PKMYT1 inhibitor, preparation method, pharmaceutical composition and application of PKMYT1 inhibitor

Abstract

The invention belongs to the field of medicines, and discloses a PKMYT1 inhibitor, a preparation method, a pharmaceutical composition and application thereof. The invention provides a compound as shown in a general formula (I), and pharmaceutically acceptable salts or isomers thereof. The PKMYT1 inhibitor provided by the invention can be used for treating neoplastic diseases caused by CCNE1 amplification or FBXW7 inactivation mutation. # imgabs0 #

Description

PKMYT1 inhibitor, preparation method, pharmaceutical composition and use thereof

[0001] This application claims priority to Chinese Patent Application No. 2022114881146 filed on November 25, 2022, Chinese Patent Application No. 202310519621X filed on May 9, 2023, Chinese Patent Application No. 2023111342853 filed on September 4, 2023, and Chinese Patent Application No. 2023114943221 filed on November 9, 2023. The entire contents of the aforementioned Chinese patent applications are incorporated herein by reference. Technical Field

[0002] The present invention belongs to the field of medicine, and specifically relates to a PKMYT1 inhibitor compound, a pharmaceutically acceptable salt thereof, an isomer thereof, a preparation method thereof, a pharmaceutical composition thereof, and medical uses thereof. Background Art

[0003] Cyclin E is a cyclin of the cyclin-dependent kinase 2 (CDK2) family. It binds to CDK2 in the G1 phase to form an activated CDK2-Cyclin E complex, promoting the transition from the G1 phase to the S phase of the cell cycle, where DNA replication initiates. The Cyclin E1 (CCNE1) gene encodes the primary protein of Cyclin E and thus plays a crucial role in regulating the G1-S phase transition of the cell cycle. Studies have suggested that overexpression of CCNE1 protein can lead to increased expression of the encoded Cyclin E protein, thereby enhancing the activity of the CDK2-Cyclin E complex, triggering premature cell cycle transitions, increasing DNA replication stress, and genomic instability. Amplification of CCNE1 is prevalent in various tumor types, particularly in highly malignant gynecological and gastrointestinal cancers, such as ovarian cancer (HGSOC), uterine cancer, and gastroesophageal cancer, and is associated with resistance to cytotoxic and targeted therapies.

[0004] CCNE1 itself is not considered a druggable target. Current research focuses on multi-targeted CDK inhibitors that act on its downstream cyclin, CDK2. Therefore, the lack of treatment options for CCNE1-amplified tumors makes the development of new therapies for this type of tumor a critical unmet need. To identify therapeutic targets for CCNE1-amplified tumors, a collaboration between Mount Sinai Hospital, the University of Toronto, and Repare Therapeutics in the United States discovered that CCNE1 amplification and PKMYT1 inhibition constitute a synthetic lethal pair. This finding was published in Nature on April 20, 2022. They conducted a genome-scale CRISPR-Cas9-based synthetic lethality screen in a cell model of CCNE1 amplification. Their results revealed that PKMYT1 is essential in CCNE1-amplified cells but not in otherwise healthy cells with normal CCNE1 levels. Therefore, PKMYT1 is a synthetic lethal gene for CCNE1, and PKMYT1 inhibitors could be used to treat CCNE1-amplified tumors. Furthermore, the protein encoded by the FBXW7 gene is the target protein recognition component of the cullin-RING ubiquitin ligase. The FBXW7 protein targets CCNE1 through the ubiquitin-dependent protein degradation pathway. Therefore, FBXW7 inactivation mutations will lead to increased CCNE1 levels. PKMYT1 inhibitors can also be used to treat tumor diseases with FBXW7 inactivation mutations.

[0005] PKMYT1 kinase, also known as MYT1, whose full name is "Membrane-associated tyrosine and threonine-specific cdc2 inhibitory kinase", is a member of the WEE family of kinases. During cell cycle transitions, it phosphorylates the threonine 14 site of CDK1 kinase, rendering the CDK1-Cyclin B complex inactive, negatively regulating the cell cycle checkpoints from G2 to M, and has an important impact on tumor cell proliferation, migration, and xenograft tumor formation.

[0006] Based on the discovery of a synthetic lethal relationship between CCNE1 and PKMYT1, a research team at Mount Sinai Hospital in Toronto developed the selective PKMYT1 inhibitor RP-6306. When used in combination with gemcitabine in a CCNE1 amplification model, RP-6306 demonstrated single-agent activity and durable tumor regression. RP-6306 treatment resulted in selective, unplanned activation of CDK1 in CCNE1-overexpressing cells, promoting premature mitosis in cells undergoing DNA synthesis. CCNE1 overexpression disrupts CDK1 homeostasis, at least in part, through premature activation of the MMB-FOXM1 mitotic transcriptional program. They concluded that PKMYT1 inhibition is a promising strategy for treating CCNE1-amplified cancers.

[0007] Currently, RP-6306 is still in the clinical stage, and there are only a few drug options available. Therefore, it is still of great clinical significance to continue to develop selective PKMYT1 inhibitors, enrich the variety of drugs, and increase drug accessibility.

[0008] Summary of the Invention

[0009] The present disclosure aims to address one or more of the shortcomings of existing PKMYT1 inhibitors, including limited availability, low PKMYT1 enzyme inhibitory activity, low HCC1569 cell inhibitory activity, short half-life, rapid clearance rate, poor metabolic stability, poor anti-tumor activity, and poor solubility. The present invention provides a novel PKMYT1 inhibitor, which exhibits one or more advantages, including high PKMYT1 enzyme inhibitory activity, high HCC1569 cell inhibitory activity, long half-life, slow clearance rate, good metabolic stability, good anti-tumor activity, and good solubility. The inhibitor can be used to treat neoplastic diseases characterized by CCNE1 amplification or FBXW7 inactivating mutations.

[0010] The present invention mainly solves the above technical problems through the following technical solutions.

[0011] In a first aspect, the present disclosure provides a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof:

[0012] in,

[0013] X 1 Select N, or CR 5 ;

[0014] X 2 Select N, or CR 6 ;

[0015] X 3 Select N, or CR 7 ;

[0016] X 4 Select N, or CR 8 ;

[0017] X 5 Selected from N, or C;

[0018] X 6 Selected from N, or C;

[0019] X 7 Selected from N, or C;

[0020] The condition is that when X 2 Selected from CR 6 , and X 5Selected from C, and X 6 Select from N, and X 7 When selected from C, X 3 and X 4 Not at the same time CH;

[0021] R 1 and R 2 are independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy C 1-4 Alkyl or deuterated C 1-4 alkyl;

[0022] R 3 Selected from -H, -CN, -OH, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclic group, 3-6 membered heterocycloalkenyl, deuterated C 1-4 alkyl, or 5-6 membered heteroaryl; wherein the phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 3a Substituted, the R 3a Selected from halogen, C 1-4 Alkyl, or C 1-4 alkoxy;

[0023] L is selected from C 2-4 Alkynylidene, C 1-4 Alkylene, or C 2-4 alkenylene;

[0024] R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, and 4-6 membered heterocyclic groups are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH;

[0025] R a and R b are independently selected from -H, C 1-4Alkyl, phenyl, p-methoxybenzyl, C 1-4 Alkyl-C(O)-, C 3-6 Cycloalkyl, or halogenated C 1-4 Alkyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen;

[0026] R 4 Selected from -H, halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl;

[0027] R 5 、R 7 , and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl;

[0028] R 6 Selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy, -COOH, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl-S(O)2-, hydroxyl C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3- 6-cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, 5-6 membered heteroaryl, or -CD3; wherein the C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3-6 Cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 6a Substituted, the R 6aSelected from halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, or C 1-4 alkoxy;

[0029] R d and R e are independently selected from -H, C 1-4 Alkyl, -S(O)2-N(R f )(R g ), -C(O)-aryl, -C(O)-NR f -aryl, or -(C=S)-NR f -aryl;

[0030] R f and R g are independently selected from -H, or C 1-4 alkyl;

[0031] Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 cycloalkenyl;

[0032] and / or, R 3 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl;

[0033] and / or, R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl;

[0034] The phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkenyl, and C 5-6 Cycloalkenyl is optionally further substituted by one or more alkyl radicals selected from halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-6 The alkyl-C(O)- group is substituted.

[0035] In another aspect, the present disclosure provides a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0036] in,

[0037] X 1 Select N, or CR 5 ;

[0038] X2 Select N, or CR 6 ;

[0039] X 3 Select N, or CR 7 ;

[0040] X 4 Select N, or CR 8 ;

[0041] X 5 Selected from N, or C;

[0042] X 6 Selected from N, or C;

[0043] X 7 Selected from N, or C;

[0044] The condition is that when X 2 Selected from CR 6 , and X 5 Selected from C, and X 6 Select from N, and X 7 When selected from C, X 3 and X 4 Not at the same time CH;

[0045] R 1 and R 2 are independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy C 1-4 Alkyl or deuterated C 1-4 alkyl;

[0046] R 3 Selected from -H, -CN, -OH, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclic group, 3-6 membered heterocycloalkenyl, deuterated C 1-4 alkyl, or 5-6 membered heteroaryl; wherein the phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 3a Substituted, the R 3a Selected from halogen, C 1-4Alkyl, or C 1-4 alkoxy;

[0047] L is selected from C 2-4 Alkynylidene, C 1-4 Alkylene, or C 2-4 alkenylene;

[0048] R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, and 4-6 membered heterocyclic groups are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH;

[0049] R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, p-methoxybenzyl, C 1-4 Alkyl-C(O)-, C 3-6 Cycloalkyl, or halogenated C 1-4 Alkyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen;

[0050] R 4 Selected from -H, halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl;

[0051] R 5 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl;

[0052] R 6 Selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy, -COOH, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 2-4 Alkenyl, C2-4 Alkynyl, C 1-4 Alkyl-S(O)2-, hydroxyl C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3- 6-cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, 5-6 membered heteroaryl, or -CD3; wherein the C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3-6 Cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 6a Substituted, the R 6a Selected from halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl or C 1-4 alkoxy;

[0053] R d and R e are independently selected from -H, C 1-4 Alkyl, -S(O)2-N(R f )(R g ), -C(O)-aryl, -C(O)-NR f -aryl, or -(C=S)-NR f -aryl;

[0054] R f and R g are independently selected from -H, or C 1-4 alkyl;

[0055] Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 cycloalkenyl;

[0056] and / or, R 3 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl;

[0057] and / or, R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl;

[0058] The phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkenyl, and C 5-6 Cycloalkenyl is optionally further substituted by one or more alkyl radicals selected from halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl and C 1-4 substituted by an alkoxy substituent.

