.

[0057] In one aspect of the present invention, when ring A is a 4-12 membered alicyclic ring containing 2-4 N atoms, the alicyclic ring is a 6-10 membered alicyclic ring containing 2 N atoms, for example... for

[0058] In one aspect of the present invention, when R a When the halogen is halogen, the halogen is fluorine, chlorine, bromine or iodine, such as fluorine.

[0059] In one aspect of the present invention, when R a C 1-6 When alkyl, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0060] In one aspect of the present invention, when R b and R c Independently for C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl is a C1-C3 alkyl, such as methyl.

[0061] In one aspect of the present invention, when each R b-1 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine, or iodine, such as fluorine.

[0062] In one aspect of the present invention, when each R b-1 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group, such as a methoxy group.

[0063] In one aspect of the present invention, when R b-2 and R b-3 Independently for C 1-6 When alkyl, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0064] In one aspect of the present invention, when R d C 1-6 When alkyl, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0065] In one aspect of the present invention, when each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl is a C1-C3 alkyl, such as methyl.

[0066] In one aspect of the present invention, when each R 4-1 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine, or iodine, such as fluorine.

[0067] In one aspect of the present invention, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 Alkyl or "with one or more R e Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl is a C1-C3 alkyl, such as methyl.

[0068] In one aspect of the present invention, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group, such as a methoxy group.

[0069] In one aspect of the present invention, when R 2-1 R2-2 R 2-3 R 2-4 and R 2-5 Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a heteroaryl group containing one or more R atoms". e-1 When replacing "a 5-12-membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and whose heteroatoms number 1-4", the phrase "a 5-12-membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and whose heteroatoms number 1-4" refers to...

[0070] In one aspect of the present invention, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently defined as "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms" or "a group consisting of one or more R..." e-3 When replacing "a heteroatom selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms", the "heteroatoms selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms" is...

[0071] In one aspect of the present invention, when R 2-6 When the halogen is halogen, the halogen is fluorine, chlorine, bromine or iodine, such as chlorine.

[0072] In one aspect of the present invention, when R 2-6 C 1-6 Alkyl or "with one or more R e Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl is a C1-C3 alkyl, such as methyl.

[0073] In one aspect of the present invention, when R 2-6 C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group, such as a methoxy group.

[0074] In one aspect of the present invention, when R 2-6 Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a heteroaryl group containing one or more R atoms". e-1 When replacing "a 5-12-membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and whose heteroatoms number 1-4", the phrase "a 5-12-membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and whose heteroatoms number 1-4" refers to...

[0075] In one aspect of the present invention, when R 2-6 "A 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms" or "a group consisting of one or more R..." e-3 When replacing "a heteroatom selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms", the "heteroatoms selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms" is...

[0076] In one aspect of the present invention, when each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a heteroaryl group containing one or more R atoms". e-1 When replacing "a 5-12-membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and whose heteroatoms number 1-4", the phrase "a 5-12-membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and whose heteroatoms number 1-4" refers to...

[0077] In one aspect of the present invention, when each R e Independently defined as "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms" or "a group consisting of one or more R..." e-3 The substituted "heteroatom selected from one or more of O and N, and having 1-3 heteroatoms in a 4-10 membered heterocyclic alkyl group" is... For example

[0078] In one aspect of the present invention, when R e-1 R e-2 and R e-4 When independently a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0079] In one aspect of the present invention, R e-3 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine, or iodine, such as fluorine.

[0080] In one aspect of the present invention, R e-3 Independently C1-C6 alkyl or "containing one or more R e-7 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl is a C1-C3 alkyl, such as methyl.

[0081] In one aspect of the present invention, R e-3 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group, such as a methoxy group.

[0082] In one aspect of the present invention, when R e-5 and R e-6 When independently a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl or ethyl.

[0083] In one aspect of the present invention, when R 3 When it is a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0084] In one aspect of the present invention, when each R 5 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine or iodine, for example fluorine, chlorine or bromine, or more specifically fluorine or chlorine.

[0085] In one aspect of the present invention, when each R 5 C that is independently replaced by one or more halogens 1-6 Alkyl, the C substituted with one or more halogens 1-6 Alkyl groups are C atoms substituted with one or more halogens. 1-3 Alkyl groups, such as -CF3.

[0086] In one aspect of the present invention, when each R 5 Independently for C 1-6 When alkyl, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0087] In one aspect of the present invention, when each R 5 C that is independently replaced by one or more halogens 1-6 Alkoxy, the C substituted with one or more halogens 1-6 The alkoxy group is a C-aryl group substituted with one or more halogens. 1-3 Alkyl groups, for example, -OCF3.

[0088] In one aspect of the present invention, when each R 7 and R 8 Independently for C 1-6 When alkyl, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl.

[0089] In one embodiment of the present invention, ring A is a 6-10 membered alicyclic ring containing 2 N atoms, and the alicyclic ring is a monocyclic ring.

[0090] In one aspect of the present invention, R 1 -C(=O)-C(R)a )=C(R b (R) c ), -C(=O)-C≡CCH3, -C(=O)-C(=O)-OCH3 or -C(=O)-CH3; when R a Halogen or C 1-6 When alkyl, R b and R c It is hydrogen; when R b or R c C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 When "alkyl", R a It is hydrogen.

[0091] In one aspect of the present invention, R a It is hydrogen or halogen.

[0092] In one aspect of the present invention, R b and R c Independently hydrogen, C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 "alkyl", and R b and R c At least one of them is hydrogen.

[0093] In one embodiment of the present invention, each R b-1 Halogens are independent of each other.

[0094] In one aspect of the present invention, R b and R c Independently hydrogen or "C substituted with one or more halogens" 1-6 "alkyl", and R b and R c At least one of them is hydrogen.

[0095] In one embodiment of the present invention, n is 0, 1 or 2; preferably 1 or 2.

[0096] In one embodiment of the present invention, each R 4 The methyl group is independently methyl, cyano-substituted methyl, F-substituted methyl, or hydroxy-substituted methyl; preferably cyano-substituted methyl.

[0097] In one aspect of the present invention, R 2-1 For hydrogen, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl or "with one or more R e-1The substituted heteroatom is "a 5-12-membered heteroaryl group selected from one or more of N, O and S, and having 1-4 heteroatoms".

[0098] In one aspect of the present invention, R 2-2 C 1-6 Alkyl or "with one or more R e Replacement C 1-6 alkyl".

[0099] In one aspect of the present invention, R 2-5 For one or more R e Replacement C 1-6 alkyl.

[0100] In one embodiment of the present invention, m is 0.

[0101] In one aspect of the present invention, R 2-6 Hydrogen, halogen, or one or more R e-3 The substituted "4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", "5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", or "substituted by one or more R..." e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms or a hydroxyl group.

[0102] In one embodiment of the present invention, each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted "5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms", "4-10-membered heterocycloalkyl group selected from one or more of O and N, with 1-3 heteroatoms", or "substituted by one or more R..." e-3 The substituted heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms; preferably, each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a heteroaryl group containing one or more R atoms". e-3 The substituted heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms.

[0103] In one aspect of the present invention, R e-1 Independently for C 1-6 alkyl.

[0104] In one embodiment of the present invention, each Re-3 Independently halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy group, -C(=O)-C(R) a )=C(R b (R) c ) or NR e-5 R e-6 Preferably, R a R b and R c All are H.

[0105] In one aspect of the present invention, R 2 For -OR 2-1 -SR 2-2 -S(=O)R 2-5 or -(CR) 1-1 R 1-2 ) m R 2-6 .

[0106] In one aspect of the present invention, R 3 It is hydrogen or C 1-6 alkyl.

[0107] In one embodiment of the present invention, X is O or CR. 7 R 8 Y is CR 7 R 8 Z represents a single bond, O, or CR. 7 R 8 .

[0108] In one embodiment of the present invention, X and Y are CH2, and Z is a single bond, O, or CH2.

[0109] In one embodiment of the present invention, U, V, W, and Q are independently CR. 5 .

[0110] In one embodiment of the present invention, each R 5 Independently, H, halogen, or C substituted with one or more halogens. 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkoxy, cyano, C 3-6 cycloalkyl, C 1-6 Alkyl or C 1-6 Alkyl group.

[0111] In one embodiment of the present invention, each R 5 Independent of H, halogen or C 1-6 alkyl.

[0112] In one aspect of the present invention, for

[0113] In one aspect of the present invention, R 1 for

[0114] In one aspect of the present invention, R 2 For -OR 2-1 or -(CR) 1-1 R 1-2 ) m R 2-6 R 2-1 For one or more R e Replacement C 1-6 Alkyl groups, each R e Independently for one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", R e In the middle, each R e-3 Independent of halogen, C 1-6 Alkyl or C 1-6 Alkoxy group; m is 0, R 2-6 For one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", R 2-6 In the middle, each R e-3 Independent for NR e-5 R e-6 .

[0115] In one aspect of the present invention, for

[0116] In one aspect of the present invention, R 2 For hydrogen, hydroxyl, chlorine, -OCH3, -SCH3

[0117] In one aspect of the present invention, for

[0118] Preferably, the compound represented by Formula I is any of the following compounds:

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128] Preferably, the compound represented by Formula I is any of the following compounds:

[0129] The compound with a retention time of 1.21 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0130] The compound with a retention time of 1.99 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0131] The compound with a retention time of 0.93 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0132] The compound with a retention time of 1.43 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0133] The compound with a retention time of 1.24 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0134] The compound with a retention time of 2.56 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0135] The compound with a retention time of 1.01 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0136] The compound with a retention time of 2.17 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0137] The compound with a retention time of 4.74 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0138] The compound with a retention time of 5.44 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0139] The compound with a retention time of 4.08 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0140] The compound with a retention time of 4.22 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0141] The compound with a retention time of 1.42 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0142] The compound with a retention time of 2.71 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0143] The compound with a retention time of 1.11 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0144] The compound with a retention time of 2.21 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0145] The compound with a retention time of 3.88 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0146] The compound with a retention time of 4.42 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0147] The compound with a retention time of 1.254 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0148] The compound with a retention time of 2.267 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0149] The compound with a retention time of 4.473 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0150] The compound with a retention time of 4.688 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0151] The compound that has a retention time of 7.008 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0152] The compound with a retention time of 7.199 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0153] The compound with a retention time of 4.735 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0154] The compound with a retention time of 4.758 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0155] The compound with a retention time of 7.198 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0156] The compound with a retention time of 7.460 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0157] The compound with a retention time of 4.249 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0158] The compound with a retention time of 4.333 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0159] The compound with a retention time of 6.961 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0160] The compound with a retention time of 7.133 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0161] The compound with a retention time of 4.903 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0162] The compound with a retention time of 5.508 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0163] The compound with a retention time of 2.174 min under the following conditions is... One of the stereoisomers: Column: CHIRALART Cellulose SB, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0164] The compound with a retention time of 2.950 min under the following conditions is... One of the stereoisomers: Column: CHIRALART Cellulose SB, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0165] The compound with a retention time of 1.986 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IE, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0166] The compound with a retention time of 2.919 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IE, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0167] The compound with a retention time of 5.137 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0168] The compound with a retention time of 5.370 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40;

[0169] The compound with a retention time of 0.928 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0170] The compound with a retention time of 1.411 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0171] The compound with a retention time of 1.872 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE, Mobile phase B: MeOH:DCM=1:1;

[0172] The compound with a retention time of 2.467 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE, Mobile phase B: MeOH:DCM=1:1;

[0173] The compound with a retention time of 0.993 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1;

[0174] The compound with a retention time of 2.596 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE (0.5% 2mM NH3-MEOH), Mobile phase B: MeOH:DCM = 1:1.

[0175] The above retention time test conditions are not a limitation on the compound. As long as the above test conditions are used to determine the retention time, and the obtained retention time is the same as or within the error range described above, and the compound is a stereoisomer of the compound limited by the retention time described above, then it falls within the protection scope of this invention.

[0176] The present invention also provides a method for preparing the compound shown in Formula I, comprising the following steps:

[0177] The compounds shown in formula KX and K-XIII undergo substitution reactions to yield the compound shown in formula I.

[0178]

[0179] Among them, "*", n, ring A, and R 1 R 2 R 3 R 4 The definitions of X, Y, Z, U, V, W, and Q are as described in the previous item, and L is a halogen or hydroxyl group.

[0180] The operation and conditions of the substitution reaction can be conventional in the art, and preferably, carried out in the presence of a base (e.g., DIEA and / or TEA).

[0181] The method for preparing the compound as shown in Formula I may further include the following steps:

[0182] The compound shown in formula K-IX is deprotected to give the compound shown in formula KX;

[0183]

[0184] Among them, "*", n, ring A, and R 2 R 3 R 4The definitions of X, Y, Z, U, V, W, and Q are as described in the previous item, with PG being an amino protecting group.

[0185] In formula K-IX, the amino protecting group can be a common amino protecting group in the art, preferably p-tert-butoxycarbonyl (Boc) or benzyloxycarbonyl (Cbz).

[0186] The operation and conditions of the deprotection reaction can be conventional in the art, for example, carried out in the presence of an acid (e.g., trifluoroacetic acid and / or hydrochloric acid).

[0187] The present invention also provides a compound as shown in formula KX:

[0188]

[0189] Among them, "*", n, ring A, and R 2 R 3 R 4 The definitions of X, Y, Z, U, V, W, and Q are as described in the previous item.

[0190] Preferably, the compound represented by formula KX is any of the following compounds:

[0191]

[0192]

[0193]

[0194]

[0195]

[0196] The present invention also provides a pharmaceutical composition comprising the above-described compounds as shown in Formula I, their solvates, their prodrugs, their metabolites, or their pharmaceutically acceptable salts, and pharmaceutical excipients.

[0197] The present invention also provides the use of the compounds shown in Formula I, their solvates, their prodrugs, their metabolites, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions thereof, in the preparation of a medicament. The medicament is used to treat cancer.

[0198] Preferably, the cancer includes one or more of the following: lung cancer, colon cancer, pancreatic cancer, rectal cancer, lymphoma, esophageal cancer, ovarian cancer, glioma, cervical cancer, urothelial carcinoma, gastric cancer, endometrial cancer, liver cancer, bile duct cancer, breast cancer, leukemia, and melanoma.

[0199] The present invention also provides the compounds shown in Formula I, their solvates, their prodrugs, their metabolites, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions thereof in the preparation of KRAS. G12C Applications in protein mutation inhibitors.

[0200] In the aforementioned application, the KRAS G12C Protein mutation inhibitors can be used in mammalian organisms; they can also be used in vitro, primarily for experimental purposes, such as providing comparisons as standards or controls, or preparing kits according to conventional methods in the art, for KRAS. G12C The protein mutation inhibition effect can be rapidly detected.

[0201] Unless otherwise specified, the terms used in this invention have the following meanings:

[0202] Those skilled in the art will understand that, according to convention used in the art, the use of “” in the structural formula describing a group in this invention refers to the connection of the corresponding group to other segments or groups in the compound through that site. When “” is used in this invention to indicate a chemical bond connected to a single bond, it signifies a mixture of R and S configurations.

[0203] The term "comprising" is an open-ended expression, meaning it includes the contents specified in this invention, but does not exclude other aspects.

[0204] The term "multiple" refers to 2, 3, 4 or 5, preferably 2 or 3.

[0205] The term "room temperature" refers to 20–30°C.

[0206] The term "pharmaceutically acceptable" means that the salts, solvents, excipients, etc., are generally non-toxic, safe, and suitable for patient use. The term "patient" preferably refers to a mammal, and more preferably a human.

[0207] The term "solvate" refers to a substance formed by the combination of the compound of this invention with a stoichiometric or non-stoichiometric solvent. Solvent molecules in a solvate can exist in an ordered or disordered arrangement. The solvents include, but are not limited to, water, methanol, and ethanol.

[0208] The term "prodrug" refers to a derivative of the compound of the present invention that, when administered to a warm-blooded animal (e.g., a human), is converted into the compound of the present invention (the drug). Typical examples of prodrugs include compounds having biologically variable protecting groups on the functional portion of the active compound. Prodrugs include compounds that can be converted into the active compound through oxidation, reduction, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydration, alkylation, dealkylation, acylation, deacylation, phosphorylation, and dephosphorylation.

[0209] The term "metabolite" refers to the degradation product of the compounds of the present invention through one or more metabolic processes, which exert the desired biological activity.

[0210] The term "pharmaceutically acceptable salt" refers to a salt prepared from the compounds of this invention, their solvates, their prodrugs, or their metabolites with a relatively non-toxic, pharmaceutically acceptable acid or base. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting a sufficient amount of a pharmaceutically acceptable base with the neutral form of such compounds in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, lithium salts, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, zinc salts, bismuth salts, ammonium salts, and diethanolamine salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting a sufficient amount of a pharmaceutically acceptable acid with the neutral form of such compounds in a pure solution or a suitable inert solvent. The pharmaceutically acceptable acids include inorganic acids, including but not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, and sulfuric acid. The pharmaceutically acceptable acids include organic acids, including but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acidic citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentian acid, fumaric acid, gluconic acid, succinic acid, formic acid, ethanesulfonic acid, dihydroxynaphthyl acid (i.e., 4,4'-methylene-bis(3-hydroxy-2-naphthylcarboxylic acid)), amino acids (e.g., glutamic acid, arginine), etc. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. For details, see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977), or Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

[0211] The terms "compound," "solvate," "prodrug," "metabolite," and their "pharmaceutically acceptable salts" can exist in crystalline or amorphous forms. The term "crystalline form" refers to a state in which the ions or molecules are arranged in a strictly periodic manner in three-dimensional space, exhibiting a regular repetition at certain intervals; due to different periodsic arrangements, multiple crystalline forms can exist, a phenomenon known as polymorphism. The term "amorphous form" refers to a state in which the ions or molecules are distributed randomly, meaning that there is no periodic arrangement between the ions and molecules.

[0212] The terms "compound," "solvent," "prodrug," "metabolite," and their "pharmaceutically acceptable salts," if stereoisomers exist, may be present as a single stereoisomer or a mixture thereof (e.g., a racemic mixture). The term "stereoisomer" refers to cis-trans isomers or optical isomers. These stereoisomers can be separated, purified, and enriched by asymmetric synthetic methods or chiral separation methods (including but not limited to thin-layer chromatography, rotational chromatography, column chromatography, gas chromatography, high-performance liquid chromatography, etc.), and can also be obtained by chiral resolution through bonding (chemical bonding, etc.) or salt formation (physical bonding, etc.) with other chiral compounds. The term "single stereoisomer" means that the mass content of one stereoisomer of the compound of the present invention is not less than 95% relative to all stereoisomers of the compound.

[0213] The terms “compound,” “solvent,” “prodrug,” “metabolite,” and their “pharmaceuticalally acceptable salts” may exist as a single tautomer or a mixture thereof, preferably as the more stable tautomer.

[0214] The atoms in the terms "compound," "solvate," "prodrug," "metabolite," and their "pharmaceuticalally acceptable salts" can exist in either their natural abundance or non-natural abundance. For example, the natural abundance of a hydrogen atom means that approximately 99.985% of it is protium and approximately 0.015% is deuterium; its non-natural abundance means that approximately 95% of it is deuterium. That is, one or more atoms in the terms "compound," "solvate," "prodrug," "metabolite," and their "pharmaceuticalally acceptable salts" can be atoms existing in a non-natural abundance form.

[0215] When any variable (e.g., Re) appears multiple times in the definition of a compound, the definition of that variable at each position is independent of its definitions at other positions; their meanings are mutually independent and do not affect each other. Therefore, if a group is substituted by one, two, or three Re groups—that is, if the group may be substituted by up to three Re groups—the definition of Re at that position is independent of the definitions of Re at other positions. Furthermore, combinations of substituents and / or variables are only permitted if the combination produces a stable compound.

[0216] The term "alkyl" refers to a straight-chain or branched alkyl group having a specified number of carbon atoms.

[0217] The term "alkoxy" refers to the group -O-RX, where RX is an alkyl group as defined above.

[0218] The term "cycloalkyl" refers to a monovalent saturated cyclic alkyl group. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0219] The term "heterocyclic alkyl" refers to a saturated monocyclic or polycyclic group having a heteroatom. Examples of heterocyclic alkyl groups include: pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydropyridyl, tetrahydropyrrolidinyl, azaheptanyl, thiazolyl, azoleyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazine, azaheptanyl, diazaheptanyl, oxonitroheptanyl, etc.

[0220] The term "aryl" refers to C6-C10 aryl groups, such as phenyl or naphthyl.

[0221] The term "heteroaryl" refers to an aromatic group containing heteroatoms, preferably containing one, two, or three independent 5- or 6-membered monocyclic aromatic rings or 9- or 10-membered bicyclic aromatic rings selected independently from nitrogen, oxygen, and sulfur. When it is a bicyclic ring, at least one ring is aromatic. Examples of heteroaryl groups include: furanyl, pyridinyl, pyridinyl, pyrimidinyl, pyrazinyl, thiopheneyl, isozolyl, oxazolyl, diazolyl, imidazoleyl, pyrroleyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, benzimidazolyl, indolyl, inzolyl, benzothiazolyl, benziisothiazolyl, benzozolyl, benziisozolyl, quinolinyl, isoquinolinyl, etc.

[0222] In some cases, standard techniques are used in chemical synthesis, chemical analysis, and performance testing of light-emitting devices.

[0223] Furthermore, it should be noted that, unless otherwise explicitly stated, the descriptive phrase "...independently" used in this invention should be interpreted broadly, meaning that the described entities are independent of each other and can independently be the same or different specific functional groups. More specifically, the descriptive phrase "...independently" can mean either that the specific options expressed by the same symbol in different functional groups do not affect each other, or that the specific options expressed by the same symbol in the same functional group do not affect each other.

[0224] The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the dispensing of prescriptions. It includes all substances contained in pharmaceutical preparations, excluding the active ingredient. See also the Pharmacopoeia of the People's Republic of China (2015 Edition), Volume IV, or the Handbook of Pharmaceutical Excipients (Raymond C. Rowe, 2009 Sixth Edition).

[0225] The term “treatment” refers to a therapeutic approach. When a specific condition is involved, treatment means: (1) alleviating one or more biological manifestations of the disease or condition; (2) interfering with (a) one or more points in a biological cascade that causes or precipitates the condition or (b) one or more biological manifestations of the condition; (3) improving one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or its treatment; or (4) slowing the development of the condition or one or more biological manifestations of the condition.

[0226] The term "prevention" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0227] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain the preferred compounds of the present invention.

[0228] The reagents and raw materials used in this invention are all commercially available.

[0229] The positive and progressive effects of this invention are as follows: This invention provides a series of novel compounds with good KRAS properties. G12C It has protein mutation inhibitory activity and can be used to treat or prevent cancer.

[0230] The compounds of this invention exhibit strong cytotoxicity against a variety of tumor cells. For example, all the compounds of this invention exhibit proliferation inhibitory activity against NCI-H358, MIA PaCa-2 and other cells, with IC50 values ​​<100 μM. Attached Figure Description

[0231] Figure 1 X-Ray plot of intermediate A27. Detailed Implementation

[0232] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0233] In the following examples, the compounds were prepared using the following process:

[0234]

[0235] In the above compounds, L is a halogen (e.g., chlorine) or hydroxyl group, PG is a common amino protecting group (e.g., tert-butoxycarbonyl), and the definitions of the other substituents are as described in the preceding item.

[0236] The starting compound KI undergoes a substitution reaction to give K-II; K-II undergoes a halogenation reaction to give K-III; K-III and K-XI undergo a substitution reaction to give K-IV; K-IV undergoes deprotection to give KV; KV undergoes a halogenation reaction to give K-VI; K-VI and K-XII undergo a substitution reaction to give K-VII; K-VII undergoes a further substitution reaction to give K-VIII; K-VIII undergoes a reductive ammonolysis reaction (or a further substitution reaction) to give K-IX; K-IX undergoes deprotection to give KX; KX and K-XIII undergo a substitution or acylation reaction to give compound I; compound I undergoes chiral resolution by SFC to give compounds Ia and Ib. The operation and conditions of each step are all standard in the art.

[0237] In the following examples, the structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0238] SHIMADZU LC system (column: CSH TM Prep-C18, 19*150mm, Liquid Handling Unit LH-40, Pump LC-20AP, Detector SPD-20A, System Controller CBM-20A, Solvent System: Acetonitrile and 0.05% Trifluoroacetic Acid Aqueous Solution).

[0239] The LC / MS spectra of the compounds were obtained using LC / MS (Agilent Technologies 1200 Series). The LC / MS conditions were as follows (run time: 10 minutes):

[0240] Acidic conditions: A: 0.05% aqueous solution of trifluoroacetic acid; B: 0.05% acetonitrile solution of trifluoroacetic acid;

[0241] Alkaline conditions: A: 0.05% aqueous solution of NH3·H2O; B: acetonitrile

[0242] Neutral conditions: A: 10mM aqueous solution of NH4OAC; B: acetonitrile

[0243] Unless otherwise specified, in the following examples, intermediates and final compounds were purified using silica gel column chromatography or by using... CSH TM Prep-C18 (5μm, OBD) TM 19*150mm column or XBridge™ Prep Phenyl (5μm, OBD) TM The sample (30*100mm) was purified by preparative HPLC on a reversed-phase column.

[0244] Silica gel column chromatography generally uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0245] The CombiFlash rapid preparation system uses a CombiFlash Rf200 (TELEDYNE ISCO).