[0059] In some embodiments,

[0060] X 1 Select N, or CR 5 ;

[0061] X 2 Select N, or CR 6 ;

[0062] X 3 Select N, or CR 7 ;

[0063] X 4 Select N, or CR 8 ;

[0064] X 5 Selected from N, or C;

[0065] X 6 Selected from N, or C;

[0066] X 7 Selected from N, or C;

[0067] The condition is that when X 2 Selected from CR 6 , and X 5 Selected from C, and X 6 Select from N, and X 7 When selected from C, X 3 and X 4 Not at the same time CH;

[0068] R 1 and R 2 are independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy or hydroxy C 1-4 alkyl;

[0069] R 3 Selected from -H, -CN, -OH, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 cycloalkenyl, 3-6 membered heterocyclic group, or 3-6 membered heterocyclic alkenyl; wherein the phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclyl, and 3-6 membered heterocycloalkenyl are optionally substituted with one or more R 3a Substituted, the R 3a Selected from halogen, C 1-4 Alkyl or C 1-4 alkoxy;

[0070] L is selected from C 2-4 Alkynylidene, C 1-4 Alkylene, or C 2-4 alkenylene;

[0071] R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH;

[0072] R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, p-methoxybenzyl, C 1-4 Alkyl-C(O)-, C 3-6 Cycloalkyl, or halogenated C 1-4 Alkyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen;

[0073] R 4 Selected from -H, halogen, C 1-4 Alkyl, or C 3-6 Cycloalkyl;

[0074] R 5 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl;

[0075] R 6 Selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy, -COOH, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl-S(O)2-, hydroxyl C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3- 6-cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, or 5-6 membered heteroaryl; wherein the C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3-6 Cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 6a Substituted, the R 6a Selected from halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, or C 1-4 alkoxy;

[0076] R d and R e are independently selected from -H, C 1-4 Alkyl, -S(O)2-N(R f )(R g ), -C(O)-aryl, -C(O)-NR f -aryl, or -(C=S)-NR f -aryl;

[0077] R f and R g are independently selected from -H, or C 1-4 alkyl;

[0078] Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 cycloalkenyl;

[0079] and / or, R 3 、R6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, or a C 5-6 cycloalkenyl;

[0080] and / or, R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, or a C 5-6 Cycloalkenyl.

[0081] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer,

[0082] in,

[0083] X 1 Select N, or CR 5 ;

[0084] X 2 Select N, or CR 6 ;

[0085] X 3 Select N, or CR 7 ;

[0086] X 4 Select N, or CR 8 ;

[0087] X 5 Selected from N, or C;

[0088] X 6 Selected from N, or C;

[0089] X 7 Selected from N, or C;

[0090] The condition is that when X 2 Selected from CR 6 , and X 5 Selected from C, and X 6 Select from N, and X 7 When selected from C, X 3 and X 4 Not at the same time CH;

[0091] R 1 and R 2 are independently selected from C 1-4 alkyl;

[0092] R 3 Selected from -H, -CN, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 3-6 membered cycloalkenyl, 3-6 membered heterocyclyl, or 3-6 membered heterocycloalkenyl;

[0093] L is selected from C 2-4 Alkynylidene;

[0094] R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, and 4-6 membered heterocyclic groups are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH;

[0095] R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, or p-methoxybenzyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen;

[0096] R 4 Selected from -H, halogen, or C 1-4 alkyl;

[0097] R 5 、R 6 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, or C 1-4 alkoxy;

[0098] R d and R e are independently selected from -H, or C 1-4 alkyl;

[0099] or R 2 、R 8 Together with the atoms it is connected to form C 5-6 Cycloalkenyl.

[0100] In another aspect, the present invention provides a compound represented by the following general formula (I):

[0101] in,

[0102] X 1 Select N, or CR 5 ;

[0103] X 2 Select N, or CR 6 ;

[0104] X 3 Select N, or CR 7 ;

[0105] X 4 Select N, or CR 8 ;

[0106] X 5 Selected from N, or C;

[0107] X 6 Selected from N, or C;

[0108] X 7 Selected from N, or C;

[0109] The condition is that when X 2 Selected from CR 6 , and X 5 Selected from C, and X 6 Select from N, and X 7 When selected from C, X 3 and X 4 Not at the same time CH;

[0110] R 1 and R 2 are independently selected from C 1-4 alkyl;

[0111] R 3 Selected from -H, -CN, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkenyl, C 1-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 3-6 membered cycloalkenyl, 3-6 membered heterocyclyl, or 3-6 membered heterocycloalkenyl;

[0112] L is selected from C 1-4 Alkynylidene;

[0113] R c Selected from C 1-4Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclyl are optionally substituted by halogen, or -OH;

[0114] R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, or p-methoxybenzyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen;

[0115] R 4 Selected from -H, halogen, or C 1-4 alkyl;

[0116] R 5 、R 6 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, or C 1-4 alkoxy;

[0117] R d and R e are independently selected from -H, or C 1-4 alkyl;

[0118] or R 2 、R 8 Together with the atoms to which it is attached, it forms a 5-6 membered cycloalkenyl.

[0119] In a preferred embodiment of formula (I), X 1 Selected from CR 5 , X 2 Selected from N, X 5 Selected from C, X 6 Selected from N, X 7 Selected from C.

[0120] In a preferred embodiment of formula (I), X 1 and X 2 All selected from N, X 5 Selected from C, X 6 Selected from N, X 7 Selected from C.

[0121] In a preferred embodiment of formula (I), R 1 and R 2Each is independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH or -CD3.

[0122] In a preferred embodiment of formula (I), R 1 and R 2 Each is independently selected from -CH3, -CH2CH3, or -CH(CH3)2.

[0123] In a preferred embodiment of formula (I), R 1 and R 2 Each is independently selected from -Cl, -F, -Br, -OCH3, or -CH2OH.

[0124] In a preferred embodiment of formula (I), R 1 and R 2 are all selected from -CH3.

[0125] In a preferred embodiment of formula (I), R 1 and R 2 Each is independently selected from -CD3.

[0126] In a preferred embodiment of formula (I), R 1 and R 2 are selected from -CH3 or -CD3.

[0127] In a preferred embodiment of formula (I), R 3 Selected from -H, -CN, -NH2, -F, -Br, -Cl, -CH3, -CH2CH3, -CH2CH(CH3)2, -CF3, -OCH3, -OCH2CH3, -CH=CH2, -N(CH3)2, -N(CH2CH3)2, -NHPMB, phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Ethylene, -C=CR c , -OH, -CH(CH3)2, -CH2CH2CH3, -CH2CH(CH3)2, -OCH3, -CH2F, -CHF2, -CH=CH2, -CH=C(CH3)2, -CH=CHCH3, -C(CH3)=CH2, -NHCH3, -NHCH2CH3, -NHC(O)CH3, or-CD3,R c Selected from optionally one or more R ca The following groups are substituted: methyl, ethyl, cyclopropyl, cyclobutyl, isopropyl, oxetanyl, or azetidine, wherein R ca Selected from -F, -Cl, or -OH.

[0128] In a preferred embodiment of formula (I), R 3 Selected from -H, -CN, -NH2, -F, -Br, -Cl, -CH3, -CH2CH3, -CH2CH(CH3)2, -CF3, -OCH3, -OCH2CH3, -CH=CH2, -N(CH3)2, -N(CH2CH3)2, -NHPMB, phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Ethylene, or -C=CR c , R c Selected from optionally one or more R ca The following groups are substituted: methyl, ethyl, cyclopropyl, cyclobutyl, isopropyl, oxetanyl, or azetidine, wherein R ca Selected from -F, -Cl, or -OH.

[0129] In a preferred embodiment of formula (I), R 3 Selected from -OH, -CH(CH3)2, -CH2CH2CH3, -CH2CH(CH3)2, -OCH3, -CH2F, -CHF2, -CH=CH2, -CH=C(CH3)2, -CH=CHCH3, -C(CH3)=CH2, -NHCH3, -NHCH2CH3, -NHC(O)CH3,

[0130] In a preferred embodiment of formula (I), R 3 Selected from or -CD3.

[0131] In a preferred embodiment of formula (I), -C≡CR c Selected from -C=C-CH3,

[0132] In a preferred embodiment of formula (I), R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, cyclopropyl, methoxy, or -CH2CF3.

[0133] In a preferred embodiment of formula (I), R 4 Selected from -H, -Cl, -F, -Br, -CH3, or -CH2CH3.

[0134] In a preferred embodiment of formula (I), R 4 Selected from cyclopropyl.

[0135] In a preferred embodiment of formula (I), R 4 Selected from -H.

[0136] In a preferred embodiment of formula (I), R 4 Selected from methoxy, or -CH2CF3.

[0137] In a preferred embodiment of formula (I), wherein R 5 Selected from -H, -Cl, -F, -CH3, -CN, -CF3, or -OCH3.

[0138] In a preferred embodiment of formula (I), wherein R 5 Selected from -H, -Cl, -F, -CH3, or -OCH3.

[0139] In a preferred embodiment of formula (I), wherein R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -F, -Br, -Cl, -CH3, -CH2CH3, -OCH3, or -OCH2CH3.

[0140] In a preferred embodiment of formula (I), R 7 and R 8 Both are H.

[0141] In a preferred embodiment of formula (I), R 7 and R 8 All are F.

[0142] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has any of the following structures:

[0143] Among them, X 1 、X 2 、R 1 、R 2 、R 3 、R 4 、R 5 、R 7 and R 8 As defined in any of the above schemes.

[0144] In another preferred embodiment of formula (I), X 1 Select from N;X 2 Selected from CR 6 ;X 3 Select N, or CR 7 ;X 5 Selected from C;X 6 Select from N;X 7 Selected from C, and X3 and X 4 Not CH at the same time.

[0145] In another preferred embodiment of formula (I), X 1 Select from N;X 2 Selected from CR 6 ;X 5 Selected from C;X 6 Select from N;X 7 Selected from C;X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 , and X 3 and X 4 Not CH at the same time.

[0146] In a preferred embodiment of formula (I), R 1 and R 2 Each is independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH or -CD3; preferably, R 1 and R 2 are all selected from -CH3 or -CD3;

[0147] R 3 Selected from -CH3, -NH2, -OH, -F, -Cl, -Br, -CH(CH3)2, -CH2CH3, -CH2CH2CH3, -CH2CH(CH3)2, -OCH3, -CH2F, -CHF2, -CF3, -CH=CH2, -CH=C(CH3)2, -CH=CHCH3, -C(CH3)=CH2, -NHCH3, -NHCH2CH3, -NHC(O)CH3, or -CD3;

[0148] R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, cyclopropyl, or -CH2CF3;

[0149] R 6Selected from -CH3, -H, -OH, -Cl, -F, -Br, -COOH, -CN, -CH2CH3, -CH(CH3)2, -CH2CH(CH3)2, -CF3, -CHF2, -CH2F, -CF2C H3, -CF(CH3)2, -OCH3, -OCH(CH3), -OCF3, -OCH2CF3, -N(CH3)2, -CH=CH2, -C(CH3)=CH2, -C≡CH, -S(O)2CH3, -CH2CH2CH=CH2, -OCH(CH3)2, -CH2OH, -CH2CF3, or -CD3;

[0150] X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ;

[0151] R 7 and R 8 are independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3; and R 7 and R 8 Not at the same time H;

[0152] Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 Cycloalkenyl.

[0153] In a preferred embodiment of formula (I), R 1 and R 2 are independently selected from -CH3, -CH2CH3, or -CH(CH3)2; preferably, R 1 and R 2 are all selected from -CH3;

[0154] R 3 Selected from -CH3;

[0155] R 4 Selected from -H;

[0156] R 6 Selected from -CH3;

[0157] X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 , and X 3 and X4 Not at the same time CH;

[0158] R 7 and R 8 Each independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -OCH3, or -OCH2CH3;

[0159] R 7 and R 8 Not at the same time H;

[0160] or R 2 、R 8 Together with the atoms it is connected to form C 5-6 Cycloalkenyl.

[0161] In a preferred embodiment of formula (I), X 3 CR 7 , X 4 CR 8 ; R 7 and R 8 R is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -OCH3, or -OCH2CH3; 7 and R 8 Not H at the same time.

[0162] In a preferred embodiment of formula (I), R 1 and R 2 Each is independently selected from -Cl, -F, -Br, -OCH3, or -CH2OH.

[0163] In a preferred embodiment of formula (I), R 1 and R 2 Each is independently selected from -CD3.

[0164] In a preferred embodiment of formula (I), R 3 Selected from -NH2, -OH, -F, -Cl, -Br, -CH(CH3)2, -CH2CH3, -CH2CH2CH3, -CH2CH(CH3)2, -OCH3, -CH2F, -C HF2, -CF3, -CH=CH2, -CH=C(CH3)2, -CH=CHCH3, -C(CH3)=CH2, -NHCH3, -NHCH2CH3, -NHC(O)CH3,

[0165] In a preferred embodiment of formula (I), R 3 Selected from or -CD3.

[0166] In a preferred embodiment of formula (I), R 4 Selected from -Cl, -F, -Br, -CH3, -CH2CH3, or cyclopropyl.

[0167] In a preferred embodiment of formula (I), R 4 Selected from -CH2CF3.

[0168] In a preferred embodiment of formula (I), R 6 Selected from -H, -OH, -Cl, -F, -Br, -COOH, -CN, -CH2CH3, -CH(CH3)2, -CH2CH(CH3)2, -CF3, -CHF2, -CH2F, -CF2CH3, -CF(CH3)2, -OCH3, -OCH(CH3), -OCF3, -OCH2CF3, -N(CH3)2, -CH=CH2, -C(CH3)=CH2, -C≡CH, -S(O)2CH3,

[0169] In a preferred embodiment of formula (I), R 6 Selected from -CH2CH2CH=CH2, -OCH(CH3)2, -CH2OH, -CH2CF3, or -CD3.

[0170] In a preferred embodiment of formula (I), R 7 and R 8 are each independently selected from -Br.

[0171] In a preferred embodiment of formula (I), R 7 and R 8 are independently selected from -H, or -F, and R 7 and R 8 At least one is -F.

[0172] In a preferred embodiment of formula (I), X 3 Selected from CR 7 , X 4 Selected from CR 8 ;

[0173] R 1 and R 2 Selected from C 1-4 Alkyl, or deuterated C 1-4 Alkyl; preferably, R 1 , and R 2 Selected from -CH3 or -CD3;

[0174] R 3 Selected from C 1-4 Alkyl; preferably, R 3 Selected from -CH3 or -CH2CH3;

[0175] R 4 Selected from -H;

[0176] R 6 Selected from halogen, or halogenated C 1-4 Alkyl; preferably, R 6 Selected from -Cl, -Br, or -CF3;

[0177] R 7 and R 8 are independently selected from -H or halogen, and R 7 and R 8 are not hydrogen at the same time; preferably, R 7 and R 8 are independently selected from -H or F, and R 7 and R 8 Not hydrogen at the same time.

[0178] In a preferred embodiment of formula (I), X 1 Select from N;X 2 Selected from CR 6 ;X 3 Selected from CR 7 ;X 4 Selected from CR 8 ;X 5 Selected from C;X 6 Select from N;X 7 Selected from C;

[0179] R 1 and R 2 are independently selected from C 1-4 Alkyl, preferably -CH3;

[0180] R 3 Selected from C 1-4 Alkyl, preferably -CH3;

[0181] R 4 Selected from -H;

[0182] R 7 is selected from halogen, preferably F; R 8 Selected from -H;

[0183] R 6 Selected from halogen or halogenated C 1-4 The alkyl group is preferably Br or -CF3, more preferably -CF3.

[0184] In a preferred embodiment of formula (I), R1 and R 2 are independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH, or -CD3; preferably, R 1 and R 2 are all selected from -CH3, or -CD3;

[0185] R 3 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0186] R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, or cyclopropyl;

[0187] X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ;

[0188] R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

[0189] In a preferred embodiment of formula (I), wherein R 1 and R 2 are independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3 or -CH2OH; preferably, R 1 and R 2 are all selected from -CH3;

[0190] R 3 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0191] R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, or cyclopropyl;

[0192] X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ;

[0193] R 7 and R8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

[0194] In a preferred embodiment of formula (I), R 1 and R 2 are independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH, or -CD3; preferably, R 1 and R 2 are all selected from -CH3, or -CD3;

[0195] R 3 is selected from -H, -CH3, or -CH2CH3;

[0196] R 4 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0197] X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ;

[0198] R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

[0199] In a preferred embodiment of formula (I), wherein R 1 and R 2 are independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, or -CH2OH; preferably, R 1 and R 2 are all selected from -CH3;

[0200] R 3 is selected from -H, -CH3, or -CH2CH3;

[0201] R 4 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0202] X 3 Select N, or CR 7 ;X 4Select N, or CR 8 ;

[0203] R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

[0204] In some embodiments, R 1 and R 2 All are selected from -CD3.

[0205] In some embodiments, R 3 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0206] In some embodiments, R 4 、R 6 Together with the atoms to which they are connected,

[0207] In a preferred embodiment of formula (I), wherein X 3 CR 7 ;X 4 CR 8 ;X 6 N; X 5 and X 7 C; X 1 CH; X 2 CR 6 ;

[0208] R 1 and R 2 are independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH, or -CD3; preferably, R 1 and R 2 are all selected from -CH3, or -CD3;

[0209] R 3 is -CH3, or -CH2CH3;

[0210] R 4 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0211] R 7 and R 8are independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3, preferably, R 7 and R 8 are independently selected from -H or -F.

[0212] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has any of the following structures:

[0213] Ring A and Ring B are each independently selected from phenyl, 5-6 membered heterocycloalkenyl, or C 5-6 cycloalkenyl;

[0214] Among them, R 1 、R 2 、R 3 、R 4 、R 6 、X 3 、X 4 、R 7 and R 8 As defined in any of the above schemes.

[0215] In some embodiments, Ring A and Ring B are each independently selected from phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl, the phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, and 5-6 membered heteroaryl are optionally further substituted by one or more selected from halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxyl C 1-6 Alkyl, and C 1-6 The substituents of the alkoxy group are substituted.

[0216] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has any of the following structures:

[0217] Ring A is independently selected from phenyl, 5-6 membered heterocycloalkenyl, or C 5-6 cycloalkenyl;

[0218] Among them, R 1 、R 2 、R 3 、R 4 、R 6 、X 3 、X 4 、R 7 , and R 8As described in any of the previous options.

[0219] In a preferred embodiment of formula (I), ring A is selected from phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl, the phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, and 5-6 membered heteroaryl are optionally further substituted by one or more selected from halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 The substituent of alkyl-C(O)- is substituted.

[0220] In another aspect, the present disclosure provides a compound represented by the following general formula (II), a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0221] X 1 Select N, or CR 5 ;

[0222] X 3 Select N, or CR 7 ;

[0223] X 4 Select N, or CR 8 ;

[0224] R 1 and R 2 are independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy C 1-4 Alkyl or deuterated C 1-4 alkyl;

[0225] R 3 Selected from -CN, -OH, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclic group, 3-6 membered heterocycloalkenyl, deuterated C 1-4 alkyl, or 5-6 membered heteroaryl; wherein the phenyl, C 3-6 Cycloalkyl, C 3-6Cycloalkenyl, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 3a Substituted, the R 3a Selected from halogen, C 1-4 Alkyl, or C 1-4 alkoxy;

[0226] L is selected from C 2-4 Alkynylidene, C 1-4 Alkylene, or C 2-4 alkenylene;

[0227] R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, and 4-6 membered heterocyclic groups are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH;

[0228] R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, p-methoxybenzyl, C 1-4 Alkyl-C(O)-, C 3-6 Cycloalkyl, or halogenated C 1-4 Alkyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen;

[0229] R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 cycloalkenyl, or 5-6 membered heteroaryl; the phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkenyl, and C 5-6 Cycloalkenyl is optionally further substituted by one or more alkyl radicals selected from halogen, CN, C 1- 6-alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 substituted by an alkyl-C(O)- substituent;

[0230] R 5 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C1-4 Alkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl.