[0246] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The silica gel plates used for TLC detection products have a diameter of 0.15mm to 0.2mm, while those used for TLC separation and purification products have a diameter of 0.4mm to 0.5mm.

[0247] Unless otherwise specified, in the following examples, intermediates or final compounds were separated by SFC under the following conditions: Equipment: Waters SFC 150 preparative chromatograph; Column: CHIRAL ART Cellulose-SB, 5cm×25cm, 5μm or Dr. Maish Reprosil Chiral-JM, 250x25mm, 10μm; Column temperature: room temperature; Mobile phase: ethanol in n-hexane or CO2 / MeOH = 65 / 35; Flow rate: 70g / min; Wavelength: 214nm; Back pressure: 100bar.

[0248] The known starting materials of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0249] Abbreviations: AcOH: Acetic acid; ACN: Acetonitrile; AcCl: Acetyl chloride; Acrylamide: Acrylamide; AgF: Silver fluoride; Ag2O: Silver oxide; BH3: Borane; Boc2O: Tert-butoxycarbonyl tert-butyl carbonate; Boc: Tert-butoxycarbonyl; Conc.HCl: Concentrated hydrochloric acid; C2Br2Cl4: 1,2-Dibromotetrachloroethane; Cbz: Benzyloxycarbonyl; CH3I: Iodomethane; Cs2CO3: Cesium carbonate; DAST: Diethylaminotrifluoride Sulfur; DIEA: N,N-diisopropylethylamine; DMAP: 4-dimethylaminopyridine; DCM: dichloromethane; Dioxane: 1,4-dioxane; DMF: dimethylformamide; DMA: dimethylacetamide; DMS: dimethyl sulfoxide; DCE: 1,2-dichloroethane; DMF-DMA: 1,1-dimethoxy-N,N-dimethyl-methylamine; Dppf: bis(diphenylphosphine)ferrocene; EDCI: 1-ethyl-3(3-dimethylpropylamine)carbodiimide; Ethyl propiolate: ethyl propionate; EtOH: ethanol; Et3N: triethylamine; H2: hydrogen; HCOONH4: ammonium formate; LAH: lithium aluminum hydride; LiHMDS: lithium bis(trimethylsilyl)amino; LDA: lithium diisopropylamino; LiOH: lithium hydroxide; LiAlH4: lithium aluminum hydride; MeOH: methanol; MsC: methanesulfonyl chloride; MW: microwave reaction; m-CPBA: m-chloroperoxybenzoic acid; n-BuLi: n-butyllithium; NaH: sodium hydride; N aHMDS: Sodium bis(trimethylsilyl)amino; Ph3PCH2OMeCl: Methoxymethyltriphenylphosphine chloride; Pd2(dba)3: Tris(dibenzylacetone)dipalladium; Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; PPA: Polyphosphoric acid; POCl3: Phosphorus oxychloride; Pd / C: Palladium on carbon; Pyrrolidine: Pyrrolidine; SOCl2: Thionyl chloride; t-BuOK: Potassium tert-butoxide; t-butyl nitrite: tert-butyl nitrite; t-BuLi: tert-butyllithium; Toluene: toluene; TsOH: p-toluenesulfonic acid; THF: tetrahydrofuran; TFA: trifluoroacetic acid; TEA: triethylamine; Urea: urea; Xphos: 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl; Xantphos: 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene; Zn / Cu: zinc-copper reagent; Zn: zinc powder; Zn(CN)2: zinc cyanide.

[0250] Preparation of intermediate A1 in Example 1

[0251] 4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0252]

[0253] Step 1: Preparation of tert-butyl 4-(2-chloro-6-methyl-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid

[0254]

[0255] 2,4-Dichloro-6-methyl-5-nitropyrimidine (6 g, 28.8 mmol) was dissolved in anhydrous dichloromethane (30 mL), and piperazine-1-carboxylic acid tert-butyl ester (10.75 g, 57.7 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 4-(2-chloro-6-methyl-5-nitropyrimidine-4-yl)piperazine-1-carboxylic acid tert-butyl ester (6 g, 58% yield). MS: 358.1 (M+H) + . 1 H NMR (400MHz, Chloroform-d) δ3.54(s,8H),2.45(s,3H),1.49(s,9H).

[0256] Step 2: Preparation of tert-butyl 4-(2-chloro-6-methyl-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid

[0257]

[0258] 1.8 g (5.03 mmol) of tert-butyl 4-(2-chloro-6-methyl-5-nitropyrimidin-4-yl)piperazine-1-carboxylate was dissolved in anhydrous tetrahydrofuran (40 mL). LDA (6.04 mmol) was slowly added with stirring at -78 °C, and stirring continued for 30 min. Then, a solution of 1,2-dibromo-1,1,2,2-tetrachloroethane (1.966 g, 6.04 mmol) in tetrahydrofuran (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 4-(6-(bromomethyl)-2-chloro-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (1 g, 45% yield). MS: 436.1 & 438.1 (M+H) + .

[0259] Step 3: Preparation of tert-butyl 4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid

[0260]

[0261] Methyl 1,2,3,4-tetrahydronaphthalene-1-carboxylate (1.568 g, 8.24 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL). LDA (5.50 mmol) was slowly added with stirring at -78 °C, and stirring was continued at -78 °C for 30 min. Then, a solution of 4-(6-(bromomethyl)-2-chloro-5-nitropyrimidin-4-yl)piperazine-1-carboxylate tert-butyl (1.2 g, 2.75 mmol) in tetrahydrofuran (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (300 mg, 21% yield). MS: 546.1 (M+H) + .

[0262] Preparation of intermediate A2 in Example 2

[0263] (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester

[0264]

[0265] Step 1: Preparation of (S)-4-(2-chloro-6-methyl-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester

[0266]

[0267] 2,4-Dichloro-6-methyl-5-nitropyrimidine (5.0 g, 24.04 mmol) was dissolved in anhydrous dichloromethane (30 mL), and tert-butyl (S)-3-methylpiperazine-1-carboxylate (9.63 g, 48.1 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid (S)-4-(2-chloro-6-methyl-5-nitropyrimidine-4-yl)-3-methylpiperazine-1-carboxylate (8 g, 90% yield). MS: 372.1 (M+H) + .

[0268] Step 2: Preparation of (S)-4-(6-(bromomethyl)-2-chloro-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester

[0269]

[0270] (S)-4-(2-chloro-6-methyl-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (4 g, 10.76 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). LDA (12.91 mmol) was slowly added with stirring at -65 °C, and stirring continued for 40 min. Then, a solution of 1,2-dibromo-1,1,2,2-tetrachloroethane (4.20 g, 12.91 mmol) in tetrahydrofuran (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -65 °C for 0.5 h, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (S)-4-(6-(bromomethyl)-2-chloro-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (1.0 g, 21% yield). MS: 450.1 & 452.1 (M+H) + .

[0271] Step 3: Preparation of (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester

[0272]

[0273] Methyl 1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (169 mg, 0.887 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL). LDA (0.887 mmol) was slowly added with stirring at -78 °C, and stirring was continued at -78 °C for 30 min. Then, a solution of (S)-4-(6-(6-bromomethyl)-2-chloro-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.444 mmol) in tetrahydrofuran (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (140 mg, 56.3% yield). MS: 561.1 (M+H) + .

[0274] Preparation of intermediate A3 in Example 3

[0275] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0276]

[0277] Step 1: Preparation of 2,4-bis(benzyloxy)-6-methyl-5-nitropyrimidine

[0278]

[0279] 2,4-Dichloro-6-methyl-5-nitropyrimidine (20 g, 96 mmol) and phenylmethanol (52.0 g, 481 mmol) were dissolved in anhydrous tetrahydrofuran (40 mL), and potassium tert-butoxide (21.58 g, 192 mmol) was added with stirring under reflux. After the addition was complete, the reaction mixture was stirred for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give 2,4-bis(benzyloxy)-6-methyl-5-nitropyrimidine (16 g, 47.4% yield) as a white solid. MS: 352.1 (M+H) + .

[0280] Step 2: Preparation of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine

[0281]

[0282] 15 g (42.7 mmol) of 2,4-bis(benzyloxy)-6-methyl-5-nitropyrimidine was dissolved in anhydrous tetrahydrofuran (500 mL). LiHMDS (68.3 mmol) was slowly added with stirring at -78 °C for 40 min. Then, a solution of 13.90 g (42.7 mmol) in tetrahydrofuran (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine (12.8 g, 70% yield). MS:430.1&432.1(M+H) + .

[0283] Step 3: Preparation of methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0284]

[0285] Methyl 1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (6.63 g, 34.9 mmol) was dissolved in anhydrous tetrahydrofuran (150 mL). LDA (34.9 mmol) was slowly added with stirring at -78 °C, and stirring was continued at -78 °C for 30 min. Then, a solution of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine (5 g, 11.62 mmol) in tetrahydrofuran (15 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a white solid, methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (4.5 g, 72% yield). MS: 540.1 (M+H) + .

[0286] Step 4: Preparation of methyl 1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0287]

[0288] Methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.0 g, 1.853 mmol) was dissolved in trifluoroacetic acid (5 mL). The reaction mixture was slowly heated to 100 °C under nitrogen protection and stirred in a microwave oven for 1 h. The reaction mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid methyl 1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (600 mg, 90% yield). MS: 360.1 (M+H) + .

[0289] Step 5: Preparation of methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0290]

[0291] Methyl 1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (2.2 g, 6.12 mmol) was dissolved in phosphorus oxychloride (15 mL), and DIEA (791 mg, 6.12 mmol) was added with stirring at 0°C. The reaction mixture was slowly heated to 110°C under nitrogen protection and stirred for 6 hours. The reaction mixture was then concentrated, diluted with dichloromethane for extraction, and the solution was poured into a saturated sodium bicarbonate solution and stirred for 1 hour. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The solution was then filtered and concentrated to give methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (2.2 g, 91% yield) as a yellow solid. MS: 397.2 (M+H) + .

[0292] Step 6: Preparation of (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0293]

[0294] 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (1.5 g, 3.79 mmol) was dissolved in anhydrous dichloromethane (30 mL), and (S)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (1.71 g, 7.57 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (1.0 g, 45% yield). MS: 586.1 (M+H) + .

[0295] Preparation of intermediate A4 in Example 4

[0296] (2R,6S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

[0297]

[0298] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 0.5 g, 1.3 mmol) was dissolved in anhydrous dichloromethane (30 mL), and (2R,6S)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (0.57 g, 2.6 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2R,6S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (0.402 g, 54% yield). MS: 574.1 (M+H) + .

[0299] Preparation of intermediate A5 in Example 5

[0300] (2R,5S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylic acid tert-butyl ester

[0301]

[0302] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 0.5 g, 1.3 mmol) was dissolved in anhydrous dichloromethane (30 mL), and tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (0.57 g, 2.6 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2R,5S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylic acid tert-butyl ester (0.342 g, 46% yield). MS: 574.1 (M+H) + .

[0303] Preparation of intermediate A6 in Example 6

[0304] 7-(2-chloro-6-(((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester

[0305]

[0306] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 0.5 g, 1.3 mmol) was dissolved in anhydrous dichloromethane (30 mL), and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylic acid (0.588 g, 2.6 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 7-(2-chloro-6-(((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (0.388 g, 51% yield). MS: 587.1 (M+H) + .

[0307] Preparation of intermediate A7 in Example 7

[0308] (1R, 5S)-3-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0309]

[0310] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 0.5 g, 1.3 mmol) was dissolved in anhydrous dichloromethane (30 mL), and (1R,5S)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (0.551 g, 2.6 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (1R,5S)-3-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (0.56 g, 75.1% yield). MS: 573.1 (M+H) + .

[0311] Preparation of intermediate A8 in Example 8

[0312] (1R, 5S)-8-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester

[0313]

[0314] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 0.5 g, 1.3 mmol) was dissolved in anhydrous dichloromethane (30 mL). (1R,5S)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester (0.551 g, 2.6 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (1R,5S)-8-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester (0.49 g, 65.9% yield). MS: 573.1 (M+H) + .

[0315] Preparation of intermediate A9 in Example 9

[0316] (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester

[0317]

[0318] Step 1: Preparation of (R)-3-(fluoromethyl)piperazine-1-carboxylic acid tert-butyl ester

[0319]

[0320] (R)-3-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (2 g, 9.25 mmol) was dissolved in anhydrous dichloromethane (30 mL). While stirring at 0 °C, a solution of N-ethyl-N-(trifluorosulfonyl)ethylamine (1.79 g, 11.10 mmol, 1.47 mL) in dichloromethane (2 mL) was slowly added. After the addition was complete, the reaction mixture was slowly brought to room temperature and stirred for 3 hours. Once the reaction was complete, the reaction solution was used directly for the next reaction step.

[0321] Step 2: Preparation of 4-(tert-butyl)(R)-2-(fluoromethyl)piperazine-1-benzyl-1,4-dicarboxylic acid ester

[0322]

[0323] Under stirring at 0 °C, N-ethyl-N-isopropyl-propane-2-amine (1.19 g, 9.21 mmol, 1.60 mL) and benzyl 2,5-dioxopyrrolidine-1-carboxylate (3.22 g, 13.81 mmol) were slowly added to a solution of (R)-3-(fluoromethyl)piperazine-1-carboxylate tert-butyl ester (9.25 mmol) in dichloromethane (30 mL). After addition, the reaction mixture was slowly raised to room temperature and stirred for 3 hours. The reaction was then quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid 4-(tert-butyl)(R)-2-(fluoromethyl)piperazine-1-benzyl-1,4-dicarboxylate (1.02 g, 31.4% yield). MS: 353.1 (M+H) + . 1 H NMR (400MHz, CDCl3) δ7.41–7.29(m,5H),5.21–5.10(m,2H),4.55–4.35(m,3H),4.19–3.82(m,3H),3.16–2.98(m,3H),2.89(s,1H),1.47(s,9H).

[0324] Step 3: Preparation of (R)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester

[0325]

[0326] 4-(tert-butyl)(R)-2-(fluoromethyl)piperazine-1-benzyl-1,4-dicarboxylic acid ester (300 mg, 0.851 mmol) was dissolved in a mixed solution of dichloromethane (5 mL) and TFA (2 mL). The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was diluted with ethyl acetate for extraction. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance (R)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester (190 mg, 88% yield). MS: 253.1 (M+H) + .

[0327] Step 4: (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester

[0328]

[0329] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 0.3 g, 0.757 mmol) was dissolved in anhydrous dichloromethane (10 mL), and (R)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester (191 mg, 0.757 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester (185 mg, 39.9% yield). MS: 613.1 (M+H) + .

[0330] Preparation of intermediate A10 in Example 10

[0331] (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0332]

[0333]

[0334] Step 1: Preparation of 1-benzyl 4-(tert-butyl)(R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylic acid ester

[0335]

[0336] (3R)-3-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (5.2 g, 24.04 mmol) was dissolved in a mixture of dichloromethane (20 mL) and water (20 mL). NaHCO3 (6.1 g, 72.62 mmol) was added with stirring at 0 °C, followed by the slow addition of a dichloromethane (10 mL) solution of benzyl chloroformate (4.3 g, 25.21 mmol). After the addition was complete, the reaction mixture was brought to room temperature and stirred for 18 hours. The reaction was then quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 1-benzyl-4-(tert-butyl)(R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylic acid ester (9.1 g, 108.0% yield). MS:351.1&251.1(M+H) + .

[0337] Step 2: Preparation of 1-benzyl 4-(tert-butyl)(R)-2-(((methanesulfonyl)oxy)methyl)piperazine-1,4-dicarboxylic acid ester

[0338]

[0339] 1-Benzyl 4-(tert-butyl)(R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylic acid ester (5.3 g, 15.13 mmol) and DIEA (3.91 g, 30.25 mmol, 5.27 mL) were dissolved in anhydrous dichloromethane (30 mL). A solution of MsCl (2.60 g, 22.69 mmol) in dichloromethane (10 mL) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, 1-benzyl 4-(tert-butyl)(R)-2-(((methanesulfonyl)oxy)methyl)piperazine-1,4-dicarboxylic acid ester (5.3 g, 81.78% yield). MS:429.1&329.1(M+H) + .

[0340] Step 3: Preparation of 1-benzyl 4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylic acid ester

[0341]

[0342] 1-Benzyl 4-(tert-butyl)(R)-2-(((methanesulfonyl)oxy)methyl)piperazine-1,4-dicarboxylic acid ester (5.3 g, 12.37 mmol) and potassium cyanide (KCN) (1.61 g, 24.72 mmol) were dissolved in DMA (40 mL). The reaction mixture was slowly heated to 80 °C and stirred for 18 hours under nitrogen protection, then diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil, 1-benzyl 4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylic acid ester (4 g, 89.9% yield). MS: 360.1 & 304.1 (M+H) + .

[0343] Step 4: Preparation of (S)-3-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0344]

[0345] 2.4 g (6.68 mmol) of 1-benzyl-4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylic acid ester was dissolved in ethanol (5 mL). Palladium on carbon (711 mg) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature for 1 hour under hydrogen atmosphere, and then filtered. The filtrate was concentrated under vacuum to give a yellow solid (S)-3-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (1.5 g, 100% yield). MS: 226.1 (M+H) + .

[0346] Step 5: Preparation of (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0347]

[0348] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 600 mg, 1.514 mmol) was dissolved in anhydrous dichloromethane (10 mL), and tert-butyl 1-(S)-3-(cyanomethyl)piperazine-1-carboxylic acid (341 mg, 1.514 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (450 mg, 50.8% yield). MS: 586.1 (M+H) + .

[0349] Preparation of intermediate A11 in Example 11

[0350] (2S)-2-(cyanomethyl)-4-(2-methoxy-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0351]

[0352] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 150 mg, 0.256 mmol) was dissolved in anhydrous methanol (5 mL), and sodium methoxide (27.7 mg, 0.513 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was brought to room temperature and stirred for 1 hour. The reaction was then quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(2-methoxy-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (0.12 g, 100% yield). MS: 581.1 (M+H) + .

[0353] Preparation of intermediate A12 in Example 12

[0354] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0355]

[0356]

[0357] Step 1: Preparation of methyl 2,3-dihydro-1H-indene-1-carboxylate

[0358]

[0359] 11 g (67.8 mmol) of 2,3-dihydro-1H-indene-1-carboxylic acid was dissolved in 100 mL of anhydrous methanol. Thionyl chloride (40.3 g, 339 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly heated to 70 °C and stirred for 1 hour. After the reaction was complete, the reaction mixture was concentrated and extracted with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography to give methyl 2,3-dihydro-1H-indene-1-carboxylic acid (11 g, 92.1% yield) as a colorless oil. MS: 177.1 (M+H) + .

[0360] Step 2: Preparation of methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid

[0361]

[0362] Methyl 2,3-dihydro-1H-indene-1-carboxylic acid (1.3 g, 6.97 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). LDA (6.97 mmol) was slowly added with stirring at -78 °C, and stirring was continued at -78 °C for 30 min. Then, a tetrahydrofuran solution of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine (intermediate A3-2, 1.0 g, 2.32 mmol) (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid (670 mg, 55% yield). MS: 526.1 (M+H)+ .

[0363] Step 3: Preparation of methyl 1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid

[0364]

[0365] Methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid (2.0 g, 3.81 mmol) was dissolved in trifluoroacetic acid (5 mL). The reaction mixture was slowly heated to 100 °C under nitrogen protection and stirred in a microwave oven for 1 h. The reaction mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid methyl 1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid (1.3 g, 99% yield). MS: 346.1 (M+H) + .

[0366] Step 4: Preparation of methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid

[0367]

[0368] Methyl 1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid (600 mg, 1.738 mmol) was dissolved in phosphorus oxychloride (15 mL), and DIEA (225 mg, 1.73 mmol) was added with stirring at 0°C. The reaction mixture was slowly heated to 110°C under nitrogen protection and stirred for 6 hours. The reaction mixture was then concentrated, diluted with dichloromethane for extraction, and the solution was poured into a saturated sodium bicarbonate solution and stirred for 1 hour. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The solution was then filtered and concentrated to give methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid (600 mg, 90% yield) as a yellow solid. MS: 383.1 (M+H) + .

[0369] Step 5: Preparation of (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0370]

[0371] 1-Benzyl-4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylic acid ester (intermediate A10 / step 3, 10.0 g, 27.85 mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (15 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to dryness and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution, saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a yellow solid (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (4.2 g, 58% yield). MS: 260.1 (M+H) + .

[0372] Step 6: Preparation of (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0373]

[0374] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-2,3-dihydro-1H-indene-1-carboxylic acid (600 mg, 1.570 mmol) was dissolved in anhydrous dichloromethane (10 mL), and (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (814 mg, 3.14 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (300 mg, 31.6% yield). MS: 606.1 (M+H) + .

[0375] Preparation of intermediate A13 in Example 13

[0376] Benzyl(2S)-4-(2-chloro-6-((4-(methoxycarbonyl)chroman-4-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0377]

[0378] Step 1: Preparation of methyl benzodihydropyran-4-carboxylate

[0379]

[0380] Benzodihydropyran-4-carboxylic acid (1 g, 5.61 mmol) was dissolved in anhydrous methanol (10 mL), and thionyl chloride (1.335 g, 11.22 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly heated to 70 °C and stirred for 1 hour. After the reaction was complete, the reaction mixture was concentrated and extracted with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography to give a colorless oil, methyl benzodihydropyran-4-carboxylate (0.6 g, 55.6% yield). MS: 193.1 (M+H) + .

[0381] Step 2: Preparation of methyl 4-((2,6-di(benzyloxy)-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid

[0382]

[0383] Methyl benzodihydropyran-4-carboxylate (300 mg, 1.561 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL). LDA (1.87 mmol) was slowly added with stirring at -78 °C, and stirring was continued at -78 °C for 30 min. Then, a tetrahydrofuran solution of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine (intermediate A3-2,672 mg, 1.561 mmol) (10 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78 °C for 0.5 h, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product, which was purified by silica gel column chromatography to give a yellow solid, methyl 4-((2,6-di(benzyloxy)-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid (420 mg, 50% yield). MS: 542.1 (M+H) + .

[0384] Step 3: Preparation of methyl 4-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid

[0385]

[0386] Methyl 4-((2,6-di(benzyloxy)-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid (300 mg, 0.554 mmol) was dissolved in trifluoroacetic acid (5 mL). The reaction mixture was slowly heated to 100 °C under nitrogen protection and stirred in a microwave oven for 1 hour. The reaction mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid methyl 4-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid (194 mg, 97% yield). MS: 362.1 (M+H) + .

[0387] Step 4: Preparation of methyl 4-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid

[0388]

[0389] Methyl 4-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid (280 mg, 0.775 mmol) was dissolved in phosphorus oxychloride (5 mL), and DIEA (100 mg, 0.775 mmol) was added with stirring at 0°C. The reaction mixture was slowly heated to 110°C under nitrogen protection and stirred for 6 hours. The reaction mixture was then concentrated, diluted with dichloromethane for extraction, and the solution was poured into a saturated sodium bicarbonate solution and stirred for 1 hour. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The solution was then filtered and concentrated to give methyl 4-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid (309 mg, 100% yield) as a yellow solid. MS: 399.1 (M+H) + .

[0390] Step 5: Preparation of benzyl(2S)-4-(2-chloro-6-((4-(methoxycarbonyl)chroman-4-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0391]

[0392] 4-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)benzodihydropyran-4-carboxylic acid methyl ester (309 mg, 0.776 mmol) was dissolved in anhydrous dichloromethane (10 mL), and (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A12 / step 5, 402 mg, 1.55 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, benzyl(2S)-4-(2-chloro-6-((4-(methoxycarbonyl)chroman-4-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (120 mg, 24.9% yield). MS: 622.1 (M+H) + .

[0393] Preparation of intermediate A14 in Example 14

[0394] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0395]

[0396]

[0397] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 1.5 g, 3.79 mmol) was dissolved in anhydrous dichloromethane (30 mL). Benzyl (S)-2-(cyanomethyl)piperazine-1-carboxylic acid (intermediate A12 / step 5, 1.96 g, 7.57 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (1.35 g, 58% yield). MS: 620.1 (M+H) + .

[0398] Preparation of intermediate A15 in Example 15

[0399] (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester

[0400]

[0401] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 200 mg, 0.51 mmol) was dissolved in anhydrous dichloromethane (5 mL), and (R)-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (164 mg, 0.76 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a colorless powder (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (180 mg, 62% yield). MS: 577.1 (M+H) + .

[0402] Preparation of intermediate A16 in Example 16

[0403] 6-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,6-diazaspiro[3,4]octane-2-carboxylic acid tert-butyl ester

[0404]

[0405] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 200 mg, 0.51 mmol) was dissolved in anhydrous dichloromethane (5 mL), and tert-butyl 2,6-diazaspiro[3,4]octane-2-carboxylic acid (107 mg, 0.505 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, 6-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,6-diazaspiro[3,4]octane-2-carboxylic acid tert-butyl ester (145 mg, 51.9% yield). MS: 573.1 (M+H) + .

[0406] Preparation of intermediate A17 in Example 17

[0407] 7-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester

[0408]

[0409]

[0410] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 200 mg, 0.51 mmol) was dissolved in anhydrous dichloromethane (5 mL), and tert-butyl 2,7-diazaspiro[4,4]nonane-2-carboxylic acid (114 mg, 0.505 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow oily substance, 7-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (145 mg, 49% yield). MS: 587.1 (M+H) + .