[0231] In a preferred embodiment of formula (II), R 1 、R 2 Each independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH, or -CD3;

[0232] R 3 is -CH3, or -CH2CH3;

[0233] R 4 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0234] R 5 Selected from -H, -CN, halogenated C 1-3 Alkyl or halogen;

[0235] R 7 and R 8 Each independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -B r , -OCH3, or -OCH2CH3.

[0236] In a preferred embodiment of formula (II), X 1 Selected from CR 5 ;

[0237] R 1 and R 2 are all selected from -CH3, or -CD3;

[0238] R 3 is -CH3, or -CH2CH3;

[0239] R 4 、R 6 Together with the atoms to which they are attached, they form the following groups:

[0240] R 5 Selected from -H or -F;

[0241] R 7 and R 8 are independently selected from -H or -F.

[0242] In a preferred embodiment of formula (II), X 1 Selected from CR5 ;X 3 Selected from CR 7 ;X 4 Selected from CR 8 ;

[0243] R 1 and R 2 are independently selected from C 1-4 Alkyl, preferably -CH3;

[0244] R 3 Selected from C 1-4 Alkyl, preferably -CH3;

[0245] R 4 、R 6 Together with the atoms to which it is connected, it forms a 5-6 membered heteroaryl group, preferably a pyridyl group, more preferably

[0246] R 5 is selected from -H or halogen, preferably H or F, more preferably H;

[0247] R 7 、R 8 are each independently selected from -H.

[0248] In a preferred embodiment of formula (II), the compound, its pharmaceutically acceptable salt or its isomer has any of the following structures:

[0249] Ring B is independently selected from phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl, the phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, and 5-6 membered heteroaryl are optionally further substituted by one or more selected from halogen, CN, C 1-6 Alkyl, C 1- 6 haloalkyl, hydroxyl C 1-6 Alkyl, and C 1-6 Substitution of alkoxy groups;

[0250] Among them, R 3 、R 4 、R 5 、R 6 、X 3 、X 4 、R 7 and R 8 As defined in any of the above schemes.

[0251] In another preferred embodiment of formula (I), wherein X 1 Selected from N, X 5 Selected from N, X2 Selected from CR 6 , X 6 Selected from C, X 7 Selected from C.

[0252] In another preferred embodiment of formula (I), wherein X 1 Selected from N, X 2 Selected from CR 6 , X 5 Selected from C, X 6 Selected from C, X 7 Selected from N.

[0253] In a preferred embodiment of formula (I), R 1 and R 2 are independently selected from -CH3, -CH2CH3, or -CH(CH3)2; preferably, R 1 and R 2 are all selected from -CH3;

[0254] R 3 Selected from -CH3;

[0255] R 4 Selected from -H;

[0256] R 6 Selected from -CH3;

[0257] X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ;

[0258] R 7 and R 8 Each independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -OCH3, or -OCH2CH3;

[0259] Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 Cycloalkenyl.

[0260] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has any of the following structures: Among them, R 1 、R 2 、R 3 、R 4 、R 6 、X 3 , and X 4As described in formula (I).

[0261] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-E1):

[0262] Among them, R 3 is H, -CH3 or -CD3;

[0263] R 6 -Br, -Cl, -CH3, -CD3-, (CH3)2CH-, (CH3)2CHCH2-, CF3, CF3CH2-, OH(CH3)2C-, CH3-S(O)2-, cyclopropyl,

[0264] R 4 H; or, R 4 、R 6 Together with the C atom to which it is attached, it forms

[0265] In a preferred embodiment of formula (I-E1), R 3 It is -CH3 or -CD3.

[0266] In a preferred embodiment of formula (I-E1), R 6 It is -Br, -Cl, -CD3, -CF3, CF3CH2- or CH3-S(O)2-.

[0267] In a preferred embodiment of formula (I-E1), R 4 For H.

[0268] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-E2):

[0269] Among them, R 3 is -H, -CH3 or -CD3;

[0270] R 6 -Br, -Cl, -CH3, -CD3-, (CH3)2CH-, (CH3)2CHCH2-, CF3, CF3CH2-, OH(CH3)2C-, CH3-S(O)2-, cyclopropyl,

[0271] R 4 H; or, R 4 、R 6 Together with the C atom to which it is attached, it forms

[0272] In a preferred embodiment of formula (I-E2), R 3 It is H, -CH3 or -CD3.

[0273] In a preferred embodiment of formula (I-E2), R 6 It is -Br, -Cl, -CF3 or cyclopropyl.

[0274] In a preferred embodiment of formula (I-E2), R 4 For H.

[0275] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-F1):

[0276] Among them, R 3 -H, Cl, -CH3, CH3CH2-, -CD3, -CH=CH2 or

[0277] R 6 It is -Cl, -Br, -F, -CH3, CH3CH2-, (CH3)2CH-, -CF3, CH2=C(CH3)-, -CH=CH2, CH3O-, or cyclopropyl;

[0278] R 4 is H;

[0279] Or, R 4 、R 6 Together with the C atom to which it is attached, it forms

[0280] Or, R 3 、R 6 Together with the C atom to which it is attached, it forms

[0281] In a preferred embodiment of formula (I-F1), R 3 It is -H, -CH3, CH3CH2- or -CH=CH2.

[0282] In a preferred embodiment of formula (I-F1), R 6 It is -Cl, -Br, -F, -CH3, CH3CH2-, (CH3)2CH-, -CF3, CH2=C(CH3)-, -CH=CH2, CH3O-, or cyclopropyl.

[0283] In a preferred embodiment of formula (I-F1), R 3 、R 6 Together with the C atom to which it is attached, it forms

[0284] In a preferred embodiment of formula (I-F1), R 4 、R 6 Together with the C atom to which it is attached, it forms

[0285] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-F2):

[0286] Among them, R 3 -H, Cl, -CH3, CH3CH2-, -CD3, -CH=CH2 or

[0287] R 6 It is -Cl, -Br, -F, -CH3, CH3CH2-, (CH3)2CH-, -CF3, CH2=C(CH3)-, -CH=CH2, CH3O-, or cyclopropyl;

[0288] R 4 is H;

[0289] Or, R 4 、R 6 Together with the C atom to which it is attached, it forms

[0290] Or, R 3 、R 6 Together with the C atom to which it is attached, it forms

[0291] In a preferred embodiment of formula (I-F2), R 3 It is -H, -CH3, CH3CH2- or -CH=CH2.

[0292] In a preferred embodiment of formula (I-F2), R 6 It is -Cl, -Br, -F, -CH3, CH3CH2-, (CH3)2CH-, -CF3, CH2=C(CH3)-, -CH=CH2, CH3O-, or cyclopropyl.

[0293] In a preferred embodiment of formula (I-F2), R 3 、R 6Together with the C atom to which it is attached, it forms

[0294] In a preferred embodiment of formula (I-F1), R 4 、R 6 Together with the C atom to which it is attached, it forms

[0295] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-G1):

[0296] Among them, R 3 For CH3-, CH3CH2CH2-, (CH3)2CHCH2-, CH3CH=CH-,

[0297] R 4 It is H, CH3- or cyclopropyl.

[0298] In a preferred embodiment of formula (I-G1), R 3 For CH3CH2CH2-, (CH3)2CHCH2-, CH3CH=CH-,

[0299] In a preferred embodiment of formula (I-G1), R 4 For H.

[0300] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-G2):

[0301] Among them, R 3 CH3-, (CH3)2CHCH2-, CH3CH2CH2-, CH3CH=CH-, or

[0302] R 4 It is H, CH3- or cyclopropyl.

[0303] In a preferred embodiment of formula (I-G2), R 4 For H.

[0304] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-H1):

[0305] R3 is -CH3;

[0306] R 7 H or F;

[0307] R 8 H or F;

[0308] R 4 、R 6 Together with the C atom to which it is attached, it forms

[0309] In a preferred embodiment of formula (I-H1), R 7 H; R 8 For H.

[0310] In a preferred embodiment of formula (I-H1), R 7 H; R 8 For F.

[0311] In a preferred embodiment of formula (I-H1), R 7 F; R 8 For H.

[0312] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer has a structure as shown in formula (I-H2):

[0313] R 3 is -CH3;

[0314] R 7 H or F;

[0315] R 8 H or F;

[0316] R 4 、R 6 Together with the C atom to which it is attached, it forms

[0317] In a preferred embodiment of formula (I-H1), R 7 H; R 8 For H.

[0318] In a preferred embodiment of formula (I-H1), R 7 H; R 8 For F.

[0319] In a preferred embodiment of formula (I-H1), R 7 F; R 8 For H.

[0320] Any substituent and any optional group in the technical solution described in the present disclosure can be combined with each other to form a new complete technical solution. The new technical solution formed has the same or similar technical effects as the solution recorded in the present disclosure and is included in the scope of the present disclosure.

[0321] In a preferred embodiment of formula (I), the compound, its pharmaceutically acceptable salt or its isomer is selected from any one of the following structures:

[0322] In a second aspect, the present disclosure further provides a pharmaceutical preparation composition comprising a compound according to any embodiment of the present disclosure, a pharmaceutically acceptable salt thereof, or an isomer thereof, and one or more pharmaceutically acceptable excipients. The pharmaceutical preparation composition can be in any pharmaceutically acceptable dosage form.

[0323] According to the present disclosure, pharmaceutically acceptable excipients are substances that are non-toxic, compatible with the active ingredient, and otherwise biologically suitable for use in an organism. The selection of a specific excipient will depend on the mode of administration or the type and condition of disease used to treat a particular patient. Examples of pharmaceutically acceptable excipients include, but are not limited to, conventional solvents, diluents, dispersants, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, adhesives, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, release agents, coating agents, flavoring agents, antioxidants, and the like, which are conventional in the pharmaceutical field. If necessary, flavoring agents, preservatives, and sweeteners may also be added to the pharmaceutical formulation composition.

[0324] In a third aspect, the present disclosure further provides a drug containing the compound according to any embodiment of the present disclosure, a pharmaceutically acceptable salt thereof, or an isomer thereof, for preventing and / or treating tumor diseases mediated by PKMYT1.

[0325] In another embodiment, the PKMYT1-mediated tumor disease is a tumor disease caused by CCNE1 overexpression and / or FBXW7 inactivation mutation.

[0326] In another aspect, the present disclosure further provides a drug containing the compound described in any embodiment of the present disclosure, a pharmaceutically acceptable salt thereof, or an isomer thereof, for preventing and / or treating tumor diseases caused by CCNE1 overexpression and / or FBXW7 inactivation mutation.

[0327] In another embodiment, the neoplastic disease is selected from one or more of ovarian cancer, breast cancer, gastric cancer, esophageal cancer, lung cancer, endometrial cancer, and colorectal cancer.

[0328] In a fourth aspect, the present disclosure further provides a method for treating tumor diseases mediated by PKMYT1, comprising administering a therapeutically effective amount of the compound according to any embodiment of the present disclosure, a pharmaceutically acceptable salt thereof, or an isomer thereof to a subject.

[0329] In another embodiment, the PKMYT1-mediated tumor disease is a tumor disease caused by CCNE1 overexpression and / or FBXW7 inactivation mutation.

[0330] In another embodiment, the neoplastic disease is selected from one or more of ovarian cancer, breast cancer, gastric cancer, esophageal cancer, lung cancer, endometrial cancer, and colorectal cancer.

[0331] In another aspect, the present disclosure further provides a method for treating tumor diseases caused by CCNE1 overexpression and / or FBXW7 inactivation mutation, comprising administering a therapeutically effective amount of the compound of any embodiment of the present disclosure, a pharmaceutically acceptable salt thereof, or an isomer thereof to a subject.

[0332] Technical Effects

[0333] The disclosed compounds have good PKMYT1 inhibitory activity, good in vitro and in vivo tumor inhibitory activity, and pharmacokinetic properties; the disclosed compounds also have good physicochemical properties, such as good solubility, which is more conducive to drug absorption and formulation development.

[0334] Description and Definition

[0335] In this disclosure, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of this disclosure, definitions of some terms are provided below. When the definitions of terms provided in this disclosure are inconsistent with the meanings commonly understood by those skilled in the art, the definitions and interpretations of the terms provided in this disclosure shall prevail.

[0336] The "PKMYT1-mediated diseases" mentioned in the present disclosure refer to diseases related to PKMYT1 targets, which can be diseases caused by abnormal expression due to mutation or deletion of PKMYT1 itself, or diseases caused by abnormal expression of PKMYT1 due to abnormalities in other related genes / targets (such as genes / targets that have a synthetic lethal relationship with PKMYT1).

[0337] As used herein, "CCNE1 overexpression" refers to CCNE1 expression levels that are higher than those in normal cells. Compared to normal cells, cells overexpressing CCNE1 exhibit higher CCNE1 activity. For example, normal diploid cells exhibit two copies of CCNE1, while cells overexpressing CCNE1 exhibit at least three copies. CCNE1 overexpression can be measured by identifying the expression level of the gene product in the cell (e.g., CCNE1 mRNA transcript count or CCNE1 protein level).

[0338] The “FBXW7 inactivating mutation” described in the present disclosure includes various types of mutations that inactivate FBXW7 gene expression, including but not limited to base insertion, deletion, mutation, substitution, chemical modification, etc.

[0339] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0340] The "pharmaceutically acceptable salts" described in the present disclosure refer to salts formed by acidic functional groups (such as -COOH, -OH, -SO3H, etc.) present in the compound and suitable inorganic or organic cations (bases), including salts formed with alkali metals or alkaline earth metals, ammonium salts, and salts formed with nitrogen-containing organic bases; and salts formed by basic functional groups (such as -NH2, etc.) present in the compound and suitable inorganic or organic anions (acids), including salts formed with inorganic acids or organic acids (such as carboxylic acids, etc.).

[0341] The terms "therapeutically effective amount" and "effective amount" refer to an amount sufficient to produce a beneficial or desired effect when administered to a subject and within a range that the subject can tolerate; such effects may include preventing the development of tumors, and / or inhibiting tumor growth, and / or limiting tumor spread, and / or reducing tumor volume, and / or ameliorating clinical symptoms or indicators associated with cancer. However, it should be understood that the total daily dosage of the disclosed compounds should be determined by the attending physician within the scope of sound medical judgment.

[0342] The “isomers” described in the present disclosure include geometric isomers and stereoisomers, such as atropisomers, cis-trans isomers, enantiomers, diastereomers, tautomers, and racemic mixtures and other mixtures thereof, all of which fall within the scope of the present disclosure. The term “enantiomer” refers to stereoisomers that are mirror images of each other. The term “tautomer” refers to a type of functional group isomer that has different hydrogen attachment points due to one or more double bond displacements, for example, a ketone and its enol form are keto-enol tautomers. The term “diastereomer” refers to a stereoisomer in which a molecule has two or more chiral centers and is not a mirror image between the molecules. The term “cis-trans isomer” refers to different spatial configurations in which double bonds or single bonds of ring carbon atoms in a molecule cannot rotate freely. The term “atropisomer” refers to stereoisomers that can be separated because single bond rotation is hindered or rotates very slowly.

[0343] Stereoisomers of the compounds disclosed herein can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. For example, one enantiomer of a compound disclosed herein can be prepared by asymmetric catalysis or chiral auxiliary derivatization techniques. Alternatively, a single stereoisomer of the compound can be obtained from a mixture using chiral resolution techniques. Alternatively, the enantiomer can be prepared directly using chiral starting materials. Separation of optically pure compounds disclosed herein is typically accomplished using preparative chromatography, employing chiral chromatographic columns to achieve the purpose of separating chiral compounds.

[0344] The compounds disclosed in the present invention exist in the form of atropisomers, and such structures can be considered as extensions of the chiral center. Looking along the C1-C1' axis, the groups on both sides of the carbon center C1 closer to the observer are ranked first and second in priority; the groups on both sides of C1' at the other end are ranked third and fourth, and then sorted according to the priority of the groups, clockwise for R configuration and counterclockwise for S configuration. It should be noted that the results are the same regardless of which end of the C-C' axis is observed. In the present disclosure, the compound structures are represented by solid line bonds. When , it means that it is located above the plane where most of the atoms in the compound molecule are located, for example Taking the double-fused ring as the plane, the structure represented by the real bond in the benzene ring structure is located above the plane.

[0345] It is known to those skilled in the art that when a cyclic compound has a coplanar delocalized system and the number of π electrons is 4n+2, the ring is aromatic. The expression of the aromatic structure in the compound can be expressed by using dotted lines to represent electron delocalization or by alternating single and double bonds. For example, the structure of a benzene ring can be drawn as Can also be

[0346] The term "optionally substituted" as used herein refers to two situations in which one or more hydrogen atoms of the substituted group may be "substituted" or "unsubstituted" by one or more substituents.

[0347] "PMB" refers to p-methoxybenzyl.

[0348] When the substituent structure appears A truncated bond indicates that the bond is the connecting bond of a substituent, e.g. Indicates that the pyrimidine ring is connected to a given group or a given structural formula through a C atom. A dash "-" appearing in a substituent structure indicates the point of attachment for the substituent, for example, -SCH3 is connected to a given group or a given structural formula through a sulfur atom.

[0349] When a substituent's bond crosses two atoms in a ring, the substituent may be bonded to any atom in the ring. The substituent R can be substituted at any position on the benzene ring.

[0350] When a substituent is listed without indicating the atom via which the substituent is attached to a given group or a given formula, then the substituent may be attached via any bondable atom thereof.

[0351] In the present disclosure, when the structural unit Define R 3 、R 6 When the carbon atoms connected to it form the following ring structure: At this time, the newly formed ring Due to the presence of a shared double bond, they are defined as "heterocycloalkenyl" and "cycloalkenyl". In this disclosure, the "R 2 、R 8 Together with the atoms it is connected to form C 5-6 Cycloalkenyl", "R 3 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl", "R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 This definition applies to "cycloalkenyl".

[0352] The "alkyl" mentioned in the present disclosure refers to a group derived from a branched or straight chain saturated aliphatic alkane with a specified number of carbon atoms by removing one hydrogen atom. For example, "C 1-10 "Alkyl" refers to C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 Alkyl groups, including "C 1-6 alkyl", "C 1-4 Alkyl", "C 1-3 "alkyl"; specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, sec-butyl, 2-methylbutyl, 1,1-dimethylbutyl, etc.

[0353] The "haloalkyl" mentioned in the present disclosure refers to a group obtained by replacing one or more hydrogen atoms in an alkyl group with halogen atoms, such as "fluoromethyl" including monofluoromethyl, difluoromethyl, and trifluoromethyl. Preferably, the "haloalkyl" mentioned in the present disclosure is "haloC 1-6 Alkyl", "halogenated C 1-4 Alkyl". Alkyl is as defined above.

[0354] The "hydroxyalkyl" mentioned in the present disclosure refers to a group derived from an alkyl group in which one or more hydrogen atoms are replaced by a hydroxyl group. The "hydroxyalkyl" mentioned in the present disclosure includes "hydroxy C 1-6 Alkyl", "Hydroxy C 1-4 alkyl"; specific examples include but are not limited to -CH2OH, -CH2CH2OH, -CH(OH)CH3, -CH2CH2CH2OH, wait.

[0355] The "alkoxy" in the present disclosure refers to an alkyl group defined herein connected to another group through an oxygen atom, i.e. "alkyl-O-". 1-6 Alkoxy" (structure is C 1-6 Alkyl-O-), "C 1-4 "alkoxy", specific examples include but are not limited to methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, etc.; preferably, the "alkoxy" described in the present disclosure is C 1-4 Alkoxy, more preferably C 1-3 Alkoxy.

[0356] The "halogenated alkoxy" in the present disclosure refers to a group obtained by replacing one or more hydrogen atoms in an alkoxy group with halogen atoms. Preferably, the "halogenated alkoxy" in the present disclosure is a "halogenated C 1-6 Alkoxy", "halogenated C 1-4 Alkoxy". Specific examples of the present disclosure such as "fluoromethoxy" include monofluoromethoxy, difluoromethoxy, trifluoromethoxy; and also include but are not limited to: -OCH2CF3, -OCHFCH3, etc. Alkoxy is as defined above.

[0357] The "alkenyl" mentioned in the present disclosure refers to a group derived from a straight-chain or branched alkene (containing at least one double bond) by removing a hydrogen atom, including "C 2-6 Alkenyl", "C 2-5Alkenyl", "C 2-4 Alkenyl", "C 2-3 "Alkenyl", specific examples include but are not limited to: -CH=CH2, -CH=CHCH3, -C(CH2)=CH2, -CH=CHCH2CH3, -CH2CH=CHCH3, etc.