[0411] Preparation of intermediate A18 in Example 18

[0412] (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitro-2-(pyridin-3-yl)pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester

[0413]

[0414] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14, 600 mg, 0.969 mmol) was dissolved in 1,4-dioxane (10 mL) and water (1 mL). Pyridine-3-boronic acid benzoyl ester (397 mg, 1.93 mmol), Pd(Ph3P)4 (112 mg, 0.097 mmol), and K2CO3 (402 mg, 2.91 mmol) were slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 80 °C and stirred for 3 hours. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitro-2-(pyridin-3-yl)pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (500 mg, 78% yield). MS: 662.1 (M+H) + .

[0415] Preparation of intermediate A19 in Example 19

[0416] (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(2-methylpyridin-3-yl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0417]

[0418] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14, 100 mg, 0.162 mmol) was dissolved in 1,4-dioxane (10 mL) and water (1 mL). 2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)pyridine (71 mg, 0.323 mmol), Pd(Ph3P)4 (18.7 mg, 0.016 mmol), and K2CO3 (67 mg, 0.485 mmol) were slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly heated to 80 °C and stirred for 3 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(2-methylpyridin-3-yl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (100 mg, 92% yield). MS: 676.1 (M+H) + .

[0419] Preparation of intermediate B1 in Example 20

[0420] 2-Fluoroacryl chloride

[0421]

[0422] 2-Fluoroacrylate (50 mg, 0.555 mmol) was dissolved in anhydrous dichloromethane (3 mL), and oxalyl chloride (106 mg, 0.833 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour. Once the reaction was complete, the reaction solution was used directly for the next reaction step.

[0423] Preparation of intermediate B2 in Example 21

[0424] Butyl-2-acetylacetyl chloride

[0425]

[0426] Butyric acid (300 mg, 3.57 mmol) was dissolved in anhydrous dichloromethane (3 mL), and oxaloyl chloride (679 mg, 5.35 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour. After the reaction was complete, the reaction solution was used directly for the next reaction.

[0427] Preparation of intermediate B3 in Example 22

[0428] (E)-4,4,4-trifluorobut-2-enoyl chloride

[0429]

[0430] (E)-4,4,4-trifluorobut-2-enoic acid (200 mg, 1.428 mmol) was dissolved in anhydrous dichloromethane (3 mL), and oxalyl chloride (544 mg, 4.28 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour. Once the reaction was complete, the reaction solution was used directly for the next reaction step.

[0431] Preparation of intermediate B4 in Example 23

[0432] (E)-4-methoxybut-2-enoyl chloride

[0433]

[0434] (E)-4-methoxybut-2-enoic acid (200 mg, 1.722 mmol) was dissolved in anhydrous dichloromethane (3 mL), and oxalyl chloride (219 mg, 1.722 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour. Once the reaction was complete, the reaction solution was used directly for the next reaction step.

[0435] Preparation of intermediate B5 in Example 24

[0436] (E)-4-Fluorobut-2-enoyl chloride

[0437]

[0438] Step 1: Preparation of (E)-4-fluorobut-2-enoic acid methyl ester

[0439]

[0440] Silver fluoride (4.25 g, 33.5 mmol) was dissolved in acetonitrile (20 mL), and a solution of (E)-4-bromobut-2-enoate methyl ester (2 g, 11.17 mmol) in acetonitrile (50 mL) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C in the dark for 48 hours. The solvent was then evaporated to dryness, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated sodium bicarbonate solution, saturated brine, dried anhydrous, and dried over sodium sulfate. The mixture was then filtered and concentrated to give (E)-4-fluorobut-2-enoate methyl ester (1.2 g, 91% yield).

[0441] Step 2: Preparation of (E)-4-fluorobut-2-enoic acid

[0442]

[0443] Methyl (E)-4-fluorobut-2-enoic acid (1.2 g, 10.16 mmol) was dissolved in tetrahydrofuran (30 mL), and a solution of lithium hydroxide (1.217 g, 50.8 mmol) in water (30 mL) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 4 hours, then the solvent was evaporated to dryness, and the mixture was extracted with dichloromethane. The organic phase was washed with dilute hydrochloric acid solution, saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow solid (E)-4-fluorobut-2-enoic acid (560 mg, 53% yield).

[0444] Step 3: Preparation of (E)-4-fluorobut-2-enoyl chloride

[0445]

[0446] (E)-4-fluorobut-2-enoic acid (208 mg, 2 mmol) was dissolved in anhydrous dichloromethane (3 mL), and oxalyl chloride (219 mg, 1.722 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour. Once the reaction was complete, the reaction solution was used directly for the next reaction step.

[0447] Preparation of intermediate C1 in Example 25

[0448] ((2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methanol

[0449]

[0450] 1-(tert-butyl)-2-methyl(2S,4S)-4-fluoropyrrolidine-1,2-dicarboxylic acid ester (10 g, 40.4 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), and LAH solution (162 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly raised to room temperature and stirred for 16 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil ((2S,4S)-4-fluoro-1-methylpyrrolidine-2-yl)methanol (2.2 g, 40% yield). MS: 134.1 (M+H) + .

[0451] Preparation of intermediate C2 in Example 26

[0452] ((2S,4R)-4-methoxy-1-methylpyrrolidone-2-yl)methanol

[0453]

[0454] Step 1: Preparation of 1-(tert-butyl)-2-methyl(2S,4R)-4-methoxypyrrolidine-1,2-dicarboxylic acid ester

[0455]

[0456] 1-(tert-butyl)-2-methyl(2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylic acid ester (5 g, 20.39 mmol) was dissolved in anhydrous acetonitrile (130 mL). Silver oxide (14.17 g, 61.2 mmol) and methyl iodoform (28.9 g, 204 mmol) were slowly added with stirring at 0 °C. After addition was complete, the reaction mixture was slowly brought to room temperature and stirred for 24 hours. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 1-(tert-butyl)-2-methyl(2S,4R)-4-methoxypyrrolidine-1,2-dicarboxylic acid ester (3.1 g, 59.6% yield). MS: 260.1 (M+H) + .

[0457] Step 2: Preparation of ((2S,4R)-4-methoxy-1-methylpyrrolidine-2-yl)methanol

[0458]

[0459] 3.2 g (12.34 mmol) of 1-(tert-butyl)-2-methyl(2S,4R)-4-methoxypyrrolidine-1,2-dicarboxylic acid ester was dissolved in anhydrous tetrahydrofuran (80 mL), and 62 mmol of LAH solution was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly raised to room temperature and stirred for 16 hours. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil (620 mg, 24.6% yield). MS: 146.1 (M+H) + .

[0460] Preparation of intermediate C3 in Example 27

[0461] ((2S,4R)-4-fluoro-1-methylpyrrolidone-2-yl)methanol

[0462]

[0463]

[0464] 1-(tert-butyl)-2-methyl(2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylic acid ester (10 g, 40.4 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), and LAH solution (162 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly raised to room temperature and stirred for 16 hours. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil ((2S,4R)-4-fluoro-1-methylpyrrolidine-2-yl)methanol (4.3 g, 80% yield). MS: 134.1 (M+H) + .

[0465] Preparation of intermediate C4 in Example 28

[0466] (3R,4R)-4-methoxy-1-methylpyrrolidine-3-ol

[0467]

[0468] (3R,4R)-3-hydroxy-4-methoxypyrrolidine-1-carboxylic acid tert-butyl ester (500 mg, 2.301 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), and LAH solution (4.6 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly raised to room temperature and stirred for 16 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil (3R,4R)-4-methoxy-1-methylpyrrolidine-3-ol (260 mg, 86% yield). MS: 132.1 (M+H) + .

[0469] Preparation of intermediate C5 in Example 29

[0470] (S)-(1-Methylpyrrolid-2-yl)methanethiol

[0471]

[0472] Step 1: Preparation of (S)-(1-methylpyrrolidone-2-yl)methylsulfonate

[0473]

[0474] (S)-(1-methylpyrrolidone-2-yl)methanol (2 g, 17.36 mmol) and TEA (5.27 g, 52.1 mmol) were dissolved in anhydrous dichloromethane (100 mL). Methanesulfonyl chloride (2.387 g, 20.84 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 3 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oily substance (S)-(1-methylpyrrolidone-2-yl)methanesulfonate (3.1 g, 92% yield). MS: 194.1 (M+H) + .

[0475] Step 2: Preparation of (S)-S-((1-methylpyrrolidone-2-yl)methyl)ethanethiol ester

[0476]

[0477] Methyl (S)-(1-methylpyrrolidone-2-yl)methanesulfonate (3.2 g, 16.56 mmol) was dissolved in anhydrous DMF (10 mL), and potassium thioacetate (2.08 g, 18.21 mmol) was added at 20 °C. After the addition was complete, the reaction mixture was stirred at 65 °C for 13 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give an orange oil (S)-S-((1-methylpyrrolidone-2-yl)methyl)ethanethiol ester (2.6 g, 91% yield). MS: 174.1 (M+H) + .

[0478] Step 3: Preparation of (S)-(1-methylpyrrolidone-2-yl)methanethiol

[0479]

[0480] (S)-S-((1-methylpyrrolidone-2-yl)methyl)ethanethiol ester (2.6 g, 15.01 mmol) was dissolved in methanol (40 mL), and sodium hydroxide (1.8 g, 45 mmol) was added at 20 °C. After the addition was complete, the reaction mixture was stirred at 25 °C for 13 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate, filtered, and concentrated to give a yellow oil (S)-(1-methylpyrrolidone-2-yl)methanethiol (620 mg, 31.5% yield). MS: 132.1 (M+H) + .

[0481] Preparation of intermediate A20 in Example 30

[0482] (2S,5R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)-5-methylpiperazine-1-carboxylic acid benzyl ester

[0483]

[0484]

[0485] Step 1: Preparation of ((benzyloxy)carbonyl)-L-serine-L-alanine methyl ester

[0486]

[0487] D-alanine methyl ester (25 g, 179.11 mmol) and ((benzyloxy)carbonyl)-L-serine (42.8 g, 179.11 mmol) were dissolved in dichloromethane (780 mL). EDCI (41.2 g, 214.93 mmol) and DIEA (109 mL, 626.89 mmol) were slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 10 °C for 12 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a white solid ((benzyloxy)carbonyl)-L-serine-L-alanine methyl ester (44.9 g, 77% yield). MS: 325.1 (M+H) + .

[0488] Step 2: Preparation of (3S,6R)-3-(hydroxymethyl)-6-methylpiperazine-2,5-dione

[0489]

[0490] Methyl ((benzyloxy)carbonyl)-L-serine-L-alanine (22.4 g, 69.07 mmol) was dissolved in methanol (70 mL) and cyclohexene (45 mL). Palladium on carbon (1.15 g) was added to the reaction mixture under stirring. The reaction mixture was stirred at 70 °C for 12 h under hydrogen atmosphere, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a gray solid (3S,6R)-3-(hydroxymethyl)-6-methylpiperazine-2,5-dione (7.9 g, 72.3% yield). 1H NMR (400MHz, DMSO) δ8.09 (s, 1H), 7.91 (s, 1H), 5.12 (t, J = 5.2Hz, 1H), 3.92–3.87 (m ,1H),3.74–3.70(m,1H),3.68–3.62(m,1H),3.52–3.48(m,1H),1.23–1.19(m,3H).

[0491] Step 3: Preparation of ((2R,5R)-5-methylpiperazin-2-yl)methanol

[0492]

[0493] (3S,6R)-3-(hydroxymethyl)-6-methylpiperazin-2,5-dione (7.9 g, 50 mmol) was dissolved in anhydrous tetrahydrofuran (150 mL), and a tetrahydrofuran solution of borane (1 M, 375 mL) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was slowly raised to 70 °C and stirred for 16 hours. The reaction was then quenched with methanol, followed by the addition of 5 M hydrochloric acid solution (27 mL). The reaction mixture was then slowly raised to 70 °C and stirred for 2 hours. Upon cooling, a solid precipitated, which was then filtered and dried to give a gray solid ((2R,5R)-5-methylpiperazin-2-yl)methanol (5.2 g, 80.8% yield). 1 H NMR (400MHz, DMSO) δ9.90(s,2H),5.61(s,1H),3.75–3.40(m,6H),3.18–3.02(m,2H),1.31–1.28(m,3H).

[0494] Step 4: Preparation of (2R,5R)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

[0495]

[0496] ((2R,5R)-5-methylpiperazin-2-yl)methanol (6.1 g, 30.03 mmol) was dissolved in methanol (50 mL). Diethylamine (9.43 g, 93.21 mmol, 13 mL) and Boc₂O (13.8 g, 63.23 mmol) were slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 50 °C for 12 hours. The reaction mixture was then concentrated to dryness, and the concentrate was dissolved in ethanol (100 mL). 1.5 M NaOH aqueous solution (100 mL) was added to the above solution, and the mixture was stirred at 100 °C for 12 hours. The reaction mixture was cooled and the pH was adjusted to 9 with 2 M hydrochloric acid solution, followed by extraction with dichloromethane. The organic phase was washed with a saturated aqueous solution of sodium bicarbonate and saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow solid (2R,5R)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (6 g, 86.5% yield). MS: 231.1 (M+H) + .

[0497] Step 5: 1-Benzyl 4-(tert-butyl)(2R,5R)-2-(hydroxymethyl)-5-methylpiperazine-1,4-dicarboxylic acid ester

[0498]

[0499] (2R,5R)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (7.1 g, 30.8 mmol) was dissolved in dichloromethane (80 mL). DIEA (4.38 g, 33.91 mmol, 5.91 mL) and CbzCl (5.52 g, 32.37 mmol) were slowly added with stirring at 0 °C. After addition was complete, the reaction mixture was stirred at 10 °C for 12 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a colorless oil, 1-benzyl-4-(tert-butyl)(2R,5R)-2-(hydroxymethyl)-5-methylpiperazine-1,4-dicarboxylic acid ester (8.9 g, 79.2% yield). MS: 265.1 (M+H) + .

[0500] Step 6: Preparation of 1-benzyl 4-(tert-butyl)(2R,5R)-5-methyl-2-((methanesulfonyl)oxy)methyl)piperazine-1,4-dicarboxylic acid ester

[0501]

[0502] 1-Benzyl 4-(tert-butyl)(2R,5R)-2-(hydroxymethyl)-5-methylpiperazine-1,4-dicarboxylic acid ester (8.9 g, 24.42 mmol) was dissolved in dichloromethane (30 mL). DIEA (6.31 g, 48.84 mmol, 8.51 mL) and MsCl (4.20 g, 36.63 mmol) were slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 2 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a yellow oil, 1-benzyl 4-(tert-butyl)(2R,5R)-5-methyl-2-((methanesulfonyl)oxy)methyl)piperazine-1,4-dicarboxylic acid ester (10.8 g, 100% yield).

[0503] Step 7: Preparation of 1-benzyl 4-(tert-butyl)(2S,5R)-2-(cyanomethyl)-5-methylpiperazine-1,4-dicarboxylic acid ester

[0504]

[0505] 1-Benzyl-4-(tert-butyl)(2R,5R)-5-methyl-2-((methanesulfonyl)oxy)methyl)piperazine-1,4-dicarboxylic acid ester (10.8 g, 24.41 mmol) was dissolved in DMA (100 mL), and potassium cyanide (4.77 g, 73.22 mmol) was slowly added with stirring at 20 °C. After the addition was complete, the reaction mixture was stirred at 80 °C for 12 hours, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil, 1-benzyl-4-(tert-butyl)(2S,5R)-2-(cyanomethyl)-5-methylpiperazine-1,4-dicarboxylic acid ester (5.0 g, 54.8% yield). MS: 374.1 & 318.1 (M+H) + . 1 H NMR(400MHz, CDCl3)δ7.36(dd,J=10.3,4.6Hz,5H),5.25–5.07(m,2H),4.69–4.05(m,2 H),4.05–3.68(m,2H),3.39–3.05(m,2H),2.79–2.44(m,2H),1.48(s,9H),1.15(s,3H).

[0506] Step 8: Preparation of (2S,5R)-2-(cyanomethyl)-5-methylpiperazine-1-carboxylic acid benzyl ester

[0507]

[0508] 2.5 g (6.69 mmol) of 1-benzyl-4-(tert-butyl)(2S,5R)-2-(cyanomethyl)-5-methylpiperazine-1,4-dicarboxylic acid ester was dissolved in dioxane (10 mL). A 1.0 M, 10 mL solution of dioxane in hydrogen chloride was slowly added to the solution with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 2 hours. The reaction mixture was then concentrated to dryness and extracted with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution and saturated brine, dried anhydrous and dried over sodium sulfate, filtered, and concentrated to give a yellow oil (2S,5R)-2-(cyanomethyl)-5-methylpiperazine-1-carboxylic acid benzyl ester (1.8 g, 100% yield). MS: 274.1 (M+H) + .

[0509] Step 9: Preparation of (2S,5R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)-5-methylpiperazine-1-carboxylic acid benzyl ester

[0510]

[0511] Methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3 / step 5, 200 mg, 0.505 mmol) was dissolved in anhydrous dichloromethane (10 mL). Benzyl (2S,5R)-2-(cyanomethyl)-5-methylpiperazine-1-carboxylic acid (138 mg, 0.505 mmol) was slowly added with stirring at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S,5R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)-5-methylpiperazine-1-carboxylic acid benzyl ester (200 mg, 62.6% yield). MS: 633.1 (M+H) + .

[0512] Preparation of intermediate A21 in Example 31

[0513] (2S)-4-(2-chloro-6-((7-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0514]

[0515]

[0516] Step 1: Preparation of (E)-7-chloro-1-(methoxymethylene)-1,2,3,4-tetrahydronaphthalene

[0517]

[0518] Ph3PCH2OMeCl (83.5 g, 243.6 mmol) was dissolved in anhydrous tetrahydrofuran (400 mL), and potassium tert-butoxide (24.9 g, 221.5 mmol) was slowly added with stirring at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour. A tetrahydrofuran solution of 6-chloro-3,4-dihydronaphthyl-1(2H)-one (20.0 g, 110.7 mmol) was added to the above reaction mixture, and the reaction mixture was stirred at 20°C for 12 hours. The reaction was then quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellow solid (E)-7-chloro-1-(methoxymethylene)-1,2,3,4-tetrahydronaphthyl (32 g crude product).

[0519] Step 2: Preparation of 7-chloro-1,2,3,4-tetrahydronaphthalene-1-acetaldehyde

[0520]

[0521] A solution of (E)-7-chloro-1-(methoxymethylene)-1,2,3,4-tetrahydronaphthalene (32.0 g, 153.3 mmol) in formic acid (150 mL) was stirred at 25°C for 16 hours. The pH was then adjusted to 7 with saturated sodium bicarbonate solution, and the mixture was diluted with ethyl acetate for extraction. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow oily substance, 7-chloro-1,2,3,4-tetrahydronaphthalene-1-acetaldehyde (13.7 g, 63.5% yield). 1 H NMR(400MHz,CHLOROFORM-d)δ9.59(d,J=1.9Hz,1H),7.13-7.06(m,2H),7.04-6.98(m,1H),3. 48(t,J=5.5Hz,1H),2.66(t,J=6.3Hz,2H),2.16(m,1H),1.90-1.78(m,1H),1.78-1.60(m,2H).

[0522] Step 3: Preparation of methyl 7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylate

[0523]

[0524] 7-Chloro-1,2,3,4-tetrahydronaphthyl-1-acetaldehyde (13.7 g, 70.4 mmol) was dissolved in acetonitrile (300 mL) and methanol (23.5 g, 731.9 mmol). NIS (47.5 g, 211.1 mmol) and potassium carbonate (229.2 g, 211.1 mmol) were slowly added with stirring at 20°C. After the addition was complete, the reaction mixture was stirred at 25°C for 16 hours. The pH was then adjusted to 9 with saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, methyl 7-chloro-1,2,3,4-tetrahydronaphthyl-1-carboxylate (6.5 g, 41.0% yield). 1 HNMR(400MHz,CHLOROFORM-d)δ7.18(d,J=1.8Hz,1H),7.17-7.13(m,1H),7.07-7.03(m,1H),3.81(t,J=5. 5Hz,1H),3.77-3.73(m,3H),2.87-2.68(m,2H),2.21-2.10(m,1H),2.05-1.92(m,2H),1.83-1.72(m,1H).

[0525] Step 4: Preparation of methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0526]

[0527] Methyl 7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1253 mg, 5.58 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL). LDA (6.97 mmol) was slowly added with stirring at -78°C, and stirring was continued at -78°C for 30 minutes. Then, a tetrahydrofuran solution of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine (intermediate A3-2, 2000 mg, 4.65 mmol) (15 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78°C for 0.5 hours, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a white solid, methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.8 g, 68% yield). MS: 574.1 (M+H) + .

[0528] Step 5: Preparation of methyl 7-chloro-1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0529]

[0530] Methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.8 g, 3.14 mmol) was dissolved in trifluoroacetic acid (5 mL). The reaction mixture was slowly heated to 100 °C under nitrogen protection and stirred in a microwave oven for 1 hour. The reaction mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid methyl 7-chloro-1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1 g, 81% yield). MS: 394.1 (M+H) + .

[0531] Step 6: Preparation of methyl 7-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0532]

[0533] Methyl 7-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (2200 mg, 5.59 mmol) was dissolved in phosphorus oxychloride (15 mL), and DIEA (791 mg, 6.12 mmol) was added with stirring at 0°C. The reaction mixture was slowly heated to 110°C under nitrogen protection and stirred for 6 hours. The reaction mixture was then concentrated, diluted with dichloromethane for extraction, and the solution was poured into a saturated sodium bicarbonate solution and stirred for 1 hour. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The solution was then filtered and concentrated to give methyl 7-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (2.2 g, 91% yield) as a yellow solid. MS: 397.1 (M+H) + .

[0534] Step 7: Preparation of (2S)-4-(2-chloro-6-((7-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0535]

[0536] Methyl 7-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (850 mg, 1.974 mmol) was dissolved in anhydrous dichloromethane (10 mL). (S)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (667 mg, 2.96 mmol) was slowly added with stirring at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour, then the reaction was quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-4-(2-chloro-6-((7-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (0.8 g, 66.7% yield). MS: 620.1 (M+H) + .

[0537] Preparation of intermediate A22 in Example 32

[0538] (2S)-4-(2-chloro-6-((5-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0539]

[0540] The synthesis of intermediate A22 was based on intermediate A21, but was prepared by replacing 7-chloro-3,4-dihydronaphthyl-1(2H)-one with 5-chloro-3,4-dihydronaphthyl-1(2H)-one. MS: 620.1 (M+H) + .

[0541] Preparation of intermediate A23 in Example 33

[0542] (2S)-4-(2-chloro-6-((6-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0543]

[0544] Step 1: Preparation of (E)-6-chloro-1-(methoxymethylene)-1,2,3,4-tetrahydronaphthalene

[0545]

[0546] Ph3PCH2OMeCl (83.5 g, 243.6 mmol) was dissolved in anhydrous tetrahydrofuran (400 mL), and potassium tert-butoxide (24.9 g, 221.5 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour. A tetrahydrofuran solution of 6-chloro-3,4-dihydronaphthyl-1(2H)-one (20.0 g, 110.7 mmol) was added to the above reaction mixture, and the reaction mixture was stirred at 20 °C for 12 hours. The reaction was then quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (E)-6-chloro-1-(methoxymethylene)-1,2,3,4-tetrahydronaphthyl (28.9 g, 75.1% yield). MS: 208.8 (M+H) + .

[0547] Step 2: Preparation of 6-chloro-1,2,3,4-tetrahydronaphthalene-1-acetaldehyde

[0548]

[0549] (E)-6-chloro-1-(methoxymethylene)-1,2,3,4-tetrahydronaphthalene (27.3 g, 130.8 mmol) was dissolved in 1,4-dioxane (270 mL) and water (27 mL). Concentrated hydrochloric acid (12 M, 10.9 mL) was slowly added with stirring at 20°C. After the addition was complete, the reaction mixture was stirred at 60°C for 16 hours. The pH was then adjusted to 9 with saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, 6-chloro-1,2,3,4-tetrahydronaphthalene-1-acetaldehyde (11.2 g, 39.6% yield).

[0550] Step 3: Preparation of methyl 6-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0551]

[0552] 11.4 g (58.6 mmol) of 6-chloro-1,2,3,4-tetrahydronaphthalene-1-acetaldehyde was dissolved in acetonitrile (270 mL) and methanol (18.8 g (585.7 mmol). NIS (39.5 g (175.7 mmol) and potassium carbonate (24.3 g (175.7 mmol)) were slowly added with stirring at 20°C. After the addition was complete, the reaction mixture was stirred at 20°C for 16 hours. The pH was then adjusted to 9 with saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, methyl 6-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (7.6 g, 56.7% yield). 1 H NMR(400MHz,CHLOROFORM-d)δ7.01-7.18(m,3H),3.81(t,J=5.90Hz,1H),3.72-3.75 (m,3H),2.70-2.88(m,2H),2.11-2.22(m,1H),1.91-2.06(m,2H),1.73-1.83(m,1H).

[0553] Step 4: Preparation of methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-6-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0554]

[0555] Methyl 6-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1253 mg, 5.58 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL). LDA (6.97 mmol) was slowly added with stirring at -78°C, and stirring was continued at -78°C for 30 minutes. Then, a tetrahydrofuran solution of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine (intermediate A3-2, 2000 mg, 4.65 mmol) (15 mL) was added to the reaction mixture. After the addition was complete, the reaction mixture was stirred at -78°C for 0.5 hours, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a white solid, methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-6-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.9 g, 73% yield). MS: 574.1 (M+H) + .

[0556] Step 5: Methyl 6-chloro-1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0557]

[0558] Methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-6-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.9 g, 3.3 mmol) was dissolved in trifluoroacetic acid (5 mL). The reaction mixture was slowly heated to 100 °C under nitrogen protection and stirred in a microwave oven for 1 hour. The reaction mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid methyl 6-chloro-1-((2,6-dihydroxy-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.1 g, 82% yield). MS: 394.1 (M+H) + .

[0559] Step 6: Preparation of methyl 6-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0560]

[0561] Methyl 6-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1.1 g, 2.8 mmol) was dissolved in phosphorus oxychloride (15 mL), and DIEA (395 mg, 3.03 mmol) was added with stirring at 0°C. The reaction mixture was slowly heated to 110°C under nitrogen protection and stirred for 6 hours. The reaction mixture was then concentrated, diluted with dichloromethane for extraction, and the solution was poured into a saturated sodium bicarbonate solution and stirred for 1 hour. The organic phase was washed with saturated brine, dried anhydrous, and dried over sodium sulfate. The solution was then filtered and concentrated to give methyl 6-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (1 g, 90% yield) as a yellow solid. MS: 397.1 (M+H) + .

[0562] Step 7: Preparation of (2S)-4-(2-chloro-6-((6-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0563]

[0564] 1 g (2.5 mmol) of methyl 6-chloro-1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid was dissolved in 10 mL of anhydrous dichloromethane. 563 mg (1.78 mmol) of tert-butyl (S)-2-(cyanomethyl)piperazine-1-carboxylic acid was slowly added with stirring at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour. The reaction was then quenched with ice water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-4-(2-chloro-6-((6-chloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (1.1 g, 89.2% yield). MS: 620.1 (M+H) + .

[0565] Preparation of intermediate A24 in Example 34

[0566] (2S)-4-(2-chloro-6-((7-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0567]

[0568] The synthesis of intermediate A24 referenced intermediate A21, and intermediate A24 was prepared by replacing 7-chloro-3,4-dihydronaphthyl-1(2H)-one with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one. MS: 604.1 (M+H) + .

[0569] Preparation of intermediate A25 in Example 35

[0570] (2S)-4-(2-chloro-6-((6-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0571]

[0572] The synthesis of intermediate A25 referenced intermediate A21, and intermediate A25 was prepared by replacing 7-chloro-3,4-dihydronaphthyl-1(2H)-one with 6-fluoro-3,4-dihydronaphthyl-1(2H)-one. MS: 604.1 (M+H) + .

[0573] Preparation of intermediate A26 in Example 36

[0574] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-7-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0575]

[0576] The synthesis of intermediate A26 referenced intermediate A21, and intermediate A26 was prepared by replacing 7-chloro-3,4-dihydronaphthyl-1(2H)-one with 7-methyl-3,4-dihydronaphthyl-1(2H)-one. MS: 600.1 (M+H) + .

[0577] Preparation of intermediates A27 and A28 in Example 37

[0578] (S)-1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (intermediate A27) and (R)-1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (intermediate A28)

[0579]

[0580] Methyl 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (44 g) was chirally resolved by SFC (column type: CHIRAL ART Cellulose-SB, 5 cm × 25 cm, 5 μm column; mobile phase: ethanol in n-hexane) to give the first component peak intermediate A27 (retention time 3.86 min, 20 g) and the second component peak intermediate A28 (retention time 4.11 min, 19 g), both pale yellow solids. MS: 540.1 (M+H) + Intermediate A27 was determined to be of (S)-configuration by X-ray crystal structure analysis, as shown in the following results. Figure 1 As shown.

[0581] Preparation of intermediate A29 in Example 38

[0582] (S)-4-(2-chloro-6-(((S)-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0583]

[0584] The synthesis of intermediate A29 referenced intermediate A3, and intermediate A29 was prepared by replacing 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (intermediate A27) with (S)-1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (intermediate A3-3). MS: 586.1 (M+H) + .

[0585] Preparation of intermediate A30 in Example 39

[0586] (S)-4-(2-chloro-6-(((R)-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0587]

[0588] The synthesis of intermediate A30 referenced intermediate A3, and intermediate A30 was prepared by replacing 1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (intermediate A28) with (R)-1-(R)-1-((2,6-bis(benzyloxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (intermediate A3-3). MS: 586.1 (M+H) + .

[0589] Preparation of intermediate A31 in Example 40

[0590] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-6-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0591]

[0592] The synthesis of intermediate A31 referenced intermediate A21, and intermediate A31 was prepared by replacing 7-chloro-3,4-dihydronaphthyl-1(2H)-one with 6-methyl-3,4-dihydronaphthyl-1(2H)-one. MS: 600.1 (M+H) + .

[0593] Preparation of intermediate A32 in Example 41

[0594] (2S)-4-(2-chloro-6-((5-chloro-1-(methoxycarbonyl)-7-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0595]

[0596]

[0597] Step 1: 4-(2-chloro-4-methylphenyl)but-3-yn-1-ol

[0598]

[0599] 2-Chloro-1-iodo-4-toluene (40 g, 158 mmol) was dissolved in tetrahydrofuran (300 mL). Butyronitrile (167 g, 2376 mmol), CuI (0.603 g, 3.17 mmol), Pd(PPh3)2Cl2 (3.34 g, 4.75 mmol), and TEA (40.1 g, 396 mmol) were slowly added with stirring at 0 L. After the addition was complete, the reaction mixture was stirred for 12 hours at 60 drops, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil, 4-(2-chloro-4-methylphenyl)butyronitrile (26 g, 84% yield). MS: 195.2 (M+H) + .

[0600] Step 2: Preparation of 4-(2-chloro-4-methylphenyl)butane-1-ol

[0601]

[0602] 4-(2-chloro-4-methylphenyl)butan-1-ol (9 g, 45.3 mmol) was dissolved in ethyl acetate (200 mL), and PtO2 (1 g) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature under hydrogen atmosphere for 12 hours, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a yellow oily substance, 4-(2-chloro-4-methylphenyl)butan-1-ol (9 g, 98% yield). MS: 199.2 (M+H) + .

[0603] Step 3: Preparation of 4-(2-chloro-4-methylphenyl)butyric acid

[0604]

[0605] 4-(2-chloro-4-methylphenyl)butan-1-ol (8 g, 40.3 mmol) was dissolved in acetone (30 mL), and Jones' reagent (10.41 g, 81 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 2 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid 4-(2-chloro-4-methylphenyl)butyric acid (3.8 g, 44.4% yield). MS: 213.2 (M+H) + .

[0606] Step 4: Preparation of 5-chloro-7-methyl-3,4-dihydronaphth-1(2H)-one

[0607]

[0608] 4-(2-chloro-4-methylphenyl)butyric acid (3.8 g, 17.87 mmol) was dissolved in trifluoromethanesulfonic acid (10 mL), and the reaction was stirred at 0 °C for 3 h. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, 5-chloro-7-methyl-3,4-dihydronaphthyl-1(2H)-one (2.2 g, 63.3% yield). MS: 195.2 (M+H) + .

[0609] Step 5: Preparation of (E)-5-chloro-1-(methoxymethylene)-7-methyl-1,2,3,4-tetrahydronaphthalene

[0610]

[0611] The synthesis of intermediate A32-5 is based on intermediate A21 / step 1, and intermediate A32-5 is prepared by using 5-chloro-7-methyl-3,4-dihydronaphthyl-1(2H)-one (intermediate A32-4) instead of 7-chloro-3,4-dihydronaphthyl-1(2H)-one.

[0612] Step 6: Preparation of 5-chloro-7-methyl-1,2,3,4-tetrahydronaphthalene-1-carboxaldehyde

[0613]

[0614] (E)-5-chloro-1-(methoxymethylene)-7-methyl-1,2,3,4-tetrahydronaphthalene (2.4 g, 10.78 mmol) was dissolved in dichloromethane (20 mL), and BBr3 (43.1 mmol) was slowly added with stirring at -40 °C. After the addition was complete, the reaction mixture was stirred at -40 °C for 2 hours, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, 5-chloro-7-methyl-1,2,3,4-tetrahydronaphthalene-1-carboxaldehyde (700 mg, 31.3% yield).

[0615] Step 7: Preparation of methyl 5-chloro-7-methyl-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0616]

[0617] The synthesis of intermediate A32-7 follows the procedure of intermediate A21 / step 3, using 5-chloro-7-methyl-1,2,3,4-tetrahydronaphthalene-1-carboxaldehyde (intermediate A32-6) instead of 7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxaldehyde (intermediate A21 / step 2). MS: 239.2 (M+H) + .

[0618] Step 8: Preparation of (2S)-4-(2-chloro-6-((5-chloro-1-(methoxycarbonyl)-7-methyl-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0619]

[0620] The synthesis of intermediate A32 referenced intermediate A21, and intermediate A32 was prepared by replacing methyl 5-chloro-7-methyl-1,2,3,4-tetrahydronaphthalene-1-carboxylate (intermediate A32-7) with methyl 7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxylate (intermediate A21 / step 3). MS: 634.2 (M+H) + .

[0621] Preparation of intermediate A33 in Example 42

[0622] (2S)-4-(2-chloro-6-((5-chloro-1-(methoxycarbonyl)-6-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0623]

[0624]

[0625] The synthesis of intermediate A33 referenced intermediate A32, but was prepared by replacing 2-chloro-1-iodo-3-toluene with 2-chloro-1-iodo-4-toluene. MS: 634.2 (M+H) + .

[0626] Preparation of intermediate A34 in Example 43

[0627] (2S)-4-(2-chloro-6-((5,6-difluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0628]

[0629] Step 1: Preparation of 5,6-difluoro-3,4-dihydronaphthyl-1(2H)-one (intermediate A34-4)

[0630] The synthesis of intermediate A34-4 referenced intermediate A32-4, and was prepared by replacing 2-chloro-1-iodo-4-toluene with 1,2-difluoro-3-iodobenzene. MS: 183.2 (M+H) + .

[0631] Step 2: Preparation of (2S)-4-(2-chloro-6-((5,6-difluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A34)

[0632] The synthesis of intermediate A34 referenced intermediate A23, but was prepared by replacing 6-chloro-3,4-dihydronaphthyl-1(2H)-one with 5,6-difluoro-3,4-dihydronaphthyl-1(2H)-one (intermediate A34-4). MS: 622.2 (M+H) + .

[0633] Preparation of intermediate A35 in Example 44

[0634] (2S)-4-(2-chloro-6-((5-chloro-6-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0635]

[0636] Step 1: Preparation of 5-chloro-6-fluoro-3,4-dihydronaphth-1(2H)-one (Intermediate A35-4)

[0637]

[0638] The synthesis of intermediate A35-4 referenced intermediate A32-4, but was prepared by replacing 2-chloro-1-fluoro-3-iodobenzene with 2-chloro-1-iodo-4-toluene. MS: 199.2 (M+H) + .

[0639] Step 2: Preparation of 5-chloro-6-fluoro-1-methylene-1,2,3,4-tetrahydronaphthalene (intermediate A35-5)

[0640]

[0641] Methyltriphenylphosphonium bromide (27.3 g, 77 mmol) was dissolved in tetrahydrofuran (180 mL), and potassium tert-butoxide (8.59 g, 77 mmol) was slowly added with stirring at 0 °C for 1 hour. Then, a tetrahydrofuran solution (50 mL) for the preparation of 5-chloro-6-fluoro-3,4-dihydronaphthyl-1(2H)-one (intermediate A35-4, 7.6 g, 38.3 mmol) was added to the above reaction solution at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 12 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a colorless oil, 5-chloro-6-fluoro-1-methylene-1,2,3,4-tetrahydronaphthyl (intermediate A35-5, 6.4 g, 85% yield).

[0642] Step 3: Preparation of (5-chloro-6-fluoro-1,2,3,4-tetrahydronaphth-1-yl)methanol (intermediate A35-6)

[0643]

[0644] 5-Chloro-6-fluoro-1-methylene-1,2,3,4-tetrahydronaphthalene (intermediate A35-5, 6.4 g, 32.5 mmol) was dissolved in tetrahydrofuran (80 mL). A borane tetrahydrofuran solution (65.1 mmol) was slowly added with stirring at 0 °C, and the mixture was stirred at 0 °C for 1 hour. Then, NaOH (98 mmol) and H₂O₂ (521 mmol) were added to the reaction mixture at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 12 hours, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a colorless oil (5-chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-yl)methanol (intermediate A35-6, 6.0 g, 86% yield).

[0645] Step 4: 5-Chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxaldehyde (intermediate A35-7)

[0646]

[0647] (5-Chloro-6-fluoro-1,2,3,4-tetrahydronaphth-1-yl)methanol (intermediate A35-6, 6.0 g, 28.0 mmol) was dissolved in dichloromethane (80 mL). Sodium bicarbonate (4.70 g, 55.9 mmol) and dess-martinperiodinane (23.71 g, 55.9 mmol) were slowly added with stirring at 0 °C, and the mixture was stirred at 0 °C for 1 hour. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous, and dried over sodium sulfate. The mixture was then filtered and concentrated to give a colorless oily substance, 5-chloro-6-fluoro-1,2,3,4-tetrahydronaphth-1-carboxaldehyde (intermediate A35-7, 6.0 g crude).

[0648] Step 5: Preparation of methyl 5-chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A35-8)

[0649]

[0650] The synthesis of intermediate A35-8 follows the procedure of intermediate A21 / step 3, but instead of 7-chloro-1,2,3,4-tetrahydronaphthalene-1-carboxaldehyde (intermediate A35-7), intermediate A35-8 was prepared. MS: 243.2 (M+H) + .

[0651] Step 6: (2S)-4-(2-chloro-6-((5-chloro-6-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A35)

[0652]

[0653] The synthesis of intermediate A35 referenced intermediate A32, and intermediate A35 was prepared by replacing methyl 5-chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylate (intermediate A35-8) with methyl 5-chloro-7-methyl-1,2,3,4-tetrahydronaphthalene-1-carboxylate (intermediate A32-7). MS: 638.1 (M+H) + .

[0654] Preparation of intermediate A36 in Example 45

[0655] (2S)-4-(2-chloro-6-((5-chloro-7-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0656]

[0657] Step 1: Preparation of (E)-4-(2-chloro-4-fluorophenyl)but-3-enoic acid

[0658]

[0659] (2-Carboxyethyl)triphenylphosphine bromide (79 g, 189 mmol) was dissolved in tetrahydrofuran (300 mL), and potassium tert-butoxide (53 g, 473 mmol) was slowly added with stirring at 0 °C for 10 min. Then, a solution of 2-chloro-4-fluorobenzaldehyde (25 g, 158 mmol) in tetrahydrofuran (50 mL) was added to the above reaction solution at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 5 h, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil (E)-4-(2-chloro-4-fluorophenyl)but-3-enoic acid (20 g, 59.2% yield). MS: 215.2 (M+H) + .

[0660] Step 2: Preparation of 4-(2-chloro-4-fluorophenyl)butyric acid

[0661]

[0662] (E)-4-(2-chloro-4-fluorophenyl)but-3-enoic acid (15 g, 69.7 mmol) was dissolved in ethyl acetate (200 mL), and PtO2 (1 g) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature under hydrogen atmosphere for 12 hours, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a yellow oily substance, 4-(2-chloro-4-fluorophenyl)butyric acid (13.6 g, 90% yield). MS: 217.2 (M+H) + .

[0663] Step 3: Preparation of 5-chloro-7-fluoro-3,4-dihydronaphthyl-1(2H)-one

[0664]

[0665] The synthesis of intermediate A36-3 referenced intermediate A32-4, and intermediate A36-3 was prepared by replacing 4-(2-chloro-4-fluorophenyl)butyric acid (intermediate A36-2) with 4-(2-chloro-4-methylphenyl)butyric acid (intermediate A32-3). MS: 199.1 (M+H) + .

[0666] Step 4: Preparation of (2S)-4-(2-chloro-6-((5-chloro-7-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0667]

[0668] The synthesis of intermediate A36 referenced intermediate A23, but was prepared by replacing 6-chloro-3,4-dihydronaphthyl-1(2H)-one with 5-chloro-7-fluoro-3,4-dihydronaphthyl-1(2H)-one (intermediate A36-3). MS: 638.2 (M+H) + .

[0669] Preparation of intermediate A37 in Example 46

[0670] (2S)-4-(2-chloro-6-((5,7-dichloro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0671]

[0672] The synthesis of intermediate A37 referenced intermediate A36, but was obtained by replacing 2-chloro-4-fluorobenzaldehyde with 2,4-dichlorobenzaldehyde. MS: 654.1 (M+H) + .

[0673] Preparation of intermediate A38 in Example 47

[0674] (2S)-4-(2-chloro-6-((5-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0675]

[0676]

[0677] The synthesis of intermediate A38 references intermediate A34, and intermediate A38 is obtained by replacing 1,2-difluoro-3-iodobenzene with 1-fluoro-2-iodobenzene. MS: 604.1 (M+H) + .

[0678] Preparation of intermediate A39 in Example 48

[0679] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-5-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0680]

[0681] The synthesis of intermediate A39 references intermediate A34, and intermediate A39 is obtained by replacing 1,2-difluoro-3-iodobenzene with 1-iodo-2-toluene. MS: 600.1 (M+H) + .

[0682] Preparation of intermediate A40 in Example 49

[0683] (2S)-4-(2-chloro-6-((7-fluoro-1-(methoxycarbonyl)-5-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0684]

[0685] The synthesis of intermediate A40 references intermediate A34, but is obtained by replacing 1,2-difluoro-3-iodobenzene with 1-bromo-4-fluoro-2-toluene. MS: 618.1 (M+H) + .

[0686] Preparation of intermediate A41 in Example 50

[0687] (2S)-4-(2-chloro-6-((5-fluoro-1-(methoxycarbonyl)-6-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0688]

[0689] The synthesis of intermediate A41 references intermediate A34, but is obtained by replacing 1,2-difluoro-3-iodobenzene with 1-bromo-2-fluoro-3-toluene. MS: 618.1 (M+H) + .

[0690] Preparation of intermediate A42 in Example 51

[0691] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-5-(trifluoromethoxy)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0692]

[0693] Step 1: Preparation of 5-(trifluoromethoxy)-3,4-dihydronaphthyl-1(2H)-one

[0694]

[0695] The synthesis of intermediate A42-3 was based on intermediate A36-3, but was prepared by replacing 2-chloro-4-fluorobenzaldehyde with 2-trifluoromethoxybenzaldehyde. MS: 231.1 (M+H) + .

[0696] Step 2: Preparation of (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-5-(trifluoromethoxy)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0697]

[0698] The synthesis of intermediate A42 referenced intermediate A35, and intermediate A42 was prepared by replacing 5-chloro-6-fluoro-3,4-dihydronaphthyl-1(2H)-one (intermediate 42-3) with 5-(trifluoromethoxy)-3,4-dihydronaphthyl-1(2H)-one (intermediate A35-4). MS: 670.1 (M+H) + .

[0699] Preparation of intermediate A43 in Example 52

[0700] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0701]

[0702] The synthesis of intermediate A43 references intermediate A34, and intermediate A43 is obtained by replacing 1,2-difluoro-3-iodobenzene with 1-iodo-2-(trifluoromethyl)benzene. MS: 654.1 (M+H) + .

[0703] Preparation of intermediate A51 in Example 53

[0704] (2S)-4-(2-chloro-6-((6-chloro-5-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0705]

[0706] The synthesis of intermediate A51 references intermediate A32, but is obtained by replacing 2-chloro-1-iod-4-toluene with 1-chloro-2-fluoro-3-iodobenzene. MS: 638.2 (M+H) + .

[0707] Preparation of intermediate A44 in Example 54

[0708] (2S)-4-(2-chloro-6-((5,7-difluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0709]

[0710] The synthesis of intermediate A44 referenced intermediate A35, and intermediate A44 was prepared by replacing 2-chloro-1-fluoro-3-iodobenzene with 2,4-difluoro-1-iodobenzene. MS: 622.1 (M+H) + .

[0711] Preparation of intermediate A46 in Example 55

[0712] (2S)-4-(2-chloro-6-((5-fluoro-1-(methoxycarbonyl)-7-methyl-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid

[0713]

[0714] The synthesis of intermediate A46 referenced intermediate A35, and was prepared by replacing 2-chloro-1-fluoro-3-iodobenzene with 2-fluoro-1-iodo-4-toluene. MS: 618.1 (M+H) + .

[0715] Preparation of intermediate A47 in Example 56

[0716] (2S)-4-(2-chloro-6-((7-chloro-1-(methoxycarbonyl)-5-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0717]

[0718] The synthesis of intermediate A47 referenced intermediate A34, and was prepared by replacing 1,2-difluoro-3-iodobenzene with 4-chloro-1-iodo-2-toluene. MS: 634.1 (M+H) + .

[0719] Preparation of intermediate A48 in Example 57

[0720] (2S)-4-(2-chloro-6-((6-fluoro-1-(methoxycarbonyl)-5-methyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0721]

[0722] The synthesis of intermediate A48 referenced intermediate A34, and was prepared by replacing 1,2-difluoro-3-iodobenzene with 1-fluoro-3-iodo-2-toluene. MS: 618.1 (M+H) + .

[0723] Preparation of intermediate A45 in Example 58

[0724] (2S)-4-(6-((5-bromo-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-chloro-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0725]

[0726] The synthesis of intermediate A45 referenced intermediate A35, and intermediate A45 was prepared by replacing 5-chloro-6-fluoro-3,4-dihydronaphthyl-1(2H)-one with 5-bromo-3,4-dihydronaphthyl-1(2H)-one (intermediate A35-4). MS: 618.1 (M+H) + .

[0727] Preparation of intermediate A49 in Example 59

[0728] (2S)-4-(2-chloro-6-((4-chloro-1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0729]

[0730] The synthesis of intermediate A49 referenced intermediate A34, and intermediate A49 was prepared by replacing 5,6-difluoro-3,4-dihydronaphthyl-1(2H)-one with 4-chloro-2,3-dihydro-1H-inden-1-one (intermediate A34-4). MS: 605.4 (M+H) + .

[0731] Preparation of intermediate A52 in Example 60

[0732] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-4,4-dimethyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0733]

[0734] Step 1: Preparation of intermediate A52-1

[0735] 2-Phenylacetonitrile (5.00 g, 42.7 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), and sodium hydrogen (1.88 g, 47.0 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 15 °C for 0.5 h. Then, 1-bromo-3-methyl-but-2-ene was added to the above reaction solution, and the reaction mixture was stirred at 15 °C for 16 h. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oily intermediate A52-1 (7.4 g). 1H NMR (400MHz, CDCl3) δ7.42-7.33(m,5H),5.25-5.18(m,1H),3.79(dd,J=6.5,8.1Hz,1H),2.66-2.54(m,2H),1.77-1.72(m,3H),1.57(s,3H).

[0736] Step 2: Preparation of 4,4-dimethyl-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A52-2)

[0737] Intermediate A52-1 (7.4 g, 39.9 mmol) was slowly added to a sulfuric acid solution (22.9 g, 139.8 mmol, 12.4 mL, 60% purity) with stirring at 60 °C. After the addition was complete, the reaction mixture was stirred at 130 °C for 16 hours. The reaction was then quenched in ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oil, 4,4-dimethyl-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (6.6 g). 1 HNMR(400MHz, CDCl3)δ11.45(br s, 1H), 7.30 (d, J=7.9Hz, 1H), 7.19-7.03 (m, 3H), 3.79-3.68 (m, 1H) 2.21-1.91 (m,2H),1.86-1.72(m,1H),1.55-1.48(m,1H),1.26(s,3H),1.22-1.15(m,3H).

[0738] Step 3: Preparation of methyl 4,4-dimethyl-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A52-3)

[0739] 6.6 g (32.3 mmol) of 4,4-dimethyl-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid was dissolved in 3.0 M methanol hydrochloride (30 mL). The reaction mixture was stirred at 70 °C for 16 h. The mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give 5.9 g of methyl 4,4-dimethyl-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid. 1H NMR (400MHz, CDCl3) δ7.40 (d, J = 7.8Hz, 1H), 7.28-7.21 (m, 1H), 7.18-7.11 (m, 2H), 3.88-3.81 (m, 1H), 3.77 (s ,3H),2.23-2.13(m,1H),2.12-2.01(m,1H),1.87-1.82(m,1H),1.65-1.53(m,1H),1.36(s,3H),1.30(s,3H).

[0740] Step 4: Preparation of (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-4,4-dimethyl-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0741] The synthesis of intermediate A52 referenced intermediate A35, and intermediate A52 was prepared by replacing methyl 4,4-dimethyl-1,2,3,4-tetrahydronaphthalene-1-carboxylate (intermediate A52-3) with methyl 5-chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylate (intermediate A35-8). MS: 614.1 (M+H) + .

[0742] Preparation of intermediate C6 in Example 61

[0743] (S)-(4,4-Difluoro-1-methylpyrrolidin-2-yl)methanol

[0744]

[0745] The synthesis of intermediate C6 referenced intermediate C3, and was prepared by replacing 1-(tert-butyl)-2-methyl(S)-4,4-difluoropyrrolidine-1,2-dicarboxylic acid ester with 1-(tert-butyl)-2-methyl(2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylic acid ester. MS: 152.1 (M+H) + .