[0358] The "alkynyl" mentioned in the present disclosure refers to a group derived from a straight-chain or branched alkyne (containing at least one triple bond) by removing a hydrogen atom, including "C 2-5 Alkynyl", "C 2-4 Alkynyl", "C 2-3 Specific examples include, but are not limited to: -C=CH, -C=CHCH3, CH=CHCH2-, CH=CC=C-, etc.

[0359] The "alkylene" mentioned in the present disclosure refers to a group derived from a branched or straight chain saturated aliphatic alkane by removing two hydrogen atoms, and the removed hydrogen atoms may be derived from the same carbon atom or different carbon atoms; the "alkylene" mentioned in the present disclosure is preferably a "straight chain alkylene"; the "alkylene" includes "C 1-6 Alkylene", "C 1-4 Alkylene", "C 1-2 "Alkylene"; specific examples include, but are not limited to, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH(CH2)CH2-, -CH2CH2CH2CH2-, -CH(CH2)CH2CH2-, -CH(CH2CH2)CH2-, -C(CH2)(CH2)CH2-, -CH2CH2CH2CH2CH2-, etc.

[0360] The "alkenylene" mentioned in the present disclosure refers to a group derived from a straight-chain or branched olefin (containing at least one double bond) by removing two hydrogen atoms. The removed hydrogen atoms may be derived from the same carbon atom or different carbon atoms. The "alkenylene" includes "C 2-6 Alkenylene", "C 2-4 "Alkenylene"; specific examples include, but are not limited to, -CH=CH-, -CH=CHCH2-, -CH=C(CH3)CH2-, -CH=CHCH2CH=CH-, -CH2CH=CHCH2-, -CH2CH=CHCH2CH2-.

[0361] The "alkynylene" mentioned in the present disclosure refers to a group derived from a straight-chain or branched alkyne (containing at least one triple bond) by removing two hydrogen atoms. The removed hydrogen atoms may be derived from the same carbon atom or different carbon atoms. The "alkynylene" includes "C 2-6 Alkynylidene", "C 2-4"Alkynylene"; specific examples include, but are not limited to, -C≡C-, -C≡C-CH2-, -C≡C-CH2CH2CH2-, -CH2-C≡C-CH2CH2CH2-, -C≡C-CH2C≡C-, and -C≡C-CH2CH=CH-.

[0362] As used herein, a "cycloalkyl" refers to a saturated cyclic group derived from a monocyclic cycloalkane by removing a hydrogen atom. In such a cycloalkyl group, except for the carbon atom bonded to a given group or structural formula, the remaining ring carbon atoms may be further oxidized, i.e., to form C(O). Examples of cycloalkyl groups include "3-8 membered cycloalkyl," "3-6 membered cycloalkyl," "3-5 membered cycloalkyl," and "4-6 membered cycloalkyl." Specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0363] The "cycloalkenyl" mentioned in the present disclosure refers to a "cycloalkyl" in which one or more ring bonds are double bonds and the cycloalkenyl is not aromatic. In the cycloalkenyl, except for the carbon atoms bonded to a given group or a given structural formula, other ring carbon atoms may be further oxidized, i.e., to form C(O). The cycloalkenyl includes "3-8 membered cycloalkenyl", "3-6 membered cycloalkenyl", "3-5 membered cycloalkenyl", and "5-6 membered cycloalkenyl". Specific examples include but are not limited to

[0364] The "heterocyclyl" described in the present disclosure refers to a saturated cyclic group derived from the replacement of one or more ring carbon atoms in a cycloalkyl group with a heteroatom. The heteroatom is generally selected from N, O, and S; the carbon atoms or heteroatoms in the heterocyclyl may be further oxidized, i.e., to form C(O), N(O), SO, or SO2; preferably, the heteroatoms are independently selected from 1-3 N and / or O. The heterocyclyl includes "3-8 membered heterocyclyl," "3-6 membered heterocyclyl," "3-5 membered heterocyclyl," "4-6 membered heterocyclyl," and "5-6 membered heterocyclyl." Specific examples include, but are not limited to, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, and morpholinyl.

[0365] The term "heterocycloalkenyl" as used herein refers to a "heterocyclic group" in which one or more of the ring bonds is a double bond and the heterocycle is non-aromatic. Preferably, the heteroatoms are independently selected from 1-3 nitrogen atoms and / or oxygen atoms. Examples of heterocyclic groups include "3-8 membered heterocycloalkenyl," "3-6 membered heterocycloalkenyl," "3-5 membered heterocycloalkenyl," and "5-6 membered heterocycloalkenyl." Specific examples include, but are not limited to: wait.

[0366] The "aryl group" described in the present disclosure refers to a monocyclic or polycyclic group composed of ring carbon atoms and having aromatic properties, and specific examples include but are not limited to: phenyl and naphthyl.

[0367] The "heteroaryl" described in the present disclosure refers to a monocyclic group with aromatic properties in which at least one ring atom is a heteroatom, and the heteroatom is generally selected from N, O, and S. The carbon atoms or heteroatoms in the heterocyclic group may be further oxidized, and the oxidization generally conforms to the rules and conditions for valence bond formation, i.e., forming C(O), N(O), SO, and SO2. Preferably, the heteroatoms are independently selected from 1-3 N and / or O. The heteroaryl group includes "5-6 membered heteroaryl"; specific examples include, but are not limited to, pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl, pyridinyl, and pyrimidinyl.

[0368] Combinations of substituents and / or variables described herein are permitted only if they result in stable compounds or useful synthetic intermediates. Persons skilled in the art will be able to exclude from this disclosure situations that are clearly beyond the ordinary knowledge of the art or unreasonable situations. A stable compound or stable structure is one that is sufficiently stable to withstand chemical reactions, be isolated to a useful degree of purity, and be formulated into an effective therapeutic agent. DETAILED DESCRIPTION

[0369] In the examples disclosed herein, the title compound names were derived from the compound structures using ChemDraw. In the event of inconsistencies between the compound name and the compound structure, the compound structure was determined using a combination of relevant information and reaction routes. If other methods were unavailable for confirmation, the given compound structure was used as the standard.

[0370] The preparation methods of some compounds disclosed herein refer to the preparation methods of the aforementioned similar compounds. Those skilled in the art should be aware that when using or referring to the preparation methods cited, the feed ratio of reactants, reaction solvent, reaction temperature, etc. can be appropriately adjusted according to the different reactants.

[0371] The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples disclosed herein.

[0372] 1. Summary of experimental instruments:

[0373] The structures of the compounds disclosed herein are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Varian 400M or Bruker Ascend 400 NMR instrument in deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), deuterated chloroform (CDCl3), or heavy water (D2O), with tetramethylsilane (TMS) as the internal standard.

[0374] The starting materials in the examples of the present disclosure are known and commercially available, or can be synthesized using or according to methods known in the art.

[0375] 2. Synthesis Example

[0376] The purification methods described in the embodiments of the present disclosure include, but are not limited to, methods known in the art, such as silica gel chromatography, preparative HPLC chromatography, and the like.

[0377] The compounds disclosed herein are mainly synthesized using the following scheme:

[0378] Solution 1

[0379] Compounds of Formula A can be prepared from Intermediate 1 by the following steps: Intermediate 1 is brominated to produce Intermediate 2, which is then hydroxychlorinated to produce Intermediate 3, which is then subjected to a substitution reaction with 3-methoxy-2,6-dimethylaniline in a base and an organic solvent, or a coupling reaction catalyzed by palladium / chiral phosphine ligands to produce Intermediate 4. Intermediate 4 is then cyclized by coupling with malondicyanide to produce Intermediate 5. Intermediate 5 is then treated in an organic solvent with the addition of an acid to produce a compound of Formula A. This compound of Formula A is then subjected to chiral preparation methods to produce two chiral isomers.

[0380] Option 2:

[0381] As shown in Scheme 2, products 7 and 12 can be prepared from intermediate 6 as a starting reactant: intermediate 6 undergoes cyano substitution or bromination reaction to prepare products 7 and 8, and is coupled with an organoboron reagent to prepare intermediate 9; when R 3 When the substituent is an olefin, intermediate 9 is reduced to produce product 10. Intermediate 6 is coupled with an alkyne reagent to produce product 11, and reacted with an amine compound to produce product 12.

[0382] R m 、R P 、R q are independently selected from H, C 1-4 Alkyl, C 3-6 cycloalkyl, or R m With Rq and connected groups to form C 3- 6-membered cycloalkenyl or 3-6-membered heterocycloalkenyl.

[0383] Option 3

[0384] As shown in Scheme 3, 12-15 can be prepared from intermediate 9: intermediate 9 is reacted with an amine to produce intermediate 13, and intermediate 13 is hydrolyzed as described in Scheme 1 to produce product 12. Intermediate 9 is coupled with an organoboron reagent to produce intermediate 14, and intermediate 14 is hydrolyzed to produce product 15.

[0385] Option 4

[0386] As described in Scheme 4, the compound represented by the general formula A can be prepared from the compound represented by the general formula intermediate 16 by the following steps: intermediate 16 is subjected to bromination reaction to obtain intermediate 17, intermediate 17 is subjected to Sandermeyer bromination reaction of the amino group to prepare intermediate 18, and intermediate 18 is subjected to aromatic amination, malondicyanide coupling cyclization, and acid hydrolysis as described in Scheme 1 above to obtain the compound represented by the general formula A.

[0387] Option 5

[0388] As described in Scheme 5, the compound represented by general formula A can be prepared from the general intermediate 19 through aromatic amination, malondicyanide coupling cyclization, and acidolysis as described in Scheme 1 above.

[0389] Option 6

[0390] As described in Scheme 5, the compound represented by general formula A can be prepared from the general intermediate 22 through aromatic amination, malondicyanide coupling cyclization, and acidolysis as described in Scheme 1 above.

[0391] The products and racemic intermediate compounds in the above schemes 2-5 can be prepared into two chiral isomers respectively by the chiral preparation method shown in scheme 1.

[0392] Specific compound synthesis example:

[0393] Example 1

[0394] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0395] In a single-necked flask, 5-bromo-4-chloro-2-methylpyrimidine (9 g, 43.48 mmol), 3-methoxy-2,6-dimethylaniline (7.88 g, 52.19 mmol), and 2,6-lutidine (8.61 g, 86.96 mmol) were dissolved in N-methylpyrrolidone (180 ml) and stirred at 130°C for 40 hours. After completion of the reaction, the reaction mixture was extracted with saturated aqueous ammonium chloride (300 ml) and ethyl acetate (200 ml). After phase separation, the ethyl acetate phase was collected, and the aqueous phase was extracted again with ethyl acetate (100 ml). The combined organic phases were extracted and washed sequentially with saturated ammonium chloride (100 ml x 2), water (100 ml), and saturated brine (100 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was purified to give 5-bromo-N-(3-methoxy-2,6-dimethylphenyl)-2-methylpyrimidin-4-amine (2.3 g). MS (ESI) M / Z: 323.7 [M+H + ].

[0396] Under nitrogen protection, malononitrile (2.34 g, 35.50 mmol) and sodium tert-butoxide (3.40 g, 35.50 mmol) were dissolved in tetrahydrofuran (44 ml) and reacted at room temperature for 30 minutes. 5-Bromo-N-(3-methoxy-2,6-dimethylphenyl)-2-methylpyrimidin-4-amine (2.3 g, 7.10 mmol) and bis[(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (520 mg, 0.71 mmol) were then added thereto, the system was purged with nitrogen again, and the reaction was continued at 100 degrees Celsius for 3 hours. After TLC monitoring showed the disappearance of the starting material, water (60 ml) and ethyl acetate (40 ml) were added and stirred to separate the phases. The organic phase was washed with saturated brine (40 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (1.2 g). MS (ESI) M / Z: 308.0 [M+H + ].

[0397] 6-Amino-7-(3-methoxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (1.2 g, 3.91 mmol) was dissolved in sulfuric acid / water (13 / 1, 6 ml). Methanesulfonic acid (14 g, 144.63 mmol) was added and stirred at room temperature for 2 hours. DL-methionine (2.33 g, 15.64 mmol) was then added to the reaction mixture and stirred at 40°C overnight. The reaction was stopped after TLC monitoring showed the disappearance of the starting material. The pH was adjusted to approximately 7 by dropwise addition of sodium hydroxide / dipotassium hydrogen phosphate aqueous solution to the reaction mixture. The crude product was filtered and purified to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (690 mg). MS (ESI) M / Z: 311.8 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.62 (s, 1H), 8.88 (s, 1H), 7.11-7.03 (m, 3H), 6.94 (d, J=8.3Hz, 1H), 6.85 (s, 2H), 2.44 (s, 3H), 1.75 (s, 3H), 1.67 (s, 3H).C 16 H 17 N5O2.

[0398] Example 2

[0399] S-6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0400] 100 mg of Example 1 compound was obtained by chiral HPLC (29.9 mg, 30% yield). [α] 20 D =+55.54° (c=0.088, MeOH), >99%ee.

[0401] Chiral HPLC analysis conditions were: chiral column: Daicel OZ-3 4.6*100 mm 3 μm, temperature: 40°C, mobile phase: CO2 / IPA [1% NH3 (7 M in MeOH)], flow rate: 3.0 mL / min, back pressure: 2000 psi, detection wavelength: 280 nm, cycle time: 3 min, RT = 1.117 min.

[0402] SFC separation conditions were: equipment: SFC-150mg / m (waters), chiral column: Daicel OZ (25*250mm, 10μm), temperature: 30°C, mobile phase: CO2 / IPA [0.5% NH3 (7M in MeOH)] = 65 / 35, flow rate: 100mL / min, back pressure: 100bar, detection wavelength: 214nm, cycle time: 5.36min, sample solution: 100mg dissolved in 45mL MeOH, injection volume: 2.1ml.

[0403] MS (ESI) M / Z: 312.2 [M+H + ]. 1 H NMR (400MHz, DMSO) δ9.58 (s, 1H), 8.88 (s, 1H), 7.08 (d, J = 8.3Hz, 1H), 7.04 (s, 2 H), 6.93 (d, J=8.3Hz, 1H), 6.83 (s, 2H), 2.44 (s, 3H), 1.75 (s, 3H), 1.67 (s, 3H).

[0404] Example 3

[0405] R-6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0406] 100 mg of the compound of Example 1 was obtained by chiral HPLC (32.1 mg, 32% yield). [α] 20 D =-1.99° (c=0.100, MeOH), 98%ee.

[0407] Chiral HPLC analysis conditions were: chiral column: Daicel OZ-3 4.6*100 mm 3 μm, temperature: 40°C, mobile phase: CO2 / IPA [1% NH3 (7 M in MeOH)], flow rate: 3.0 mL / min, back pressure: 2000 psi, detection wavelength: 280 nm, cycle time: 3 min, RT = 1.511 min.

[0408] SFC separation conditions were: equipment: SFC-150mg / m (waters), chiral column: Daicel OZ (25*250mm, 10μm), temperature: 30°C, mobile phase: CO2 / IPA [0.5% NH3 (7M in MeOH)] = 65 / 35, flow rate: 100mL / min, back pressure: 100bar, detection wavelength: 214nm, cycle time: 5.36min, sample solution: 100mg dissolved in 45mL MeOH, injection volume: 2.1ml.

[0409] MS (ESI) M / Z: 312.2 [M+H + ]. 1 H NMR (400MHz, DMSO) δ9.58 (s, 1H), 8.88 (s, 1H), 7.08 (d, J = 8.3Hz, 1H), 7.04 (s, 2 H), 6.93 (d, J=8.3Hz, 1H), 6.83 (s, 2H), 2.44 (s, 3H), 1.75 (s, 3H), 1.67 (s, 3H).

[0410] Example 4

[0411] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(trifluoromethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0412] 2-(Trifluoromethyl)pyrimidin-5-amine (9.5 g, 58.3 mmol) was dissolved in acetonitrile (100 ml), and N-bromosuccinimide (12.5 g, 70 mmol) was added. The mixture was allowed to react at room temperature overnight. After TLC monitoring showed the disappearance of the starting material, the reaction was stopped and extracted with water (200 ml) and ethyl acetate (200 ml). The organic phase was washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the organic solvent to obtain a crude mixture. The resulting crude product was purified to give 4-bromo-2-(trifluoromethyl)pyrimidin-5-amine (9.2 g). 1 H NMR (400MHz, CDCl3) δ8.19 (s, 1H), 4.56 (br, 2H).C5H3BrF3N3.

[0413] 4-Bromo-2-(trifluoromethyl)pyrimidin-5-amine (4 g, 16.6 mmol) was dissolved in acetonitrile (40 ml). t-Butyl nitrite (2.56 g, 25 mmol) was added in an ice-water bath, followed by cuprous bromide (2.83 g, 19.92 mmol). The mixture was stirred at 60°C for 2 hours. After TLC monitoring showed the disappearance of the starting material, water (100 ml) and ethyl acetate (100 ml) were added to extract the reaction system, and the organic phase was collected. The organic phase was then washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture, which was purified to yield 4,5-dibromo-2-(trifluoromethyl)pyrimidine (700 mg). 1 H NMR (400MHz, CDCl3) δ8.91 (s, 1H).C5HBr2F3N3.

[0414] 4,5-Dibromo-2-(trifluoromethyl)pyrimidine (700 mg, 2.31 mmol), 3-methoxy-2,6-dimethylaniline (384 mg, 2.54 mmol), and 2.6-lutidine (400 mg, 3.74 mmol) were dissolved in N-dimethylpyrrolidone (20 ml), and the mixture was stirred at 95°C overnight. After TLC monitoring showed the disappearance of the starting material, water (30 ml) and ethyl acetate (30 ml) were added and stirred to separate the layers. The organic phase was separated, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 5-bromo-N-(3-methoxy-2,6-dimethylphenyl)-2-(trifluoromethyl)pyrimidin-4-amine (650 mg). MS (ESI) M / Z: 378.2 [M+H + ].

[0415] Malononitrile (585 mg, 8.6 mmol) was dissolved in ethylene glycol dimethyl ether (40 ml). Sodium tert-butoxide (845 mg, 8.6 mmol) was then added to the reaction system. The nitrogen atmosphere was replaced and the reaction was stirred at room temperature for 30 minutes. Then, 5-bromo-N-(3-methoxy-2,6-dimethylphenyl)-2-(trifluoromethyl)pyrimidin-4-amine (650 mg, 1.72 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (130 mg, 0.16 mmol) were added to the reaction system and the reaction was continued at 120°C for 4 hours. After TLC monitoring showed the disappearance of the starting material, water (20 ml) and ethyl acetate (40 ml) were added and stirred to separate the liquids. The organic phase was collected, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2-(trifluoromethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (300 mg). MS (ESI) M / Z: 361.9 [M+H + ].

[0416] 6-Amino-2-chloro-7-(3-methoxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (50 mg, 0.14 mmol) was dissolved in sulfuric acid / water (0.1 ml, sulfuric acid / water = 13 / 1). Methanesulfonic acid (500 mg, 5.14 mmol) was then added to the reaction mixture and stirred at room temperature for 2 hours. DL-methionine (83 mg, 0.55 mmol) was then added to the reaction mixture, and the mixture was stirred at 40°C overnight. After TLC monitoring indicated the disappearance of the starting material, sodium hydroxide / dipotassium hydrogen phosphate aqueous solution (20 ml) was added dropwise to the reaction mixture to adjust the pH to approximately 7. Dichloromethane and methanol were then added to separate the layers. The organic phase was collected, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(trifluoromethyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (10.43 mg). MS (ESI) M / Z: 365.5 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.71 (s, 1H), 9.13 (s, 1H), 7.51 (s, 2H), 7.16-7.09 (m, 3H), 6.98 (d, J = 8.3Hz, 1H), 1.75 (s, 3H), 1.67 (s, 3H).C 16 H 14 F3N5O2.