[0746] Preparation of intermediate A53 in Example 62

[0747] (2S)-4-(2-chloro-6-((5-chloro-1-(methoxycarbonyl)isochloro-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0748]

[0749] Step 1: Preparation of intermediate A53-1

[0750] 10 g (63.9 mmol) of 2-(2-chlorophenyl)ethane-1-ol was dissolved in 40 mL of TFA, and 8.82 g (96 mmol) of 2,2-dihydroxyacetic acid was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 100 °C for 48 hours. The mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was recrystallized from dichloromethane to give a white solid intermediate A53-1 (3.2 g, 23.5%).

[0751] Step 2: Preparation of intermediate A53-2

[0752] Intermediate A53-1 (3.5 g, 16.46 mmol) was dissolved in 10 mL of methanol, and thionyl chloride (9.79 g, 82 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 2 hours. The mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a colorless oily intermediate A53-2 (2.6 g, 69.7%). MS: 227.1 (M+H) + .

[0753] Step 3: Preparation of (2S)-4-(2-chloro-6-((5-chloro-1-(methoxycarbonyl)isocyano-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0754] The synthesis of intermediate A53 referenced intermediate A35, and intermediate A53 was prepared by replacing methyl 5-chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A35-8) with intermediate A53-2. MS: 621.1 (M+H) + .

[0755] Preparation of intermediate C7 in Example 63

[0756] ((2S,4R,5S)-4-fluoro-1,5-dimethylpyrrolid-2-yl)methanol

[0757]

[0758] The synthesis of intermediate C7 referenced intermediate C3, and was prepared by replacing 1-(tert-butyl)-2-methyl(2S,4R,5S)-4-fluoro-5-methylpyrrolidine-1,2-dicarboxylic acid ester with 1-(tert-butyl)-2-methyl(2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylic acid ester. MS: 148.1 (M+H) + .

[0759] Preparation of intermediate A54 in Example 64

[0760] (2S)-4-(2-chloro-6-((4-chloro-6-fluoro-1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0761]

[0762] Step 1: Preparation of 4-chloro-6-fluoro-2,3-dihydro-1H-inden-1-one (Intermediate A54-1)

[0763] The synthesis of intermediate A54-1 is based on intermediate A32-4. Intermediate A54-1 was prepared by replacing 4-(2-chloro-4-fluorophenyl)propionic acid (intermediate A32-3) with 3-(2-chloro-4-fluorophenyl)propionic acid.

[0764] Step 2: Preparation of (2S)-4-(2-chloro-6-((4-chloro-6-fluoro-1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A54)

[0765] The synthesis of intermediate A54 referenced intermediate A34, and intermediate A54 was prepared by replacing 5,6-difluoro-3,4-dihydronaphthyl-1(2H)-one (intermediate A34-4) with 4-chloro-6-fluoro-2,3-dihydro-1H-inden-1-one (intermediate A54-1). MS: 623.4 (M+H) + .

[0766] Preparation of intermediate A55 in Example 65

[0767] (2S)-4-(2-chloro-6-((5-chloro-1-(ethoxycarbonyl)-7-fluoroisocyano-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0768]

[0769] Step 1: Preparation of ethyl 2-(2-chloro-4-fluorophenylethoxy)-2-ethoxyacetate (intermediate A55-1)

[0770] 2-(2-chloro-4-fluorophenyl)ethane-1-ol (25 g, 143 mmol) and ethyl 2,2-diethoxyethyl acetate (50.5 g, 286 mmol) were dissolved in 300 mL of DCE solution. Boron trifluoride diethyl ether (4.37 g, 14.32 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 13 hours. The mixture was then concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a colorless oily intermediate A55-1 (20 g, 45.8%).

[0771] Step 2: Preparation of intermediate A55-2

[0772] Ethyl 2-(2-chloro-4-fluorophenylethoxy)-2-ethoxyacetate (intermediate A55-1, 19 g, 62.3 mmol) was dissolved in 100 mL of DCE solution, and Eaton reagent (14.84 g, 62.3 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 13 hours. The reaction mixture was then added to ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a colorless oily intermediate A55-2 (9.6 g, 59.5%). MS: 259.1 (M+H) + .

[0773] Step 3: Preparation of (2S)-4-(2-chloro-6-((5-chloro-1-(ethoxycarbonyl)-7-fluoroisocyanoamine-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0774] The synthesis of intermediate A55 referenced intermediate A35, and intermediate A55 was prepared by replacing methyl 5-chloro-6-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A35-8) with intermediate A55-2. MS: 653.4 (M+H) + .

[0775] Preparation of intermediate A56 in Example 66

[0776] (2S)-4-(2-chloro-6-((5-chloro-1-(ethoxycarbonyl)-8-fluoroisocyano-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0777]

[0778] The synthesis of intermediate A56 referenced intermediate A53, but was prepared by replacing 2-(2-chloro-5-fluorophenyl)ethane-1-ol with 2-(2-chlorophenyl)ethane-1-ol. MS: 639.4 (M+H) + .

[0779] Preparation of intermediate A57 in Example 67

[0780] methyl 5-chloro-8-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0781]

[0782] The synthesis of intermediate A57 referenced intermediate A34-7, and was prepared by replacing 1,2-difluoro-3-iodobenzene with 2-bromo-1-chloro-4-fluorobenzene. MS: 242.4 (M+H) + .

[0783] Preparation of intermediate A58 in Example 68

[0784] (2S)-4-(5-((tert-butoxycarbonyl)amino)-6-((5-chloro-8-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0785]

[0786] Step 1: Preparation of methyl 2,6-bis(benzyloxy)-5-nitropyrimidine-4-yl)acetate (intermediate A58-2)

[0787] 10 g (23.24 mmol) of 2,4-bis(benzyloxy)-6-(bromomethyl)-5-nitropyrimidine was dissolved in 50 mL of DMF. Potassium acetate (3.42 g, 34.9 mmol) was added with stirring at 0 °C, and the mixture was stirred at 20 °C for 2 hours. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous, and dried over sodium sulfate. The mixture was then filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, methyl 2,6-bis(benzyloxy)-5-nitropyrimidine-4-yl)acetate (intermediate A58-2, 9.0 g, 95%). MS: 410.2 (M+H) + .

[0788] Step 2: Preparation of (2,6-dihydroxy-5-nitropyrimidine-4-yl)methyl acetate (intermediate A58-3)

[0789] 2,6-bis(benzyloxy)-5-nitropyrimidine-4-yl)methyl acetate (9 g, 21.98 mmol) was dissolved in trifluoroacetic acid (50 mL). The reaction mixture was stirred at 50 °C for 12 h under nitrogen protection, then the reaction mixture was concentrated and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid (2,6-dihydroxy-5-nitropyrimidine-4-yl)methyl acetate (intermediate A58-3, 4.2 g, 83%). MS: 230.1 (M+H) + .

[0790] Step 3: Preparation of (2,6-dichloro-5-nitropyrimidin-4-yl)methyl acetate (intermediate A58-4)

[0791] Methyl (2,6-dihydroxy-5-nitropyrimidin-4-yl)acetate (intermediate A58-3, 4.2 g, 18.33 mmol) was dissolved in toluene (50 mL). Phosphorus oxychloride (28.1 g, 183 mmol) and DIEA (7.11 g, 55.0 mmol) were added with stirring at 0°C. The reaction mixture was slowly heated to 110°C under nitrogen protection and stirred for 2 hours. The reaction mixture was then concentrated, diluted with dichloromethane for extraction, and the solution was poured into a saturated sodium bicarbonate solution and stirred for 1 hour. The organic phase was washed with saturated brine, dried anhydrously, and dried over sodium sulfate. The solution was then filtered and concentrated to give a yellow solid of methyl (2,6-dichloro-5-nitropyrimidin-4-yl)acetate (intermediate A58-4, 3.1 g, 63.6%). MS: 266.1 (M+H) + .

[0792] Step 4: Preparation of (S)-4-(6-(acetoxymethyl)-2-chloro-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-5)

[0793] Methyl (2,6-dichloro-5-nitropyrimidin-4-yl)acetate (intermediate A58-4, 3.1 g, 11.65 mmol) was dissolved in DCM (50 mL). (S)-2-(piperazin-2-yl)acetonitrile (1.459 g, 11.65 mmol) was added to the reaction mixture at 0°C with stirring, and the mixture was stirred at 20°C for 1 hour. Then, Boc2O (5.09 g, 23.31 mmol) and DIEA (1.506 g, 11.65 mmol) were added to the above reaction mixture at 0°C, and the mixture was stirred at 50°C for 1 hour. The reaction mixture was poured into ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (S)-4-(6-(acetoxymethyl)-2-chloro-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-5, 3.3 g, 62.3%). MS: 455.2 (M+H) + .

[0794] Step 5: Preparation of (S)-4-(6-(acetoxymethyl)-2-(methylthio)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-6)

[0795] A solution of (S)-4-(6-(acetoxymethyl)-2-chloro-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-5, 1.6 g, 3.52 mmol) and CH3SNa (0.493 g, 7.04 mmol) in THF (10 mL) and water (5 mL) was stirred at 20°C for 2 h. The reaction mixture was poured into ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid (S)-4-(6-(acetoxymethyl)-2-(methylthio)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-6, 1.3 g, 79%). MS: 467.2 (M+H) + .

[0796] Step 6: Preparation of (S)-4-(6-(acetoxymethyl)-5-amino-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-7)

[0797] (S)-4-(6-(acetoxymethyl)-2-(methylthio)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-6, 1.3 g, 2.79 mmol) was dissolved in ethanol (15 mL). Iron powder (0.778 g, 13.93 mmol) and ammonium chloride solution (3 mL) were added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 85 °C for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (S)-4-(6-(acetoxymethyl)-5-amino-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-7, 1.0 g, 82%). MS: 437.2 (M+H) + .

[0798] Step 7: Preparation of (S)-4-(6-(acetoxymethyl)-5-((tert-butyloxycarbonyl)amino)-2-(methylthio)pyrimidin-4-yl)-2-(cyano)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-8)

[0799] A mixture of (S)-4-(6-(acetoxymethyl)-5-amino-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-7, 1.0 g, 2.291 mmol), Boc2O (5 mL), and DIEA (0.592 g, 4.58 mmol) was stirred at 80 °C for 12 h, then the reaction was quenched with ice water and extracted with DCM. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (S)-4-(6-(acetoxymethyl)-5-((tert-butyloxycarbonyl)amino)-2-(methylthio)pyrimidin-4-yl)-2-(cyano)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-8, 950 mg, 77%). MS: 537.3 (M+H) + .

[0800] Step 8: Preparation of (S)-4-(5-((tert-Butoxycarbonyl)amino)-6-(hydroxymethyl)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid (intermediate A58-9)

[0801] A mixture of (S)-4-(6-(acetoxymethyl)-5-((tert-butyloxycarbonyl)amino)-2-(methylthio)pyrimidin-4-yl)-2-(cyano)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-8, 950 mg, 1.77 mmol), lithium hydroxide (212 mg, 8.85 mmol), THF (5 mL), and water (5 mL) was stirred at 20 °C for 2 h. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow solid (S)-4-(5-((tert-butyloxycarbonyl)amino)-6-(hydroxymethyl)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid (intermediate A58-9, 650 mg, 74.2%). MS: 495.3 (M+H) + .

[0802] Step 9: Preparation of (S)-4-(6-(bromomethyl)-5-((tert-butyloxycarbonyl)amino)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-10)

[0803] (S)-4-(5-((tert-Butoxycarbonyl)amino)-6-(hydroxymethyl)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazin-1-carboxylic acid (intermediate A58-9, 650 mg, 1.314 mmol) was dissolved in DCM (10 mL). Ph3P (517 mg, 1.971 mmol) and CBr4 (654 mg, 1.971 mmol) were added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 2 hours, then the reaction was quenched with ice water and extracted with DCM. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (S)-4-(6-(bromomethyl)-5-((tert-butyloxycarbonyl)amino)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-10, 350 mg, 47.8%). MS: 557.2 & 559.2 (M+H) + .

[0804] Step 10: Preparation of (2S)-4-(5-((tert-Butoxycarbonyl)amino)-6-((5-chloro-8-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58)

[0805] Methyl 5-chloro-8-fluoro-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (305 mg, 1.256 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL). LDA (1.25 mmol) was slowly added with stirring at -78 °C, and stirring continued at -20 °C for 60 min. Then, (S)-4-(6-(bromomethyl)-5-((tert-butoxycarbonyl)amino)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A58-10, 350 mg, 0.628 mmol) was added to the above reaction solution at -78 °C. After the addition was complete, the reaction mixture was stirred at -78 °C for 1 h, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-4-(5-((tert-butoxycarbonyl)amino)-6-((5-chloro-8-fluoro-1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(methylthio)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A 58, 220 mg, 48.7%). MS: 719.4 (M+H) + .

[0806] Preparation of intermediate A59 in Example 69

[0807] (2S)-4-(2-chloro-6-((4-chloro-5-fluoro-1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[0808]

[0809] The synthesis of intermediate A59 referenced intermediate A54, and intermediate A59 was prepared by replacing 3-(2-chloro-3-fluorophenyl)propionic acid with 3-(2-chloro-4-fluorophenyl)propionic acid. MS: 623.4 (M+H) + .

[0810] Preparation of intermediate C8 in Example 70

[0811] ((2R)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol

[0812]

[0813] Step 1: Preparation of intermediate C8-1

[0814] 20 g (81 mmol) of 1-(tert-butyl)-2-methyl(2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylate and HMPA (23.19 g, 129 mmol) were dissolved in anhydrous tetrahydrofuran (400 mL). LiHMDS (129 mL) was slowly added with stirring at -70 °C, and stirring was continued at -70 °C for 60 min. Then, a solution of 1-bromo-3-chloropropane (63.7 g, 404 mmol) in tetrahydrofuran (100 mL) was added to the above reaction solution at -70 °C. After the addition was complete, the reaction mixture was stirred at -70 °C for 2 h, and then at room temperature for 2 h. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oily intermediate C8-1 (16 g, 61.1%). MS:323.1&267.1(M+H) + .

[0815] Step 2: Preparation of intermediate C8-2

[0816] Intermediate C8-1 (52 g, 161 mmol) was dissolved in acetonitrile (100 mL). A 1.0 M, 200 mL solution of dioxane (hydrogen chloride) was slowly added with stirring at 0 °C. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated to dryness, diluted with ethyl acetate, and extracted. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give intermediate C8-2 (36 g, 100%) as a yellow oil. MS: 224.1 (M+H) + .

[0817] Step 3: Preparation of intermediate C8-3

[0818] Intermediate C8-2 (36 g, 161 mmol) was slowly added to acetonitrile (50 mL) with stirring at 0 °C, along with NaHCO3 (67.6 g, 805 mmol) and potassium iodide (2.67 g, 16.09 mmol). The reaction mixture was then stirred at 50 °C for 12 hours. The mixture was then cooled to room temperature, the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a yellow oily intermediate C8-3 (21 g, 69.7%). 188.1 (M+H) + .

[0819] Step 4: Preparation of intermediate C8

[0820] The synthesis of intermediate C8 referenced intermediate C3, and intermediate C8 was prepared by replacing 1-(tert-butyl)-2-methyl(2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylic acid ester with intermediate C8-3. MS: 160.1 (M+H) + . 1 H NMR (400MHz, CDCl3):d 5.36-5.08(m,1H),3.51-3.29(m,3H),3.25-3.10(m,1H),3.04-2.93(m,1H),2.90-2.7 3(m,1H),2.71-2.59(m,1H),2.28-2.12(m,1H),1.95-1.73(m,4H),1.65-1.53(m,1H).

[0821] Example 1

[0822] Compound I-1: 4'-(4-Acryloylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0823]

[0824] Step 1: Preparation of tert-butyl 4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid

[0825]

[0826] (S)-(1-methylpyrrolidone-2-yl)methanol (105 mg, 0.916 mmol) and 4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A1, 250 mg, 0.458 mmol) were dissolved in anhydrous tetrahydrofuran (20 mL). Cesium carbonate (448 mg, 1.374 mmol) was slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 11 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow oil, 4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (200 mg, 70% yield). MS: 625.1 (M+H) + .

[0827] Step 2: Preparation of tert-butyl piperazine-1-carboxylate

[0828]

[0829] 300 mg (0.480 mmol) of 4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester was dissolved in ethyl acetate (25 mL) and methanol (5 mL). Palladium on carbon (150 mg) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature under hydrogen for 13 hours, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a colorless oily product of 4-(5-amino-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)pyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (286 mg, 100% yield). MS: 595.1 (M+H) + .

[0830] 150 mg (0.252 mmol) of 4-(5-amino-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)pyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester was dissolved in acetic acid (2 mL), and the reaction mixture was stirred at 70 °C for 3 hours. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain a white solid 4-(2'-((((S)-1-methylpyrrolidine-2-yl)methoxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthyl-1,7'-pyrido[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (40 mg, 28% yield). MS: 563.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.22–7.09(m,4H),4.70(t,J=10.0Hz,1H),4.50(dt,J=13.4,7.2Hz,1H),3.85(s,1H),3.77–3 .45(m,8H),3.30(s,3H),3.12–2.92(m,4H),2.82(s,2H),2.37–2.30(m,1H),2.28–1.92(m,4H),1.83(s,3H),1.48(s,9H).

[0831] Step 3: Preparation of 2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-4'-(piperazin-1-yl)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0832]

[0833] 50 mg (0.089 mmol) of tert-butyl piperazine-1-carboxylate was dissolved in dichloromethane (5 mL) and 6N methanol hydrochloride (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and evaporated to dryness, then extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a yellow oil, 2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-4'-(piperazin-1-yl)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (42 mg, 100% yield). MS: 463.1 (M+H) + .

[0834] Step 4: Preparation of 4'-(4-acryloylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0835]

[0836] 2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-4'-(piperazin-1-yl)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (40 mg, 0.086 mmol) and DIEA (33.5 mg, 0.259 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (7.83 mg, 0.086 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. Pre-HPLC analysis of the crude product yielded a white solid, 4'-(4-acryloylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (16 mg, 35.8% yield). MS: 517.1 (M+H) + . 1H NMR (400MHz, Methanol-d4) δ7.17(dd,J=20.2,13.2Hz,4H),6.88–6.68(m,1H),6.26(d,J=16.7Hz,1H),5.79(d,J=10.6Hz,1H),4.61–4.57 (m,3H),3.88(s,3H),3.79–3.47(m,6H),3.34–3.28(m,2H),3.13–2.95(m,4H),2.83(s,2H),2.38(s,1H),2.10–1.96(m,4H),1.84(s,3H).

[0837] Example 2

[0838] Compound I-2: 4'-(4-Acryloylpiperazin-1-yl)-5'-methyl-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0839]

[0840]

[0841] Step 1: Preparation of 4-(5'-methyl-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[0842]

[0843] In 5 mL of 4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (compound I-1 / step 2, 100 mg, 0.178 mmol) DMF, sodium hydrogen (21.32 mg, 0.533 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, and then methyl iodoform (25.2 mg, 0.178 mmol) was added to the reaction mixture, and stirring was continued for 1 hour. After the reaction was complete, the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid, 4-(5'-methyl-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (80 mg, 78% yield). MS: 577.1 (M+H) + .

[0844] Step 2: Preparation of 5'-methyl-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-4'-(piperazin-1-yl)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0845]

[0846] 60 mg (0.104 mmol) of 4-(5'-methyl-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester was dissolved in dichloromethane (5 mL) and 6N methanol hydrochloride (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 5'-methyl-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-4'-(piperazin-1-yl)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (50 mg, 100% yield). MS: 477.1 (M+H) + .

[0847] Step 3: Preparation of 4'-(4-acryloylpiperazin-1-yl)-5'-methyl-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0848]

[0849] 5'-Methyl-2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-4'-(piperazin-1-yl)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (40 mg, 0.084 mmol) and DIEA (33.5 mg, 0.259 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (7.83 mg, 0.086 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid, 4'-(4-acryloylpiperazin-1-yl)-5'-methyl-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (22 mg, 50% yield). MS: 531.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.29–7.07(m,4H),6.81(dd,J=16.8,10.6Hz,1H),6.2 6(dd,J=16.8,1.9Hz,1H),5.80(dd,J=10.6,1.9Hz,1H),4.83–4.67(m,1H),4.55–4. 50(m,1H),3.89(s,3H),3.74–3.69(m,2H),3.65(m,2H),3.51–3.20(m,10H),3.08( s,2H),2.86–2.76(m,2H),2.40–2.37(m,1H),2.31–1.92(m,4H),1.76–1.68(m,3H).

[0850] Example 3

[0851] Compound I-3: 4'-((S)-4-acryloyl-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0852]

[0853]

[0854] Step 1: Preparation of (3S)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester

[0855]

[0856] (S)-(1-methylpyrrolidin-2-yl)methanol (0.339 g, 2.95 mmol) and (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate A2, 1.1 g, 1.964 mmol) were dissolved in anhydrous tetrahydrofuran (20 mL). Cesium carbonate (1.280 g, 3.93 mmol) was slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 11 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (3S)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (800 mg, 63.8% yield). MS: 639.1 (M+H) + .

[0857] Step 2: Preparation of (3S)-3-methyl-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[0858]

[0859] (3S)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (50 mg, 0.078 mmol) and zinc powder (51.2 mg, 0.783 mmol) were added to a mixed solution of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellow solid (3S)-3-methyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (30 mg, 66.5% yield). MS: 577.6 (M+H) + .

[0860] Step 3: Preparation of 4'-((S)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0861]

[0862] (3S)-3-methyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (100 mg, 0.173 mmol) was dissolved in a methanol solution of hydrochloric acid (5 mL). The reaction mixture was stirred at room temperature for 12 hours, and then the reaction mixture solution was concentrated and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 4'-((S)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (83 mg, 100% yield). MS: 477.6 (M+H) + .

[0863] Step 4: Preparation of 4'-((S)-4-acryloyl-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0864]

[0865] 4'-((S)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (100 mg, 0.210 mmol) and DIEA (81 mg, 0.629 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (18.99 mg, 0.210 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC analysis of the crude product yielded a white solid, 4'-((S)-4-acryloyl-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (40 mg, 32.3% yield). MS: 531.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.16–6.97(m,4H),6.92–6.69(m,1H),6.38–6.18(m,1H),5.79(d,J=10.6Hz,1H),4.43–4.17(m,3H),4.02–3.98(m,1H) ,3.90–3.70(m,1H),3.65–3.29(m,4H),3.20–2.92(m,3H),2.78–2.65(m,3 H),2.49(s,3H),2.36–2.29(m,1H),2.16–1.61(m,8H),1.05–0.98(m,3H).

[0866] Compounds I-3a and I-3b

[0867] (R)-4'-((S)-4-acryloyl-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one and (S)-4'-((S)-4-acryloyl-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0868]

[0869] 4'-((S)-4-acryloyl-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (30 mg) was chirally resolved by SFC (column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm column; mobile phase: 40% methanol in carbon dioxide) to give a first component peak of compound I-3a (retention time 1.21 min, 8 mg) and a second component peak of compound I-3b (retention time 1.99 min, 9 mg), both white solids. MS: 531.1 (M+H) + .

[0870] Compound I-3a (retention time 1.21 minutes, 8 mg): 1 H NMR(400MHz, Methanol-d4)δ7.16–6.96(m,4H),6.92–6.69(m,1H),6.38–6.18(m,1H),5.79(d,J=10.6Hz,1H),4.43–4.17(m,3H),4.02–3.96(m,1H) ,3.90–3.70(m,1H),3.65–3.29(m,4H),3.20–2.92(m,3H),2.78–2.64(m,3 H),2.49(s,3H),2.36–2.26(m,1H),2.16–1.61(m,8H),1.05–0.98(m,3H).

[0871] Compound I-3b (retention time 1.99 min, 9 mg): 1H NMR(400MHz, Methanol-d4)δ7.16–6.90(m,4H),6.92–6.69(m,1H),6.38–6.18(m,1H),5.79(d,J=10.6Hz,1H),4.43–4.17(m,3H),4.02–3.91(m,1H) ,3.90–3.75(m,1H),3.65–3.29(m,4H),3.20–2.91(m,3H),2.79–2.66(m,3 H),2.49(s,3H),2.36–2.28(m,1H),2.16–1.61(m,8H),1.05–0.96(m,3H).

[0872] Example 4

[0873] Compound I-4: 4'-((S)-4-(2-fluoroacryloyl)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0874]

[0875] 4'-((S)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (100 mg, 0.210 mmol) and DIEA (81 mg, 0.629 mmol) were dissolved in dichloromethane (5 mL). 2-fluoroacryloyl chloride (intermediate B1, 22.76 mg, 0.210 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. Pre-HPLC analysis of the crude product yielded a white solid, 4'-((S)-4-(2-fluoroacryloyl)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (50 mg, 43.5% yield). MS: 549.1 (M+H) + . 1H NMR(400MHz, Methanol-d4)δ7.14–6.90(m,4H),5.44–5.08(m,2H),4.32–4.26(m,2H),4.08–3.97(m,2H),3.84–3.31(m,5H),3 .18–2.92(m,2H),2.80–2.71(m,3H),2.50(s,3H),2.37–2.29(m,1H),2.06–1.89(m,2H),1.77–1.69(m,6H),1.29–1.19(m,4H).

[0876] Compounds I-4a and I-4b

[0877] (R)-4'-((S)-4-(2-fluoroacryloyl)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one and (S)-4'-((S)-4-(2-fluoroacryloyl)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0878]

[0879] 4'-((S)-4-(2-fluoroacryloyl)-2-methylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-6'-one (40 mg) was chirally resolved by SFC (column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm column, mobile phase: 40% methanol in carbon dioxide) to give a first component peak of compound I-4a (retention time 0.93 min, 12 mg) and a second component peak of compound I-4b (retention time 1.43 min, 10 mg), both white solids. MS: 549.1 (M+H) + .

[0880] Compound I-4a (retention time 0.93 min, 12 mg): 1H NMR (400MHz, Methanol-d4) δ7.14–6.90(m4H),5.44–5.08(m,2H),4.32–4.26(m,2H),4.08–3.97(m,2H),3.84–3.31(m,5H),3. 18–2.92(m,2H),2.80–2.71(m,3H),2.50(s,3H),2.37–2.29(m,1H),2.06–1.89(m,2H),1.77–1.69(m,6H),1.29–1.19(m,4H).

[0881] Compound I-4b (retention time 1.43 min, 10 mg): 1 H NMR(400MHz, Methanol-d4)δ7.14–6.90(m,4H),5.44–5.08(m,2H),4.32–4.26(m,2H),4.08–3.97(m,2H),3.84–3.31(m,5H),3 .18–2.92(m,2H),2.80–2.71(m,3H),2.50(s,3H),2.37–2.29(m,1H),2.06–1.89(m,2H),1.77–1.69(m,6H),1.29–1.19(m,4H).

[0882] Example 5

[0883] Compound I-5: 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0884]

[0885] Step 1: Preparation of (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[0886]

[0887] (S)-(1-methylpyrrolidin-2-yl)methanol (413 mg, 3.59 mmol) and (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 700 mg, 1.196 mmol) were dissolved in anhydrous tetrahydrofuran (2 mL). Cesium carbonate (780 mg, 2.393 mmol) was slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 11 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (700 mg, 88% yield). MS: 664.1 (M+H) + .

[0888] Step 2: Preparation of (2S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[0889]

[0890] (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (700 mg, 1.055 mmol) and zinc powder (207 mg, 3.16 mmol) were added to a mixture of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellow solid (2S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (500 mg, 78.7% yield). MS: 602.3 (M+H) + .

[0891] Step 3: Preparation of 2-((2S)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0892]

[0893] (2S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (350 mg, 0.582 mmol) was dissolved in dichloromethane (5 mL) and 6N methanol hydrochloride (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 2-((2S)-4-(2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (292 mg, 100% yield). MS: 502.3 (M+H) + .

[0894] Step 4: Preparation of 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0895]

[0896] 2-((2S)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (100 mg, 0.199 mmol) and TEA (60.5 mg, 0.598 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (18.04 mg, 0.199 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (55 mg, 50% yield). MS: 556.1 (M+H). + . 1 H NMR (400MHz, Methanol-d4) δ7.17–6.99(m,4H),6.79(s,1H),6.27(d,J=16.7Hz,1H),5.81(d,J=10.6Hz,1H),4.39–4.22(m,2 H),4.20–3.68(m,3H),3.59–3.32(m,3H),3.15–2.67(m,8H),2.49(s,3H),2.37–2.34(m,1H),2.15–1.60(m,8H),1.29(s,1H).

[0897] Compound I-5a and Compound I-5b

[0898] 2-((S)-1-Acryloyl-4-((R)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-Acryloyl-4-((S)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0899]

[0900]

[0901] 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (50 mg) was chirally resolved by SFC (column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm column, mobile phase: 40% methanol in carbon dioxide) to give a first component peak of compound I-5a (retention time 1.24 min, 11 mg) and a second component peak of compound I-5b (retention time 2.56 min, 8 mg), both white solids. MS: 556.1 (M+H) + .

[0902] Compound I-5a (retention time 1.24 min, 11 mg). 1 H NMR (400MHz, Methanol-d4) δ7.17–6.99(m,4H),6.79(s,1H),6.27(d,J=16.7Hz,1H),5.81(d,J=10.6Hz,1H),4.39–4.22(m,2 H),4.20–3.68(m,3H),3.59–3.32(m,3H),3.15–2.67(m,8H),2.49(s,3H),2.37–2.34(m,1H),2.15–1.60(m,8H),1.29(s,1H).

[0903] Compound I-5b (retention time 2.56 min, 8 mg). 1 H NMR (400MHz, Methanol-d4) δ7.17–6.99(m,4H),6.79(s,1H),6.27(d,J=16.7Hz,1H),5.81(d,J=10.6Hz,1H),4.39–4.22(m,2 H),4.20–3.68(m,3H),3.59–3.32(m,3H),3.15–2.67(m,8H),2.49(s,3H),2.37–2.34(m,1H),2.15–1.60(m,8H),1.29(s,1H).

[0904] Example 6

[0905] Compound I-6(481): 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0906]

[0907] 2-((2S)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (100 mg, 0.199 mmol) and TEA (60.5 mg, 0.598 mmol) were dissolved in dichloromethane (5 mL). 2-fluoroacryloyl chloride (intermediate B1, 21.63 mg, 0.199 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (55 mg, 50% yield). MS: 574.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.26–7.06(m,4H),5.30–5.20(m,2H),4.48–4.27(m,2H),3.93–3.69(m,2H),3.52 –3.41(m,1H),3.37–3.21(m,1H),3.17–2.67(m,9H),2.49(s,3H),2.35(m,1H),2.18–1.58(m,8H),1.29(s,2H).

[0908] Compound I-6a and Compound I-6b

[0909] 2-((S)-1-(2-fluoroacryloyl)-4-((R)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-(2-fluoroacryloyl)-4-((S)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0910]

[0911] 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (50 mg) was chirally resolved by SFC (column: Dr. Maish Reprosil Chiral-JM, 250x25 mm, 10 μm column, mobile phase: 40% methanol in carbon dioxide) to give the first component peak, compound I-6a (retention time 1.01 min, 13 mg), and the second component peak, compound I-6b (retention time 2.17 min, 9 mg), both white solids. MS: 574.1 (M+H) + .

[0912] Compound I-6a (retention time 1.01 min, 13 mg). 1 H NMR(400MHz, Methanol-d4)δ7.26–7.06(m,4H),5.30–5.20(m,2H),4.48–4.27(m,2H),3.93–3.69(m,2H),3.52 –3.41(m,1H),3.37–3.21(m,1H),3.17–2.67(m,9H),2.49(s,3H),2.35(m,1H),2.18–1.58(m,8H),1.29(s,2H).

[0913] Compound I-6b (retention time 2.17 min, 9 mg). 1H NMR(400MHz, Methanol-d4)δ7.26–7.06(m,4H),5.30–5.20(m,2H),4.48–4.27(m,2H),3.93–3.69(m,2H),3.52 –3.41(m,1H),3.37–3.21(m,1H),3.17–2.67(m,9H),2.49(s,3H),2.35(m,1H),2.18–1.58(m,8H),1.29(s,2H).

[0914] Example 7

[0915] Compound I-7: 4'-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0916]

[0917] The synthesis of compound I-7 is based on compound I-5, and compound I-7 was prepared by replacing (2R,5S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylic acid tert-butyl ester (intermediate A5) with (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0918] Compound I-7 (11.6 mg). MS: 545.1 (M+H) + . 1H NMR(400MHz,DMSO-d6)δ7.29–7.06(m,4H),6.80(s,1H),6.15–6.98(m,1H),5.71–5.65(m,1H),4 .78(s,1H),4.52–4.49(m,1H),4.46–4.28(m,2H),4.00–3.87(m,1H),3.76–3.65(m,2H),3.52-3 .41(m,3H),3.21–3.02(m,2H),2.90–2.85(m,2H),2.81(s,1H),2.73(s,1H),2.41(s,3H),2.19( s,1H),2.02(s,1H),1.93–1.77(m,3H),1.74–1.67(m,1H),1.18–1.09(m,3H),1.00–0.89(m,3H).

[0919] Example 8

[0920] Compound I-8: 4'-(2-Acryloyl-2,7-diazaspiro[3.5]non-7-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0921]

[0922] The synthesis of compound I-8 is based on compound I-5, and compound I-8 was prepared by replacing (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (intermediate A6) with (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0923] Compound I-8 (10.6 mg). MS: 557.1 (M+H) + . 1H NMR(400MHz, DMSO-d6)7.13–7.02(m,4H),6.32(dd,J=17.0,10.3Hz,1H),6.10 (d,J=17.0Hz,1H),5.66–5.59(m,1H),4.58–4.36(m,2H),4.09–3.83(m,10H), 3.68(s,1H),3.57(s,1H),3.43–3.38(m,3H),3.25(s,2H),3.15–3.05(m,1H), 2.92–2.71(m,5H),2.21–2.16(m,1H),1.94(s,1H),1.86(s,1H),1.71(s,4H).

[0924] Example 9

[0925] Compound I-9: 2-((2S)-1-(but-2-acyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0926]

[0927] 2-((2S)-4-(2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (50 mg, 0.100 mmol) and TEA (60.5 mg, 0.598 mmol) were dissolved in dichloromethane (5 mL). Butyryl-2-acetylacetyl chloride (intermediate B2, 10.63 mg, 0.100 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to obtain a crude product. Pre-HPLC analysis of the crude product yielded a white solid, 2-((2S)-1-(but-2-ynyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (5 mg, 10% yield). MS: 568.1 (M+H) + . 1H NMR(400MHz, Methanol-d4)δ7.17–6.98(m,4H),4.64–4.56(m,3H),4.41–4.30(m,3H),4.19–3.70(m, 2H),3.60–3.33(m,1H),3.17–2.74(m,7H),2.54–2.45(m,4H),2.32–1.60(m,9H),1.30–1.15(m,3H).

[0928] Example 10

[0929] Compounds I-10a and I-10b: 2-((S)-1-acryloyl-4-((S)-2'-methoxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-acryloyl-4-((R)-2'-methoxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0930]

[0931] The synthesis of compounds I-10a and I-10b is based on compound I-5, and compounds I-10a and I-10b are prepared by using (2S)-2-(cyanomethyl)-4-(2-methoxy-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A11) instead of (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (compound I-5 / step 1).

[0932] Compound I-10a (retention time 4.74 min, 7.6 mg). MS: 473.1 (M+H) + . 1H NMR (400MHz, Methanol-d4) δ7.15–6.97 (m, 4H), 6.80 (s, 1H), 6.28–6.01 (m, 1H), 5.83–5.71 (m, 1H), 5.24–4.86 (m, 1H) ), 4.68–4.30(m,1H),4.03–3.88(m,4H),3.66–3.28(m,6H),3.13–2.88(m,2H),2.83–2.74(m,2H),2.20–1.72(m,4H).

[0933] Compound I-10b (retention time 5.44 min, 11.6 mg). MS: 473.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.15–6.97(m,4H),6.80(s,1H),6.28–6.01(m,1H),5.83–5.71(m,1H),5.24–4.86(m,1H ),4.68–4.30(m,1H),4.03–3.88(m,4H),3.66–3.28(m,6H),3.13–2.88(m,2H),2.83–2.74(m,2H),2.20–1.72(m,4H).

[0934] Example 11

[0935] Compound I-11: 2-((2S)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-((E)-4,4,4-trifluoro-2-enyl)piperazin-2-yl)acetonitrile

[0936]

[0937] The synthesis of compound I-11 is based on compound I-6, and compound I-11 was prepared by using (E)-4,4,4-trifluoro-2-enoyl chloride (intermediate B3) instead of 2-fluoroacryloyl chloride (intermediate B1).

[0938] Compound I-11 (25.4 mg). MS: 625.1 (M+H) + . 1H NMR (400MHz, DMSO-d6) δ7.54–7.35(m,1H),7.27–7.02(m,4H),6.84(dd,J=15.6 ,7.6Hz,1H),5.05(s,1H),4.80(s,1H),4.58–4.48(m,1H),4.43(s,1H),3.75(s, 2H),3.59–3.41(m,4H),3.36–3.20(m,1H),3.13(s,2H),2.99–2.95(m,1H),2.9 2–2.85(m,5H),2.76–2.53(m,2H),2.23(s,1H),2.04(s,2H),1.74–1.54(m,6H).

[0939] Example 12

[0940] Compound I-12: 4'-((R)-4-acryloyl-3-(fluoromethyl)piperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-6'-one

[0941]

[0942] The synthesis of compound I-12 is based on compound I-5, and compound I-12 was prepared by replacing (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(fluoromethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A3) with (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0943] Compound I-12 (10.6 mg). MS: 549.1 (M+H) + . 1H NMR (400MHz, DMSO-d6) δ7.27–7.04 (m, 4H) 6.81 (dd, J=16.4, 10.6Hz, 1H), 6.15 (d, J= 16.2Hz,1H),5.72–5.54(m,1H),4.92(s,1H),4.63(s,2H),4.40–4.13(m,2H),4.08– 3.98(m,1H),3.89–3.66(m,2H),3.58–3.45(m,1H),3.16–3.08(m,2H),2.97–2.92(m ,1H),2.89–2.61(m,4H),2.55(s,1H),2.34(s,3H),2.18(s,1H),2.01–1.48(m,8H).

[0944] Example 13

[0945] Compound I-13: 4'-((3R,5S)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0946]

[0947] The synthesis of compound I-13 is based on compound I-5, and compound I-13 was prepared by replacing (2R,6S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (intermediate A4) with (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0948] Compound I-13 (9.1 mg). MS: 545.1 (M+H) + . 1H NMR (400MHz, DMSO-d6) δ7.18–7.06(m,4H),6.77(dd,J=16.5,10.5Hz,1H),6.16(d,J=16.6Hz,1H),5.73–5.68(m,1H),4. 48–4.35(m,4H),4.19–3.99(m,1H),3.86(s,1H),3.42–3.26(m,2H),3.18–3.09(m,2H),2.91–2.85(m,3H),2.83–2.78(m 1H),2.74(s,3H),2.42(s,1H),2.23(s,1H),2.03(s,1H),1.89(s,3H),1.71(s,3H),1.32–1.25(m,3H),1.14–1.10(m,3H).

[0949] Example 14

[0950] Compound I-14: 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0951]

[0952]

[0953] Step 1: Preparation of (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-2-(((S)-1-methylpyrrolidone-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester

[0954]

[0955] (S)-(1-methylpyrrolidin-2-yl)methanol (114 mg, 0.992 mmol) and (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A12, 200 mg, 0.331 mmol) were dissolved in anhydrous tetrahydrofuran (5 mL). Cesium carbonate (215 mg, 0.661 mmol) was slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 11 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-2,3-dihydro-1H-inden-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (113 mg, 50% yield). MS: 684.1 (M+H) + .

[0956] Step 2: Preparation of (2S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic [indene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid benzyl ester

[0957]

[0958] ((2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-2,3-dihydro-1H-indene-1-yl)methyl)-2-(((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (226 mg, 0.331 mmol) and zinc powder (103 mg, 1.18 mmol) were added to a mixed solution of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid benzyl ester (120 mg, 58.4% yield). MS: 622.1 (M+H) + .

[0959] Step 3: Preparation of 2-((2S)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0960]

[0961] (2S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid benzyl ester (210 mg, 0.338 mmol) was dissolved in ethanol (5 mL), and palladium on carbon (711 mg) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature under hydrogen atmosphere for 2 hours, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a yellow oily substance, 2-((2S)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (160 mg, 97% yield). MS: 488.1 (M+H) + .

[0962] Step 4: Preparation of 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0963]

[0964] 2-((2S)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (90 mg, 0.185 mmol) and TEA (56.0 mg, 0.554 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (16.7 mg, 0.185 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (50 mg, 50% yield). MS: 542.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.42–6.65(m,5H),6.28–6.15(m,1H),5.82–5.73(m,1H),5.06(s,1H),4.71–4.64(m,1H),4.52 (s,1H),4.04–3.95(m,3H),3.78–3.67(m,2H),3.37–3.35(m,1H),3.34–2.69(m,13H),2.43–2.31(m,1H),2.11–1.96(m,5H).

[0965] Example 15

[0966] Compound I-15: 2-((2S)-4-acryloyl-1-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0967]

[0968] The synthesis of compound I-15 is based on compound I-5, which was prepared by replacing (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3) with (3S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0969] Compound I-15 (2.1 mg). MS: 556.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.13–7.02(m,4H),6.81–6.71(m,1H),6.45–6.07(m,1H),5.82–5.45(m ,1H),4.57–4.21m,6H),3.89–3.32(m,6H),3.09–2.58(m,5H),2.34–1.67(m,8H),1.30–1.21(m,4H).

[0970] Example 16

[0971] Compound I-16: 4'-((1R,5S)-8-acryloyl-3,8-diazabicyclo[3.2.1]octane-3-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0972]

[0973] The synthesis of compound I-16 is based on compound I-5, and compound I-16 was prepared by using (1R, 5S)-3-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (intermediate A7) instead of (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0974] Compound I-16 (11.6 mg). MS: 543.1 (M+H) + . 1H NMR(400MHz, Methanol-d4)δ7.14–6.95(m,4H),6.75(dd,J=16.8,10.4Hz,1H),6.34(d,J=16.7 Hz,1H),5.80–5.45(m,1H),4.77–4.68(m,1H),4.61(s,1H),4.45–4.38(m,1H),4.32–4.13(m,2 H),3.72–3.69(m,1H),3.66–3.50(m,1H),3.44–3.34(m,1H),3.23–3.05(m,2H),2.94–2.87(m, 1H),2.81–2.61(m,3H),2.51(s,3H),2.38–2.34(m,1H),2.32–2.15(m,2H),2.13–1.63(m,8H).

[0975] Example 17

[0976] Compound I-17: 4'-((1R,5S)-3-acryloyl-3,8-diazabicyclo[3.2.1]octane-8-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0977]

[0978] The synthesis of compound I-17 is based on compound I-5, and compound I-17 was prepared by using (1R, 5S)-8-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester (intermediate A8) instead of (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[0979] Compound I-17 (15.7 mg). MS: 543.1 (M+H) + . 1H NMR(400MHz, Methanol-d4)δ7.15–6.98(m,4H),6.81–6.63(m,1H),6.23–6.10(m,1H),5.76–5.65(m,1H),4.69(s, 1H),4.55–4.49(m,1H),4.32–4.15(m,3H),4.02–3.42(m,3H),3.35–3.19(m,2H),3.11–2.99(m,1H),2.93–2.87(m 1H),2.80–2.75(m,3H),2.51(s,3H),2.39–2.34(m,1H),2.29–1.61(m,9H).

[0980] Example 18

[0981] Compound I-18: 2-((2S)-1-((E)-4-methoxy-2-enyl)-4-(2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[0982]

[0983] The synthesis of compound I-18 is based on compound I-6, and compound I-18 was prepared by replacing 2-fluoroacryloyl chloride (intermediate B1) with (E)-4-methoxy-2-enoyl chloride (intermediate B4).

[0984] Compound I-18 (20.4 mg). MS: 600.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ7.30–7.00(m,4H),6.67–6.45(m,2H),5.06(s,1H),4.62–4.24(m,4H),4.08(s,2H),3.72–3.59(m,6H),3.37(s,1H), 3.30–3.27(m,3H),3.10–3.05(m,3H),2.97–2.94(m,1H),2.91(s,3H), 2.75–2.69(m,3H),2.22(s,1H),2.04–1.95(m,2H),1.79–1.68(m,5H).

[0985] Example 19

[0986] Compound I-19: 4'-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[0987]

[0988] Step 1: Preparation of (R)-4-acryloyl-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester

[0989]

[0990] (R)-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (2.0 g, 9.26 mmol) and TEA (1.87 g, 18.52 mmol) were dissolved in dichloromethane (25 mL). Acryloyl chloride (922 mg, 10.19 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product, which was purified by silica gel column chromatography to give a colorless oil (R)-4-acryloyl-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (1.8 g, 72% yield). MS: 271.3 (M+H) + .

[0991] Step 2: Preparation of (R)-1-(3-(hydroxymethyl)piperazin-1-yl)prop-2-en-1-one

[0992]

[0993] (R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-carboxylic acid tert-butyl ester (1.8 g, 6.67 mmol) was dissolved in dichloromethane (15 mL) and trifluoroacetic acid (10 mL). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated to dryness and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and dried over sodium sulfate, then filtered and concentrated to give a yellow oil (R)-1-(3-(hydroxymethyl)piperazin-1-yl)prop-2-en-1-one (1.7 g, 100% yield). MS: 171.3 (M+H) + .

[0994] Step 3: Preparation of methyl 1-((6-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2-chloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0995]

[0996] (R)-1-(3-(hydroxymethyl)piperazin-1-yl)prop-2-en-1-one (120 mg, 0.51 mmol) and methyl 1-((2,6-dichloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (intermediate A3-5, 200 mg, 0.51 mmol) were dissolved in anhydrous dichloromethane (5 mL). TEA (78 mg, 0.76 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a colorless oily methyl 1-((6-(((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2-chloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (150 mg, 42% yield). MS: 530.2 (M+H) + .

[0997] Step 4: Preparation of methyl 1-((6-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid

[0998]

[0999] (S)-(1-methylpyrrolidone-2-yl)methanol (54 mg, 0.47 mmol) and methyl 1-((6-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2-chloro-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (150 mg, 0.28 mmol) were dissolved in anhydrous tetrahydrofuran (10 mL). Cesium carbonate (153 mg, 0.47 mmol) was slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 11 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a colorless oil, methyl 1-((6-(((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid (120 mg, 70% yield). MS: 609.3 (M+H) + .

[1000] Step 5: Preparation of 4'-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[1001]

[1002] 1-((6-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2-((S)-1-methylpyrrolidine-2-yl)methoxy)-5-nitropyrimidin-4-yl)methyl)-1,2,3,4-tetrahydronaphthalene-1-carboxylic acid methyl ester (120 mg, 0.20 mmol) and zinc powder (30 mg, 0.46 mmol) were added to a mixed solution of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 50°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellowish-white solid 4'-((R)-4-acryloyl-2-(hydroxymethyl)piperazin-1-yl)-2'-(((S)-1-methylpyrrolidine-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one (20 mg, 18.7% yield). MS: 547.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.14–6.95(m,4H),6.81(dd,J=16.7,10.5Hz,1H),6.30(d,J=16.7Hz,1 H),5.94–5.79(m,1H),4.79–4.50(m,6H),4.05–3.45(m,8H),3.28–2.65(m,9H),2.49–1.78(m,6H).

[1003] Example 20

[1004] Compounds I-20a and I-20b: 2-((S)-1-acryloyl-4-((R)-2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-acryloyl-4-((S)-2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1005]

[1006]

[1007] Step 1: Preparation of (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[1008]

[1009] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 200 mg, 0.342 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL). 2-methyl-1,2,3,4-tetrahydroisoquinoline-5-ol (55.8 mg, 0.342 mmol) and sodium hydroxide (13.67 mg, 0.342 mmol) were slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 4 hours. The reaction was then quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (197 mg, 91% yield). MS: 712.1 (M+H) + .

[1010] Step 2: Preparation of (2S)-2-(cyanomethyl)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[1011]

[1012] (2S)-2-(cyanomethyl)-4-(6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-2-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (190 mg, 0.267 mmol) and zinc powder (87 mg, 1.335 mmol) were added to a mixed solution of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellow solid (2S)-2-(cyanomethyl)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (200 mg, 115% yield). MS: 550.1 (M+H) + .

[1013] Step 3: Preparation of 2-((2S)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1014]

[1015] (2S)-2-(cyanomethyl)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.308 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 2-((2S)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (175 mg, 103% yield). MS: 550.1 (M+H) + .

[1016] Step 4: Preparation of 2-((S)-1-acryloyl-4-((R)-2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-acryloyl-4-((S)-2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1017]

[1018] 2-((2S)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (175 mg, 0.318 mmol) and DIEA (82 mg, 0.637 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (43.2 mg, 0.478 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. Pre-HPLC was used to prepare compounds I-20a (retention time 4.08 min, 3 mg) and I-20b (retention time 4.22 min, 4 mg), both white solids. MS: 604.1 (M+H) + .

[1019] Compound I-20a (retention time 4.08 min, 3 mg). 1 H NMR(400MHz,DMSO-d6)δ7.12–6.98(m,5H),6.92–6.89(m,2H),6.81(s,1H),6. 15–6.09(m,1H),5.73–5.61(m,1H),4.93(s,1H),4.65(s,1H),4.30(s,1H),3.6 5(s,2H),3.54-3.51(m,3H),3.41-3.25(m,2H),3.15(s,1H),2.90–2.79(m,3H) ,2.75–2.68(m,2H),2.32(s,3H),2.04–1.95(m,2H),1.76(s,3H),1.22(s,2H).

[1020] Compound I-20b (retention time 4.22 min, 4 mg). 1H NMR(400MHz,DMSO-d6)δ7.12–6.98(m,5H),6.92–6.89(m,2H),6.81(s,1H),6. 15–6.09(m,1H),5.73–5.61(m,1H),4.93(s,1H),4.65(s,1H),4.30(s,1H),3.6 5(s,2H),3.54-3.51(m,3H),3.41-3.25(m,2H),3.15(s,1H),2.90–2.79(m,3H) ,2.75–2.68(m,2H),2.32(s,3H),2.04–1.95(m,2H),1.76(s,3H),1.22(s,2H).

[1021] Example 21

[1022] Compound I-21: 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1023]

[1024] The synthesis of compound I-21 is based on compound I-14, and compound I-21 is prepared by using 2-fluoroacryloyl chloride (intermediate B1) instead of acryloyl chloride.

[1025] Compound I-21 (46.5 mg). MS: 560.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.34–6.88(m,4H)5.32–5.18(m,2H),4.73–4.61(m,1H), 4.53(s,1H),4.08–3.99(m,3H),3.79–3.64(m,2H),3.49–2.90(m,13H),2.43–2.21(m 1H),2.13–2.09(m,6H).

[1026] Example 22

[1027] Compound I-22: 2-((2S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxo-5',8'-dihydro-6'H-spirocyclic[chroman-4,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1028]

[1029] The synthesis of compound I-22 is based on compound I-14. Compound I-22 was prepared by replacing (2S)-4-(2-chloro-6-((4-(methoxycarbonyl)chroman-4-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A13) with (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-indene-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A12).

[1030] Compound I-22 (4.6 mg). MS: 558.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.20–7.10(m,2H),7.01–6.69(m,2H),6.29–6.11(m,1H),5.83–5.76(m,1H),5.09(s, 1H),4.58(s,3H),4.41–3.81(m,6H),3.64–3.51(m,1H),3.48–2.76(m,8H),2.49–1.66(m,6H),1.31–1.20(m,3H).

[1031] Example 23

[1032] Compound I-23: 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-((2-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1033]

[1034] The synthesis of compound I-23 is based on compound I-20, and compound I-23 was prepared by using 2-fluoroacryloyl chloride (intermediate B1) instead of acryloyl chloride.

[1035] Compound I-23 (14.83 mg). MS: 622.1 (M+H) + . 1H NMR(400MHz, DMSO-d6)δ7.39–7.19(m,2H),7.11(dq,J=14.1,8.2,7.6Hz,5H),5.44–5.11(m,2H),4. 54(d,J=14.2Hz,1H),4.45–4.25(m,1H),4.02–3.43(m,6H),3.31–2.53(m,14H),2.14–1.57(m,4H).

[1036] Example 24

[1037] Compound I-24: 4'-((R)-4-acryloyl-3-(hydroxymethyl)piperazin-1-yl)-2'-(((S)-1-methylpyrrolidone-2-yl)methoxy)-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-6'-one

[1038]

[1039] The synthesis of compound I-24 is based on compound I-5, and compound I-24 was prepared by replacing (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3) with (2R)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3).

[1040] Compound I-24 (10.67 mg). MS: 547.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.15–6.96(m,4H),6.80(s,1H),6.25–6.01(m,1H),5.94–5.59(m,1H),4.82–4. 41(m,3H),4.40–3.97(m,2H),3.95–3.36(m,3H),3.33–2.67(m,10H),2.51–1.77(m,9H),1.31–1.20(m,2H).

[1041] Example 25

[1042] Compound I-25: 2-((2S)-1-acetyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-2,3,5',8'-tetrahydro-6'H-spirocyclic[inden-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1043]

[1044] The synthesis of compound I-25 is based on compound I-14, and compound I-25 is prepared by using acetyl chloride instead of acryloyl chloride.

[1045] Compound I-25 (20 mg). MS: 530.1 (M+H) + . 1 H NMR (400MHz, Methanol-d4) δ7.34–6.86(m,4H),5.00–4.86(m,1H),4.61–4.43(m,4H),4.21–3.57(m,5H),3.43–2.84(m,11H),2.55–1.93(m,9H).

[1046] Example 26

[1047] Compound I-26: 2-((2S)-4-(2'-(((2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile

[1048]

[1049] The synthesis of compound I-26 references compound I-14, by using (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) instead of ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro) Compound I-26 was prepared by replacing (S)-(1-methylpyrrolidone-2-yl)methanol with (2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methanol (intermediate C1) and acryloyl chloride with 2-fluoroacryloyl chloride (intermediate B1).

[1050] Compound I-26 (4.6 mg). MS: 592.1 (M+H) + . 1 H NMR(400MHz,DMSO-d6)δ7.29–7.02(m,4H),5.42–5.03(m,3H),4.26–4.19(m,1H),4.17 –4.02(m,1H),3.81–3.47(m,2H),3.34–2.54(m,17H),2.31(s,4H),2.13–1.57(m,2H).

[1051] Example 27

[1052] Compound I-27: 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-(((2S,4R)-4-methoxy-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1053]

[1054] The synthesis of compound I-27 references compound I-14, by using (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) instead of ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro- Compound I-27 was prepared by replacing (S)-(1-methylpyrrolidine-2-yl)methanol with (2S,4R)-4-methoxy-1-methylpyrrolidine-2-yl)methanol (intermediate C2) and acryloyl chloride with 2-fluoroacryloyl chloride (intermediate B1).

[1055] Compound I-27 (14.6 mg). MS: 604.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ7.31–7.04(m,4H),5.45–5.13(m,2H),4.64–4.38(m,2H),4.09(s, 1H),3.89(s,1H),3.82–3.61(m,2H),3.59–2.65(m,19H),2.32(s,1H),2.07–1.59(m,5H).

[1056] Example 28

[1057] Compound I-28: 2-((2S)-1-acryloyl-4-(2'-(((2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1058]

[1059] The synthesis of compound I-28 is based on compound I-14. Compound I-28 was prepared by replacing ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) with ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-indene-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A12) and ((2S,4S)-4-fluoro-1-methylpyrrolidine-2-yl)methanol (intermediate C1) with (S)-(1-methylpyrrolidine-2-yl)methanol.

[1060] Compound I-28 (14.6 mg). MS: 574.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ7.32–7.03(m,4H),6.84(s,1H),6.18(d,J=16.7Hz,1H),5.77(d,J=10.3Hz,1H),5. 46(m,1H),5.16–4.68(m,1H),4.65–4.32(m,2H),4.20–3.44(m,9H),3.33–2.61(m,11H),2.21–1.60(m,5H).

[1061] Example 29

[1062] Compound I-29: 2-((2S)-1-acryloyl-4-(2'-(((2S,4R)-4-methoxy-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1063]

[1064] The synthesis of compound I-29 is based on compound I-14. Compound I-29 was prepared by replacing ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) with ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-indene-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A12) and ((2S,4R)-4-methoxy-1-methylpyrrolidine-2-yl)methanol (intermediate C2) with (S)-(1-methylpyrrolidine-2-yl)methanol.

[1065] Compound I-29 (11.6 mg). MS: 586.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ7.33–7.05(m,4H),6.84(s,1H),6.18(d,J=16.7Hz,1H),5.77(d,J=10.4Hz,1 H),4.94–4.81(mz,1H),4.67–4.30(m,2H),4.16–3.42(m,9H),3.32–2.60(m,13H),2.39–1.59(m,6H).

[1066] Example 30

[1067] Compound I-30: 2-((2S)-4-(2'-(((2S,4R)-4-methoxy-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-((E)-4-methoxy-2-enyl)piperazin-2-yl)acetonitrile

[1068]

[1069] The synthesis of compound I-30 references compound I-14, by using (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) instead of ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-indene) Compound I-30 was prepared by replacing (S)-(1-methylpyrrolidone-2-yl)methanol with (2S,4R)-4-methoxy-1-methylpyrrolidone-2-yl)methanol (intermediate C2) and (E)-4-methoxy-2-enoyl chloride (intermediate B4) with acryloyl chloride.

[1070] Compound I-30 (8.4 mg). MS: 630.1 (M+H) + . 1 H NMR(400MHz,DMSO-d6)δ7.26–7.04(m,4H),6.66–6.21(m,1H),5.06(s,1H),4.43–3.93(m,3H),3.89–3.43(m,9H), 3.35–3.19(m,6H),3.16–3.14(m,3H),3.12–2.52(m,5H),2.29–2.21(m,3H),2.19–1.92(m,2H),1.92–1.64(m,5H).

[1071] Example 31

[1072] Compound I-31: 2-((2S)-1-acryloyl-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1073]

[1074]

[1075] Step 1: Preparation of (2S)-2-(cyanomethyl)-4-(2-((2-ethylpyridin-3-yl)oxy)-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester

[1076]

[1077] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14, 600 mg, 0.969 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL). 2-ethylpyridin-3-ol (239 mg, 1.938 mmol) and sodium hydroxide (116 mg, 2.91 mmol) were slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 4 hours, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(2-((2-ethylpyridin-3-yl)oxy)-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (580 mg, 85% yield). MS: 706.1 (M+H) + .

[1078] Step 2: Preparation of (2S)-2-(cyanomethyl)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid benzyl ester

[1079]

[1080] (2S)-2-(cyanomethyl)-4-(2-((2-ethylpyridin-3-yl)oxy)-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (580 mg, 0.822 mmol) and zinc powder (161 mg, 2.46 mmol) were added to a mixture of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid benzyl ester (280 mg, 52.9% yield). MS: 644.1 (M+H) + .

[1081] Step 3: Preparation of 2-((2S)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1082]

[1083] (2S)-2-(cyanomethyl)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid benzyl ester (280 mg, 0.435 mmol) was dissolved in ethanol (15 mL), and palladium on carbon (231 mg) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature under hydrogen atmosphere for 12 hours, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a yellow oily substance, 2-((2S)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (222 mg, 100% yield). MS: 510.1 (M+H) + .

[1084] Step 4: Preparation of 2-((2S)-1-acryloyl-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1085]

[1086] 2-((2S)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (100 mg, 0.196 mmol) and TEA (19.86 mg, 0.196 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (17.76 mg, 0.196 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. Pre-HPLC analysis of the crude product yielded a white solid, 2-((2S)-1-acryloyl-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (14 mg, 7.3% yield). MS: 564.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ8.36–8.15(m,1H),7.61–7.57(m,1H),7.36–7.21(m,1H),7.17–7.89(m,5H),6.76(s,1H),6.25–6 .10(m,1H),5.80–5.54(m,1H),4.69(s,4H),3.69(m,3H),3.24–2.94(m,2H),2.78(m,5H),2.15–1.78(m4H),1.42–1.15(m,4H).

[1087] Example 32

[1088] Compound I-32: 2-((2S)-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile

[1089]

[1090] The synthesis of compound I-32 is based on compound I-31, and compound I-32 is prepared by using 2-fluoroacryloyl chloride (intermediate B1) instead of acryloyl chloride.

[1091] Compound I-32 (9.6 mg). MS: 582.1 (M+H) +. 1 H NMR(400MHz, Methanol-d4)δ8.52(d,J=5.0Hz,1H),8.24–8.16(m,1H),7.75(dd,J=8.4,5.7Hz,1H),7.26–7.07(m,4H),5.39–5.16(m,2H),4.82–4 .58(m,3H),4.21–3.90(m,2H),3.78–3.61(m,1H),3.54–3.39(m,1H),3.2 3–3.18(m,1H),3.10–2.58(m,7H),2.15–1.76(m,4H),1.44–1.24(m,3H).

[1092] Example 33

[1093] Compound I-33: 2-((2S)-1-acetyl-4-(2'-((2-ethylpyridin-3-yl)oxy)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1094]

[1095] The synthesis of compound I-33 is based on compound I-31, and compound I-33 was prepared by using acetyl chloride instead of acryloyl chloride.

[1096] Compound I-33 (9.6 mg). MS: 552.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ8.52(d,J=5.6Hz,1H),8.27–8.10(m,1H),7.85–7.67(m,1H),7.28–7.04(m,4H),4. 62–4.31(m,1H),3.88–3.79(m,2H),3.76–3.33(m,3H),3.25–2.47(m,9H),2.33–1.76(m,6H),1.31–1.21(m,4H).

[1097] Example 34

[1098] Compound I-34: 2-((2S)-1-(2-fluoroacryloyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-6'-oxy-5',8'-dihydro-6'H-spirocyclic [chroman-4,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1099]

[1100] The synthesis of compound I-34 is based on compound I-22, and compound I-34 was prepared by using 2-fluoroacryloyl chloride (intermediate B1) instead of acryloyl chloride.

[1101] Compound I-34 (14.6 mg). MS: 576.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.28–7.11(m,2H),6.97–6.74(m,2H),5.32–5.09(m,2H),4.82 –4.42(m,3H),4.36–3.55(m,9H),3.51–3.22(m,3H),3.15–2.81(m,5H),2.45–1.86(m,7H).

[1102] Example 35

[1103] Compound I-35: 2-((2S)-1-((E)-4-(dimethylamino)but-2-enyl)-4-(2'-((2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1104]

[1105] 2-((2S)-4-(2'-(((2S,4S)-4-fluoro-1-methylpyrrolidine-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (compound I-28 / step 3, 150 mg, 0.289 mmol) and DI EA (112 mg, 0.866 mmol) was dissolved in dichloromethane (5 mL), and (E)-4-bromo-2-enoic acid (47.6 mg, 0.289 mmol) and 2-(3H-[1,2,3]triazol[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylurammonium hexafluorophosphate (V) (110 mg, 0.289 mmol) were slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 20 °C for 1 hour, and then dimethylamine (0.866 mmol) was added to the above reaction solution and the reaction mixture was stirred for another 2 hours at room temperature. The reaction mixture was diluted with ethyl acetate and extracted. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid 2-((2S)-1-((E)-4-(dimethylamino)but-2-enyl)-4-(2'-((2S,4S)-4-fluoro-1-methylpyrrolidine-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (7.52 mg, 4.3% yield). MS: 576.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ7.33–7.02(m,4H),6.65(s,2H),5.11–4.98(m,1H),4.42–4.21(m,1H),4. 19–4.01(m,2H),3.83–3.42(m,10H),3.23–2.51(m,10H),2.45–2.13(m,10H),2.07–1.60(m,2H).

[1106] Example 36

[1107] Compound I-36: 2-((2S)-4-(2'-(((2S,4R)-4-fluoro-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile

[1108]

[1109] The synthesis of compound I-36 references compound I-14, by using (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) instead of ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro) Compound I-36 was prepared by replacing (S)-(1-methylpyrrolidone-2-yl)methanol with (2S,4R)-4-fluoro-1-methylpyrrolidone-2-yl)methanol (intermediate C3) and 2-fluoroacryloyl chloride (intermediate B1) with acryloyl chloride.

[1110] Compound I-36 (33 mg). MS: 592.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.30–7.02(m,4H),5.61–5.21(m,3H),4.75–4.50(m, 2H),4.38–3.83(m,3H),3.82–3.24(m,8H),3.24–2.59(m,9H),2.52–1.74(m,5H).

[1111] Example 37

[1112] Compound I-37

[1113] 2-((2S)-1-Acryloyl-4-(2'-(((2S,4R)-4-fluoro-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1114]

[1115] The synthesis of compound I-37 is based on compound I-14. Compound I-37 was prepared by replacing ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) with ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H-indene-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A12) and ((2S,4R)-4-fluoro-1-methylpyrrolidine-2-yl)methanol (intermediate C3) with (S)-(1-methylpyrrolidine-2-yl)methanol.

[1116] Compound I-37 (19.6 mg). MS: 574.1 (M+H) + . 1 H NMR(400MHz,Methanol-d4)δ7.17–6.98m,4H),6.81(s,1H),6.30–6.21(m,1H),5.84–5.76(m,1H) ,5.48–5.38(m,1H),4.72–3.83(m,8H),3.82–3.39(m,5H),3.24–2.59(m,9H),2.52–1.77(m,5H).

[1117] Example 38

[1118] Compound I-38: 2-((2S)-4-(2'-(((2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methoxy)-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-((E)-4-fluoro-2-enyl)piperazin-2-yl)acetonitrile

[1119]

[1120] The synthesis of compound I-38 references compound I-14, by using (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (intermediate A14) instead of ((2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-2,3-dihydro-1H) Compound I-38 was prepared by replacing (S)-(1-methylpyrrolidone-2-yl)methanol with (2S,4S)-4-fluoro-1-methylpyrrolidone-2-yl)methanol (intermediate C1) and (E)-4-fluoro-2-enoyl chloride (intermediate B5) with acryloyl chloride.

[1121] Compound I-38 (10.1 mg). MS: 606.1 (M+H) + . 1 H NMR(400MHz,DMSO-d6)δ7.29–7.01(m,4H),6.80–6.66(m,1H),5.45–5.35(m,1H),5.26–4 .90(m,1H),4.46–4.21(m,2H),3.98–3.44(m,8H),3.30–2.61(m,13H),2.31–1.58(m,6H).

[1122] Example 39

[1123] Compound II-1: 2-((2S)-1-acryloyl-4-(2'-(3-(dimethylamino)azelanoic-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1124]

[1125]

[1126] Step 1: Preparation of (2S)-2-(cyanomethyl)-4-(2-(3-(dimethylamino)azacyclobutane-1-yl)-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester

[1127]

[1128] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 600 mg, 1.026 mmol) and DIEA (1.4 mL, 8.2 mmol) were dissolved in anhydrous dichloromethane (10 mL). N,N-dimethylazacyclobutylamine hydrochloride (355 mg, 2.05 mmol) was slowly added with stirring at 25 °C. After the addition was complete, the reaction mixture was slowly raised to 50 °C and stirred for 11 hours. The reaction was then quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(2-(3-(dimethylamino)azacyclobutane-1-yl)-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (600 mg, 90.2% yield). MS: 649.1 (M+H) + .

[1129] Step 2: Preparation of (2S)-2-(cyanomethyl)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[1130]

[1131] (2S)-2-(cyanomethyl)-4-(2-(3-(dimethylamino)azacyclobutane-1-yl)-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (600 mg, 0.925 mmol) and zinc powder (181 mg, 2.77 mmol) were added to a mixed solution of acetic acid (10 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain a yellow oily substance (2S)-2-(cyanomethyl)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (480 mg, 88% yield). MS: 587.1 (M+H) + .

[1132] Step 3: Preparation of 2-((2S)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1133]

[1134] (2S)-2-(cyanomethyl)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (300 mg, 0.511 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated sodium carbonate aqueous solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 2-((2S)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (250 mg, 100% yield). MS: 487.1 (M+H) + .

[1135] Step 4: Preparation of 2-((2S)-1-acryloyl-4-(2'-(3-(dimethylamino)azelanoic-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1136]

[1137] 2-((2S)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (100 mg, 0.205 mmol) and TEA (20.79 mg, 0.205 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (18.60 mg, 0.205 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid 2-((2S)-1-acryloyl-4-(2'-(3-(dimethylamino)azelanoic-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (26 mg, 24.8% yield). MS: 541.1 (M+H) + . 1 H NMR (400MHz, Methanol-d4) δ7.21–7.03(m,4H),6.79(s,1H),6.27(d,J=16.6Hz,1H),5.81(d,J=10.6Hz,1H),4.19–3.96(m,3H), 3.97–3.68(m,4H),3.35–3.29(m,2H),3.24–2.92(m,7H),2.82–2.65(m,2H),2.21(s,6H),2.12–1.98(m,1H),1.87–1.71(m,3H).

[1138] Compounds II-1a and II-1b: 2-((S)-1-acryloyl-4-((R)-2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-acryloyl-4-((S)-2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1139]

[1140] 2-((2S)-1-acryloyl-4-(2'-(3-(dimethylamino)azelanoic-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (20 mg) was chirally resolved by SFC (column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm column, mobile phase: 40% methanol in carbon dioxide) to give a first component peak of compound II-1a (retention time 1.42 min, 5 mg) and a second component peak of compound II-1b (retention time 2.71 min, 7 mg), both white solids. MS: 541.1 (M+H) + .

[1141] Compound II-1a (retention time 1.42 min, mg). 1 H NMR (400MHz, Methanol-d4) δ7.21–7.03(m,4H),6.79(s,1H),6.27(d,J=16.6Hz,1H),5.81(d,J=10.6Hz,1H),4.19–3.96(m,3H), 3.97–3.68(m,4H),3.35–3.29(m,2H),3.24–2.92(m,7H),2.82–2.65(m,2H),2.21(s,6H),2.12–1.98(m,1H),1.87–1.65(m,3H).

[1142] Compound II-1b (retention time 2.71 minutes, 7 mg). 1 H NMR (400MHz, Methanol-d4) δ7.21–7.03(m,4H),6.79(s,1H),6.27(d,J=16.6Hz,1H),5.81(d,J=10.6Hz,1H),4.19–3.96(m,3H), 3.97–3.68(m,4H),3.35–3.29(m,2H),3.24–2.92(m,7H),2.82–2.65(m,2H),2.21(s,6H),2.12–1.98(m,1H),1.87–1.65(m,3H).

[1143] Example 40

[1144] Compound II-2: 2-((2S)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-methacryloylpiperazin-2-yl)acetonitrile

[1145]

[1146] The synthesis of compound II-2 is based on compound II-1, and compound II-2 is prepared by using 2-fluoroacryloyl chloride (intermediate B1) instead of acryloyl chloride.

[1147] Compound II-2 (34.3 mg). MS: 559.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.12–6.91(m,4H),5.29–5.19(m,2H),4.23–3.96(m,3H),3. 96–3.65(m,3H),3.42–2.91(m,11H),2.81(s,2H),2.20–2.15(m,6H),2.10–1.78(m,5H).

[1148] Compounds II-2a and II-2b: 2-((S)-4-((R)-2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile and 2-((S)-4-((S)-2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile

[1149]

[1150] 2-((2S)-4-(2'-(3-(dimethylamino)azacyclobutane-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spiro[naphthyl-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-1-methacryloylpiperazin-2-yl)acetonitrile (30 mg) was chirally resolved by SFC (column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm column, mobile phase: 40% methanol in carbon dioxide) to give a first component peak of compound II-2a (retention time 1.11 min, 10 mg) and a second component peak of compound II-2b (retention time 2.21 min, 9 mg), both white solids. MS: 559.1 (M+H) + .

[1151] Compound II-2a (retention time 1.11 min, 10 mg). 1 H NMR(400MHz, Methanol-d4)δ7.12–6.90(m,4H),5.29–4.99(m,2H),4.23–3.96(m,3H),3. 96–3.65(m,3H),3.42–2.91(m,11H),2.81(s,2H),2.20–2.12(m,6H),2.10–1.78(m,5H).

[1152] Compound II-2b (retention time 2.21 minutes, 9 mg). 1 H NMR(400MHz, Methanol-d4)δ7.12–6.90(m,4H),5.29–4.99(m,2H),4.23–3.96(m,3H),3. 96–3.65(m,3H),3.42–2.91(m,11H),2.81(s,2H),2.20–2.12(m,6H),2.10–1.78(m,5H).

[1153] Example 41

[1154] Compound II-3: 2-((2S)-1-acryloyl-4-(2'-chloro-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1155]

[1156] Step 1: Preparation of (2S)-4-(2'-chloro-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[1157]

[1158] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 250 mg, 0.427 mmol) and zinc powder (84 mg, 1.28 mmol) were added to a mixed solution of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellow solid (2S)-4-(2'-chloro-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (130 mg, 58.2% yield). MS: 524.1 (M+H) + .

[1159] Step 2: Preparation of 2-((2S)-4-(2'-chloro-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1160]

[1161] (2S)-4-(2'-chloro-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (130 mg, 0.248 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and the reaction mixture was stirred at room temperature for 11 hours. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 2-((2S)-4-(2'-chloro-6'-oxo-3,4,5',8'-tetrahydro-2H,6'-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (106 mg, 100% yield). MS: 424.1 (M+H) + .

[1162] Step 3: Preparation of 2-((2S)-1-acryloyl-4-(2'-chloro-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1163]

[1164] 2-((2S)-4-(2'-chloro-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (106 mg, 0.205 mmol) and TEA (20.79 mg, 0.205 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (18.60 mg, 0.205 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain a crude product. Pre-HPLC was performed on the crude product to prepare a white solid 2-((2S)-1-acryloyl-4-(2'-chloro-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (29.6 mg, 24.8% yield). MS: 477.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.17–6.95(m,4H),6.80(s,1H),6.28–6.12(m,1H),5.83–5.69(m,1H),5.09(s,1H),4.81–4.44 (m,1H),4.21–3.72(m,3H),3.55–3.49(m,1H),3.42–3.22(m,4H),3.18–2.96(m,1H),2.82–2.67(m,3H),2.13–1.70(m,3H).

[1165] Example 42

[1166] Compound II-4: 2-((2S)-1-acryloyl-4-(2'-hydroxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1167]

[1168] Step 1: Preparation of (2S)-2-(cyanomethyl)-4-(2'-hydroxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[1169]

[1170] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 250 mg, 0.427 mmol) and zinc powder (84 mg, 1.28 mmol) were added to a mixed solution of acetic acid (5 mL) and water (1 mL), and the reaction mixture was stirred at 75°C for 3 hours. After the reaction was complete, the reaction solution was filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a yellow solid (2S)-2-(cyanomethyl)-4-(2'-hydroxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (60 mg, 27.8% yield). MS: 505.1 (M+H) + .

[1171] Step 2: Preparation of 2-((2S)-4-(2'-hydroxy-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1172]

[1173] (2S)-2-(cyanomethyl)-4-(2'-hydroxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (60 mg, 0.119 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 2-((2S)-4-(2'-hydroxy-6'-oxo-3,4,5',8'-tetrahydro-2H,6'-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (50 mg, 100% yield). MS: 405.1 (M+H) + .

[1174] Step 3: Preparation of 2-((2S)-1-acryloyl-4-(2'-hydroxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1175]

[1176] 2-((2S)-4-(2'-hydroxy-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (48 mg, 0.119 mmol) and DIEA (33.5 mg, 0.259 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (10.74 mg, 0.119 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrously and then dried over sodium sulfate. The mixture was then filtered and concentrated to obtain a crude product. Pre-HPLC analysis of the crude product yielded a white solid, 2-((2S)-1-acryloyl-4-(2'-hydroxy-6'-oxy-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (13.6 mg, 25.1% yield). MS: 459.1 (M+H) + . 1 H NMR(400MHz, Methanol-d4)δ7.15–6.92(m,4H),6.80(s,1H),6.27–6.04(m,1H),5.82–5.65(m, 1H),4.13–3.99(m,2H),3.42–3.20(m,4H),3.16–2.69(m,5H),2.17–1.75(m,5H),1.28(s,1H).

[1177] Example 43

[1178] Compounds II-5a and II-5b: 2-((S)-1-acryloyl-4-((S)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-acryloyl-4-((R)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1179]

[1180] Step 1: Preparation of (2S)-4-(5-amino-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphth-1-yl)methyl)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester

[1181]

[1182] (2S)-4-(2-chloro-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)-5-nitropyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (intermediate A3, 180 mg, 0.308 mmol) was dissolved in methanol (5 mL). Palladium on carbon (150 mg) was added to the reaction mixture under stirring. The reaction mixture was stirred at room temperature under hydrogen for 5 hours, and then the mixture was filtered. The filtrate was concentrated under vacuum to give a colorless oil (2S)-4-(5-amino-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (130 mg, 48.7% yield). MS: 522.1 (M+H) + .

[1183] Step 2: Preparation of (2S)-2-(cyanomethyl)-4-(6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester

[1184]

[1185] (2S)-4-(5-amino-6-((1-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthyl-1-yl)methyl)pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (113 mg, 0.065 mmol) and sodium methoxide (0.436 mmol) were dissolved in DMF (2 mL), and the reaction mixture was stirred at 50°C for 1 hour. After the reaction was complete, the reaction was quenched with ice water, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was filtered and concentrated to obtain the crude product, which was purified by silica gel column chromatography to give a yellow solid (2S)-2-(cyanomethyl)-4-(6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (100 mg, 94% yield). MS: 489.1 (M+H) + .

[1186] Step 3: Preparation of 2-((2S)-4-(6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1187]

[1188] (2S)-2-(cyanomethyl)-4-(6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazine-1-carboxylic acid tert-butyl ester (100 mg, 0.205 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and evaporated to dryness, then diluted and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium carbonate solution and saturated brine, dried anhydrous and then dried over sodium sulfate. The mixture was then filtered and concentrated to give a yellow oily substance, 2-((2S)-4-(6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (80 mg, 100% yield). MS: 389.1 (M+H) + .

[1189] Step 4: Preparation of 2-((S)-1-acryloyl-4-((S)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-4'-yl)piperazin-2-yl)acetonitrile and 2-((S)-1-acryloyl-4-((R)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidine]-4'-yl)piperazin-2-yl)acetonitrile

[1190]

[1191] 2-((2S)-4-(6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic [naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile (150 mg, 0.193 mmol) and DIEA (49.9 mg, 0.386 mmol) were dissolved in dichloromethane (5 mL), and acryloyl chloride (26.2 mg, 0.290 mmol) was slowly added with stirring at 0 °C. After the addition was complete, the reaction mixture was stirred at 0 °C for 1 hour, then the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried anhydrous and then dried over sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was pre-HPLC prepared to give compounds II-5a (retention time 3.88 min, 5 mg) and II-5b (retention time 4.42 min, 8 mg), both white solids. MS: 443.1 (M+H) + .

[1192] Compound II-5a (retention time 3.88 minutes, 5 mg). 1 H NMR(400MHz,DMSO-d6)δ8.40(s,1H),7.11–6.90(m,4H),6.86(s,1H),6.22–6.13(m,1H),5.76–5.56(m,1H),5.06(s,1H),4.80(s,1H),4.3 6(s,1H),4.03(s,1H),3.59–3.49(m,2H),3.47–3.23(m,2H),3.06–2. 99(m,2H),2.95(s,2H),2.77(s,2H),2.32(s,1H),1.77–1.56(m,2H).

[1193] Compound II-5b (retention time 4.42 min, 8 mg). 1 H NMR(400MHz,DMSO-d6)δ8.40(s,1H),7.11–6.90(m,4H),6.86(s,1H),6.22–6.13(m,1H),5.76–5.56(m,1H),5.06(s,1H),4.80(s,1H),4.3 6(s,1H),4.03(s,1H),3.59–3.49(m,2H),3.47–3.23(m,2H),3.06–2. 99(m,2H),2.95(s,2H),2.77(s,2H),2.32(s,1H),1.77–1.56(m,2H).

[1194] Example 44

[1195] Compound II-6: 2-((2S)-1-acryloyl-4-(2'-(3-(diethylamino)azelanoic-1-yl)-6'-oxo-3,4,5',8'-tetrahydro-2H,6'H-spirocyclic[naphthalene-1,7'-pyridin[3,2-d]pyrimidin]-4'-yl)piperazin-2-yl)acetonitrile

[1196]

[1197] The synthesis of compound II-6 is based on compound II-1, and compound II-6 is prepared by replacing N,N-dimethylaziridine hydrochloride with N,N-diethylaziridine hydrochloride.

[1198] Compound II-6 (27.6 mg). MS: 570.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ7.24–7.01(m,4H),6.18–5.98(m,1H),5.76–5.69(m,1H),5.06(s,1H),4.78(s,1H),4.36–4.07(m ,8H),3.69–3.45(m,5H),3.13–3.07(m,4H),2.96–2.86(m,2H),2.74–2.56(m,2H),1.74–1.56(m,3H),...

Claims

1. A compound as shown in Formula I or a pharmaceutically acceptable salt thereof: Ring A is a 6-10 membered alicyclic ring containing 2 nitrogen atoms; the alicyclic ring is a monocyclic, bridged, or spirocyclic ring. R 1 -C(=O)-C(R) a )=C(R b (R) c -C(=O)-C≡CR c -C(=O)-C(=O)-OR d or -C(=O)-R d ; R a It is hydrogen, halogen or C 1-6 alkyl; R b and R c Independently hydrogen, C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 alkyl"; Each R b-1 Independent of halogen, C 1-6 Alkoxy or -NR b-2 R b-3 ;R b-2 and R b-3 Independently hydrogen or C 1-6 alkyl; R d C 1-6 alkyl; n is 0, 1, 2, or 3; Each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 alkyl"; Each R 4-1 Independently halogen, cyano, hydroxyl or C 1-6 Alkoxy; R 2 For -OR 2-1 -SR 2-2 -C(=O)R 2-3 -S(=O)2R 2-4 -S(=O)R 2-5 、-(CR 1-1 R 1-2 ) m R 2-6 or NR 1-3 R 2-7 ; R 1-1 R 1-2 and R 1-3 Independently hydrogen or C 1-6 alkyl; When m is 0, it means R 2-6 It is directly connected to the parent body via a single bond; R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and surrounded by one or more R e -1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; R 2-6 Hydrogen, CN, halogen, hydroxyl, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; R 2-7 For hydrogen, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; Each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; R e-1 R e-2 and R e-4 Independent of halogen, C 1-6 Alkyl, C 1-6 Alkoxy or NR e-5 R e-6 ;R e-5 and R e-6 Independently hydrogen or C 1-6 alkyl; Each R e-3 Independently halogen, hydroxyl, cyano, C 1-6 Alkyl, with one or more R e-7 Replacement C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R e-8 Replacement C 1-6 Alkoxy or NR e-5 R e-6 ;R e-7 and R e-8 Independently halogen, hydroxyl, cyano, C 1-6 Alkoxy or NR e-9 R e-10 R e-9 and R e-10 Independently hydrogen or C 1-6 alkyl; R 3 It is hydrogen or C 1-6 alkyl; X is O or CR 7 R 8 Y is CR 7 R 8 Z represents a single bond, O, or CR. 7 R 8 Each R 7 and R 8 Independently H or C 1-6 alkyl; U, V, W, and Q are independently CR 5 ; Each R 5 Independently H, halogen, hydroxyl group, or C substituted with one or more halogens. 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkoxy, cyano, C 3-6 cycloalkyl, C 1-6 Alkyl or C 1-6 Alkoxy; Carbon atoms marked with an asterisk (*) are chiral carbon atoms, and are in the S configuration, R configuration, or a mixture thereof.

2. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, Ring A is a 6-10 membered alicyclic ring containing 2 nitrogen atoms; the alicyclic ring is a monocyclic, bridged, or spirocyclic ring. R 1 -C(=O)-C(R) a )=C(R b (R) c -C(=O)-C≡CR c -C(=O)-C(=O)-OR d or -C(=O)-R d ; R a It is hydrogen, halogen or C 1-6 alkyl; R b and R c Independently hydrogen, C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 alkyl"; Each R b-1 Independent of halogen, C 1-6 Alkoxy or -NR b-2 R b-3 ;R b-2 and R b-3 Independently hydrogen or C 1-6 alkyl; R d C 1-6 alkyl; n is 0, 1, 2, or 3; Each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 alkyl"; Each R 4-1 Independently halogen, cyano, hydroxyl or C 1-6 Alkoxy; R 2 For -OR 2-1 -SR 2-2 -C(=O)R 2-3 -S(=O)2R 2-4 -S(=O)R 2-5 、-(CR 1-1 R 1-2 ) m R 2-6 or NR 1-3 R 2-7 ; R 1-1 R 1-2 and R 1-3 Independently hydrogen or C 1-6 alkyl; When m is 0, it means R 2-6 It is directly connected to the parent body via a single bond; R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and surrounded by one or more R e -1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; R 2-6 Hydrogen, CN, halogen, hydroxyl, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; R 2-7 For hydrogen, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl, C 1-6 alkoxy, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and surrounded by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; Each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. 10 aryl, with one or more R e-2 Replacement C6-C 10 aryl, "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms", and surrounded by one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms" and C 3-10 cycloalkyl, or, by one or more R e-4 Replacement C 3-10 cycloalkyl; R e-1 R e-2 and R e-4 Independent of halogen, C 1-6 Alkyl, C 1-6 Alkoxy or NR e-5 R e-6 ;R e-5 and R e-6 Independently hydrogen or C 1-6 alkyl; Each R e-3 Independently halogen, hydroxyl, cyano, C 1-6 Alkyl, with one or more R e-7 Replacement C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R e-8 Replacement C 1-6 Alkoxy or NR e-5 R e-6 ;R e-7 and R e-8 Independently halogen, hydroxyl, cyano, C 1-6 Alkoxy or NR e-9 R e-10 R e-9 and R e-10 Independently hydrogen or C 1-6 alkyl; R 3 It is hydrogen or C 1-6 alkyl; X is O or CH2, Y is CH2; Z is a single bond, O or CH2; U, V, W, and Q are independently CR 5 ; Each R 5 Independently H, halogen, hydroxyl group, or C substituted with one or more halogens. 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkoxy, cyano, C 3-6 cycloalkyl, C 1-6 Alkyl or C 1-6 Alkoxy; Carbon atoms marked with an asterisk (*) are chiral carbon atoms, and are in the S configuration, R configuration, or a mixture thereof.

3. The compound of formula I as described in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, Ring A is for R 2-1 C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl group, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", or a group containing one or more R groups. e-1 The substituted heteroatom is selected from one or more of N, O, and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. R 2-2 C 1-6 Alkyl or "with one or more R e Replacement C 1-6 alkyl"; R 2-5 For one or more R e Replacement C 1-6 alkyl; R 2-6 The heteroatom is hydrogen, hydroxyl, halogen, "a 5-12 membered heteroaryl group selected from one or more of N, O and S, with 1-4 heteroatoms", and is surrounded by one or more R e-1 The substituted "5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms", "4-10-membered heterocycloalkyl group selected from one or more of O and N, with 1-3 heteroatoms", or is replaced by one or more R e-3 The substituted heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms. Each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted "5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms", "4-10-membered heterocycloalkyl group selected from one or more of O and N, with 1-3 heteroatoms", or is replaced by one or more R e-3 The substituted heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms. R e-1 Independently for C 1-6 alkyl; Each R e-3 Independently halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy or NR e-5 R e-6 ; R e-5 and R e-6 Independently hydrogen or C 1-6 alkyl; R 3 It is hydrogen or C 1-6 alkyl; X is O or CR 7 R 8 Y is CR 7 R 8 Z represents a single bond, O, or CR. 7 R 8 ;R 7 and R 8 For H.

4. The compound of formula I as described in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, When R a When the halogen is halogen, the halogen is fluorine, chlorine, bromine or iodine; And / or, when R a C 1-6 When alkyl is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when R b and R c Independently for C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when each R b-1 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine or iodine; And / or, when each R b-1 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group; And / or, when R b-2 and R b-3 Independently for C 1-6 When alkyl is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when R d C 1-6 When alkyl is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when each R 4-1 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine or iodine; And / or, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 Alkyl or "with one or more R e Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group; And / or, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a group consisting of one or more R..." e-1 When the substitute is "a 5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms", then "a 5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms" is... And / or, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently defined as "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms" or "a group consisting of one or more R..." e-3 When the substituted "heteroatom selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms" is defined as follows: And / or, when R 2-6 When the halogen is halogen, the halogen is fluorine, chlorine, bromine or iodine; And / or, when R 2-6 C 1-6 Alkyl or "with one or more R e Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when R 2-6 C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group; And / or, when R 2-6 Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a group consisting of one or more R..." e-1 When the substitute is "a 5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms", then "a 5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms" is... And / or, when R 2-6 "A 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms" or "containing one or more R..." e-3 When the substituted "heteroatom selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms" is defined as follows: And / or, when each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a group consisting of one or more R..." e-1 When the substitute is "a 5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms", then "a 5-12-membered heteroaryl group selected from one or more of N, O, and S, with 1-4 heteroatoms" is... And / or, when each R e Independently defined as "a 4-10 membered heterocyclic alkyl group selected from one or more of O and N, with 1-3 heteroatoms" or "a group consisting of one or more R..." e-3 The substituted "heteroatom selected from one or more of O and N, and having 1-3 heteroatoms in a 4-10 membered heterocyclic alkyl group" refers to... And / or, when R e-1 R e-2 and R e-4 When independently a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, R e-3 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine or iodine; And / or, R e-3 Independently C1-C6 alkyl or "containing one or more R e-7 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, R e-3 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a C1-C3 alkoxy group; And / or, when R e-5 and R e-6 When independently a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when R 3 When the alkyl group is C1-C6, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when each R 5 When it is a halogen on its own, the halogen is fluorine, chlorine, bromine or iodine; And / or, when each R 5 C that is independently replaced by one or more halogens 1-6 Alkyl, the C substituted with one or more halogens 1-6 Alkyl groups are C atoms substituted with one or more halogens. 1-3 alkyl; And / or, when each R 5 Independently for C 1-6 When alkyl is used, the C1-C6 alkyl group is a C1-C3 alkyl group; And / or, when each R 5 C that is independently replaced by one or more halogens 1-6 Alkoxy, the C substituted with one or more halogens 1-6 The alkoxy group is a C-aryl group substituted with one or more halogens. 1-3 Alkoxy; And / or, when each R 7 and R 8 Independently for C 1-6 When alkyl is used, the C1-C6 alkyl group is a C1-C3 alkyl group.

5. The compound of formula I as claimed in claim 4, or a pharmaceutically acceptable salt thereof, characterized in that, for And / or, when R a When the halogen is halogen, the halogen is fluorine; And / or, when R a C 1-6 When alkyl is used, the C1-C6 alkyl group is methyl; And / or, when R b and R c Independently for C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is methyl; And / or, when each R b-1 When it is a halogen on its own, the halogen is fluorine; And / or, when each R b-1 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a methoxy group; And / or, when R b-2 and R b-3 Independently for C 1-6 When alkyl is used, the C1-C6 alkyl group is methyl; And / or, when R d C 1-6 When alkyl is used, the C1-C6 alkyl group is methyl; And / or, when each R 4 Independently for C 1-6 Alkyl or "with one or more R 4-1 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is methyl; And / or, when each R 4-1 When it is a halogen on its own, the halogen is fluorine; And / or, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 Alkyl or "with one or more R e Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is methyl; And / or, when R 2-1 R 2-2 R 2-3 R 2-4 and R 2-5 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a methoxy group; And / or, when R 2-6 When the halogen is halogen, the halogen is chlorine; And / or, when R 2-6 C 1-6 Alkyl or "with one or more R e Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is methyl; And / or, when R 2-6 C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a methoxy group; And / or, when R e-1 R e-2 and R e-4 When independently a C1-C6 alkyl group, the C1-C6 alkyl group is methyl; And / or, R e-3 When it is a halogen on its own, the halogen is fluorine; And / or, R e-3 Independently C1-C6 alkyl or "containing one or more R e-7 Replacement C 1-6 When "alkyl" is used, the C1-C6 alkyl group is methyl; And / or, R e-3 Independently for C 1-6 When alkoxy is used, the C1-C6 alkoxy group is a methoxy group; And / or, when R e-5 and R e-6 When independently a C1-C6 alkyl group, the C1-C6 alkyl group is methyl or ethyl; And / or, when R 3 When the alkyl group is C1-C6, the C1-C6 alkyl group is methyl; And / or, when each R 5 When it is a halogen on its own, the halogen is fluorine, chlorine, or bromine; And / or, when each R 5 C that is independently replaced by one or more halogens 1-6 Alkyl, the C substituted with one or more halogens 1-6 The alkyl group is -CF3; And / or, when each R 5 Independently for C 1-6 When alkyl is used, the C1-C6 alkyl group is methyl; And / or, when each R 5 C that is independently replaced by one or more halogens 1-6 Alkoxy, the C substituted with one or more halogens 1-6 The alkoxy group is -OCF3; And / or, when each R 7 and R 8 Independently for C 1-6 When alkyl is used, the C1-C6 alkyl group is methyl.

6. The compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, Ring A is a 6-10 membered alicyclic heterocycle containing 2 N atoms, and the alicyclic heterocycle is a monocyclic ring; And / or, R 1 -C(=O)-C(R) a )=C(R b (R) c ), -C(=O)-C≡CCH3, -C(=O)-C(=O)-OCH3 or -C(=O)-CH3; when R a Halogen or C 1-6 When alkyl, R b and R c It is hydrogen; when R b or R c C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 When "alkyl", R a It is hydrogen; And / or, R a It is hydrogen or halogen; And / or, R b and R c Independently hydrogen, C 1-6 Alkyl or "with one or more R b-1 Replacement C 1-6 "alkyl", and R b and R c At least one of them is hydrogen; And / or, each R b-1 Halogens are independent of each other; And / or, n is 0, 1 or 2; And / or, each R 4 Methyl group that is independently substituted with methyl, cyano, F, or hydroxy; And / or, R 2-1 For hydrogen, C 1-6 Alkyl, with one or more R e Replacement C 1-6 Alkyl or "with one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms. And / or, R 2-2 C 1-6 Alkyl or "with one or more R e Replacement C 1-6 alkyl"; And / or, R 2-5 For one or more R e Replacement C 1-6 alkyl; And / or, R 2-6 Hydrogen, halogen, or one or more R e-3 The substituted "a heteroatom selected from one or more of O and N, and a 4-10 membered heterocyclic alkyl group having 1-3 heteroatoms", "a heteroatom selected from one or more of N, O and S, and a 5-12 membered heteroaryl group having 1-4 heteroatoms", or substituted by one or more R e-1 The substituted heteroatom is selected from one or more of N, O and S, and is a 5-12 membered heteroaryl group with 1-4 heteroatoms or a hydroxyl group; And / or, each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", and constituting one or more R e-1 The substituted "5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms", "4-10-membered heterocycloalkyl group with one or more heteroatoms selected from O and N, and having 1-3 heteroatoms", or "substituted by one or more R..." e-3 The substituted heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms. And / or, R e-1 Independently for C 1-6 alkyl; And / or, each R e-3 Independently halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy or NR e-5 R e-6 ; And / or, R 2 For -OR 2-1 -SR 2-2 -S(=O)R 2-5 or -(CR) 1-1 R 1-2 ) m R 2-6 ; And / or, X is O or CR 7 R 8 Y is CR 7 R 8 Z represents a single bond, O, or CR. 7 R 8 ; And / or, each R 5 Independently, H, halogen, or C substituted with one or more halogens. 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkoxy, cyano, C 3-6 cycloalkyl, C 1-6 Alkyl or C 1-6 Alkyl group.

7. The compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, R b and R c Independently hydrogen or "C substituted with one or more halogens" 1-6 "alkyl", and R b and R c At least one of them is hydrogen; And / or, n is 1 or 2; And / or, each R 4 A methyl group that is independently substituted with a cyano group; And / or, each R e-3 Independently halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy or NR e-5 R e-6 ; And / or, each R e Independently defined as "a 5-12-membered heteroaryl group with one or more heteroatoms selected from N, O, and S, and having 1-4 heteroatoms" or "a group consisting of one or more R..." e-3 The substituted heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms. And / or, X and Y are CH2, and Z is a single bond, O, or CH2; And / or, each R 5 Independent of H, halogen or C 1-6 alkyl.

8. The compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, for And / or, R 1 for And / or, R 2 For -OR 2-1 or -(CR) 1-1 R 1-2 ) m R 2-6 R 2-1 For one or more R e Replacement C 1-6 Alkyl groups, each R e Independently for one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", R e In the middle, each R e-3 Independent of halogen, C 1-6 Alkyl or C 1-6 Alkoxy group; m is 0, R 2-6 For one or more R e-3 The substituted "heteroatom is selected from one or more of O and N, and is a 4-10 membered heterocyclic alkyl group with 1-3 heteroatoms", R 2-6 In the middle, each R e-3 Independent for NR e-5 R e-6 .

9. The compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, for And / or, R 2 For hydrogen, hydroxyl, chlorine, -OCH3, -SCH3 And / or, for 10. The compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, The compound represented by Formula I is any one of the following compounds:

11. The compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that, The compound represented by Formula I is any one of the following compounds: The compound with a retention time of 1.21 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.99 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 0.93 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.43 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.24 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 2.56 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.01 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 2.17 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.74 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 5.44 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.08 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.22 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.42 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 2.71 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.11 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 2.21 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 3.88 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.42 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 1.254 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 2.267 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.473 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.688 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound that has a retention time of 7.008 min under the following conditions is: One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 7.199 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.735 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.758 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 7.198 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 7.460 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.249 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.333 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 6.961 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 7.133 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 4.903 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 5.508 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 2.174 min under the following conditions is... One of the stereoisomers: Column: CHIRALART Cellulose SB, 2cm×25cm, 5μm; Mobile phase A: MtBE, MtBE contains 0.5% by volume of 2mM NH3 in MEOH, Mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 2.950 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALART Cellulose SB, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 1.986 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALPAK IE, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 2.919 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALPAK IE, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 5.137 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 5.370 min under the following conditions is... One of the stereoisomers: Column: Dr. Maish Reprosil Chiral-JM, 250 x 25 mm, 10 μm; Column temperature: room temperature; Mobile phase: CO2 / MeOH = 60 / 40; The compound with a retention time of 0.928 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALPAK IF, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 1.411 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALPAK IF, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 1.872 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE, Mobile phase B: MeOH:DCM=1:1; The compound with a retention time of 2.467 min under the following conditions is... One of the stereoisomers: Column: CHIRALPAK IF, 2cm×25cm, 5μm; Mobile phase A: MtBE, Mobile phase B: MeOH:DCM=1:1; The compound with a retention time of 0.993 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALPAK IF, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B: MeOH:DCM = 1:1; The compound with a retention time of 2.596 min under the following conditions is... One of the stereoisomers: chromatographic column: CHIRALPAK IF, 2cm×25cm, 5μm; mobile phase A: MtBE, MtBE containing 0.5% by volume of 2mM NH3 in MEOH solution, mobile phase B:MeOH:DCM=1:

1.

12. A method for preparing a compound of formula I as described in any one of claims 1 to 11, characterized in that, It includes the following steps: The compounds shown in formula KX and K-XIII undergo substitution reactions to yield the compound shown in formula I. Among them, "*", n, ring A, R 1 R 2 R 3 R 4 The definitions of X, Y, Z, U, V, W and Q are as described in any one of claims 1 to 11, where L is a halogen or a hydroxyl group.

13. The method for preparing the compound as shown in Formula I according to claim 12, characterized in that, The method for preparing the compound as shown in Formula I further includes the following steps: The compound shown in formula K-IX is deprotected to give the compound shown in formula KX; Among them, "*", n, ring A, R 2 R 3 R 4 The definitions of X, Y, Z, U, V, W and Q are as described in any one of claims 1 to 10, where PG is an amino protecting group.

14. A compound as shown in formula KX: in, *, n, ring A, R 2 R 3 R 4 The definitions of X, Y, Z, U, V, W, and Q are as described in any one of claims 1 to 11.

15. The compound of formula KX as claimed in claim 14, characterized in that, The compound represented by formula KX is any of the following compounds:

16. A pharmaceutical composition comprising a compound of Formula I as described in any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

17. The use of a compound of Formula I as described in any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 16, in the preparation of a medicament.

18. The application as described in claim 17, characterized in that, The drug is used to treat cancer.

19. The application as described in claim 18, characterized in that, The cancers mentioned include lung cancer, colon cancer, pancreatic cancer, rectal cancer, lymphoma, esophageal cancer, ovarian cancer, glioma, cervical cancer, urothelial carcinoma, stomach cancer, endometrial cancer, liver cancer, bile duct cancer, breast cancer, leukemia, and melanoma.

20. The compound of Formula I as described in any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 16, in the preparation of KRAS G12C Applications in protein mutation inhibitors.

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