[0417] Example 5

[0418] 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0419] 5-Bromo-2,4-dichloropyrimidine (2 g, 8.78 mmol) was dissolved in NMP (20 ml), and 3-methoxy-2,6-dimethylaniline (1.5 g, 9.65 mmol) and 2,6-lutidine (1.5 g, 14.22 mmol) were added. The reaction solution was stirred at 95 degrees Celsius overnight. After TLC monitoring showed that the starting material disappeared, the reaction was stopped. Water (50 ml) and ethyl acetate (100 ml) were then added to the cooled reaction solution and stirred for extraction. After separation, the organic phase was collected. The organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain a crude mixture. The resulting mixture was purified to give 5-bromo-2-chloro-N-(3-methoxy-2,6-dimethylphenyl)pyrimidin-4-amine (1.8 g). MS (ESI) M / Z: 342.0 [M+H + ].

[0420] Malononitrile (288.6 mg, 4.37 mmol) was dissolved in ethylene glycol dimethyl ether (7.8 ml), and then sodium tert-butoxide (420.3 mg, 4.37 mmol) was added to the reaction system. After nitrogen replacement, the mixture was stirred at room temperature for 30 minutes. Under a nitrogen atmosphere, 5-bromo-2-chloro-N-(3-methoxy-2,6-dimethylphenyl)pyrimidin-4-amine (300 mg, 0.87 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (55 mg, 0.067 mmol) were added to the reaction system. The reaction solution was stirred at 120 degrees Celsius for 4 hours. After TLC monitoring showed the disappearance of the starting material, water (20 ml) and ethyl acetate (20 ml) were added and stirred to separate the liquids. The collected organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-2-chloro-7-(3-methoxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (120 mg). MS (ESI) M / Z: 328.44 [M+H + ].

[0421] 6-Amino-2-chloro-7-(3-methoxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (120 mg, 0.37 mmol) was dissolved in sulfuric acid / water (13 / 1, 0.3 ml), and methanesulfonic acid (1.3 g, 13.6 mmol) was added. The mixture was stirred at room temperature for 2 hours. DL-methionine (219 mg, 1.47 mmol) was then added to the reaction mixture, and the mixture was stirred at 40°C overnight. After TLC monitoring showed the disappearance of the starting material, a sodium hydroxide / dipotassium hydrogen phosphate aqueous solution was added to the mixture to adjust the pH to approximately 7. Solids precipitated, and the crude mixture was filtered to obtain the product. The resulting crude product was purified to yield 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (50 mg). MS (ESI) M / Z: 331.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.69 (s, 1H), 8.89 (s, 1H), 7.33 (s, 2H), 7.15-6.85 (m, 4H), 1.76 (s, 3H), 1.68 (s, 3H).C 15 H 14 ClN5O2.

[0422] Example 6

[0423] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-vinyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0424] Step A: 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (200 mg, 0.6 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (279.2 mg, 1.8 mmol), potassium carbonate (250.2 mg, 1.8 mmol) and tetrakis(triphenylphosphine)palladium (69.8 mg, 0.06 mmol) were dissolved in N,N-dimethylformamide / water (5 / 1, 7.2 ml) and stirred at 4°C under nitrogen atmosphere. The reaction was carried out at 100 degrees Celsius overnight. After LCMS monitoring showed that the starting material disappeared, it was cooled to room temperature. Water (20 ml) and ethyl acetate (20 ml) were extracted and post-processed. The collected organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-vinyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (83.84 mg). MS (ESI) M / Z: 324.5 [M+H + ].1 H NMR (400MHz, DMSO-d6) δ9.62 (s, 1H), 8.97 (s, 1H), 7.19 (s, 2H), 7.09 (d, J=8.3Hz, 1H), 6.98-6.90 (m, 3H), 6.63 ( dd, J=17.2, 10.6Hz, 1H), 6.18 (dd, J=17.2, 2.3Hz, 1H), 5.40 (dd, J=10.5, 2.2Hz, 1H), 1.77 (s, 3H), 1.68 (s, 3H).C 17 H 17 N5O2.

[0425] Example 7

[0426] 6-amino-2-bromo-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0427] 6-Amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (100 mg, 0.3 mmol) was dissolved in 33% hydrobromic acid / acetic acid solution (1 ml), heated to 80 degrees Celsius, and reacted overnight. After LCMS monitoring showed the disappearance of the starting material, the reaction solution was cooled to room temperature. Saturated aqueous sodium bicarbonate solution (20 ml) and ethyl acetate (20 ml) were added and stirred to separate the liquids. The organic phase was collected, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The resulting crude product was purified to give 6-amino-2-bromo-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (45.58 mg). MS (ESI) M / Z: 376.0 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.70 (s, 1H), 8.85 (s, 1H), 7.33 (s, 2H), 7.11 (d, J=8.3Hz, 1H), 7.02-6.93 (m, 3H), 1.76 (s, 3H), 1.68 (s, 3H).C 15 H 14 BrN5O2.

[0428] Example 8

[0429] 6-amino-2-ethyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0430] 6-Amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-vinyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (80 mg, 0.25 mmol) was dissolved in methanol (10 ml), 10% palladium on carbon (100 mg) was added, and hydrogen was introduced into the mixture under normal pressure for 3 hours. After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered and the filtrate was concentrated to obtain a crude product mixture. The crude product was purified to give 6-amino-2-ethyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (17.13 mg). MS (ESI) M / Z: 326.4 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.61 (s, 1H), 8.90 (s, 1H), 7.12-7.03 (m, 3H), 6.93 (d, J=8.2Hz, 1 H), 6.86 (s, 2H), 2.69 (q, J=7.3Hz, 2H), 1.76 (s, 3H), 1.67 (s, 3H), 1.15 (t, J=7.6Hz, 3H).C 17 H 19 N5O2.

[0431] Example 9

[0432] 6-amino-2-(cyclohexyl-1-en-1-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0433] As in Example 6, 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (100 mg, 0.30 mmol) and 2-(cyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (115 mg, 0.91 mmol) were reacted to obtain 6-amino-2-(cyclohexyl-1-en-1-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (19.8 mg). MS (ESI) M / Z: 377.5 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.59 (s, 1H), 8.92 (s, 1H), 7.16-7.05 (m, 3H), 6.93 (m, 1H), 6.87 (m, 2H) 6 .82-6.75(m,1H),2.39(m,2H),2.18-2.05(m,2H),1.77(s,3H),1.69(s,3H),1.66-1.51(m,4H).C21 H 23 N5O2.

[0434] Example 10

[0435] 6-amino-2-cyclohexyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0436] 6-Amino-2-(cyclohexyl-1-en-1-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (15 mg, 0.04 mmol) was dissolved in methanol (2 ml), 10% Pd / C (3 mg) was added, hydrogen was introduced at normal pressure, and the reaction was carried out at room temperature for 1.5 hours. After LCMS monitoring showed the disappearance of the starting material, the filtrate was filtered and collected, and the filtrate was concentrated to give 6-amino-2-cyclohexyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (13.89 mg). MS (ESI) M / Z: 380.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.73 (s, 1H), 8.89 (s, 1H), 7.12-7.01 (m, 3H), 6.95 (d, J=8.3Hz, 1H), 6.84 (s, 2H ), 2.59 (tt, J=11.6, 3.5Hz, 1H), 1.85-1.57 (m, 10H), 1.47 (qt, J=12.5, 3.0Hz, 2H), 1.34-1.12 (m, 4H).C 21 H 25 N5O2.

[0437] Example 11

[0438] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-phenyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0439] 6-Amino-2-chloro-7-(3-methoxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (50 mg, 0.15 mmol), phenylboronic acid (37.3 mg, 0.31 mmol), potassium carbonate (63.3 mg, 0.46 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (7.8 mg, 0.012 mmol) were dissolved in dioxane / water (5 / 1, 3 ml) and reacted at 120°C under a nitrogen atmosphere for 4.5 hours. LCMS monitoring showed that the raw material was completely reacted and then cooled to room temperature. Water (20 ml) and ethyl acetate (20 ml) were added to the reaction system, stirred, extracted and separated. The collected organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2-phenyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (50 mg). MS (ESI) M / Z: 370.3 [M+H + ].

[0440] 6-Amino-7-(3-methoxy-2,6-dimethylphenyl)-2-phenyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (50 mg, 0.14 mmol) was dissolved in sulfuric acid / water (13 / 1, 0.5 ml). Methanesulfonic acid (483.2 mg, 5.03 mmol) was then added to the reaction mixture, and the mixture was stirred at room temperature for 2 hours. DL-methionine (81.1 mg, 0.54 mmol) was then added to the reaction mixture, and the reaction mixture was stirred at 40°C overnight. After TLC monitoring indicated that the starting material had reacted completely, sodium hydroxide / dipotassium hydrogen phosphate aqueous solution was added to the reaction mixture to adjust the pH to approximately 7. The crude mixture was filtered to obtain the product. The resulting crude product was purified to obtain 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-phenyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (5.82 mg). MS (ESI) M / Z: 373.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.63 (s, 1H), 9.09 (s, 1H), 8.17-8.10 (m, 2H), 7.45-7.32 (m, 3 H), 7.23 (s, 2H), 7.13 (d, J=8.3Hz, 1H), 7.01-6.93 (m, 3H), 1.82 (s, 3H), 1.73 (s, 3H).C 21 H 19 N5O2.

[0441] Example 12

[0442] 6-amino-2-(cyclopropylethynyl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0443] 6-Amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (100 mg, 0.30 mmol), cyclopropylacetylene (100 mg, 1.52 mmol), bisacetonitrile palladium chloride (10 mg, 0.04 mmol), cesium carbonate (165 mg, 0.51 mmol) and 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl (28 mg, 0.07 mmol) were dissolved in acetonitrile (4 ml) and stirred at 90 degrees Celsius under a nitrogen atmosphere overnight. After TLC monitoring showed that the raw materials had reacted completely, the reaction system was cooled to room temperature. Water (20 ml) and ethyl acetate (20 ml) were then added to the reaction system and stirred to extract the liquid. The collected organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The resulting crude product was purified to give 6-amino-2-(cyclopropylethynyl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (10.4 mg).

[0444] MS (ESI) M / Z: 362.3 [M+H + ]. 1 HNMR (400MHz, DMSO-d6) δ9.66 (s, 1H), 8.91 (s, 1H), 7.29 (s, 2H), 7.10 (d, J=8.3Hz, 1H), 6.99-6.92 (m, 3H) , 1.74 (s, 3H), 1.65 (s, 3H), 1.50 (tt, J=8.2, 5.0Hz, 1H), 0.90-0.81 (m, 2H), 0.74 (dt, J=4.9, 3.1Hz, 2H).C 20 H 19 N5O2.

[0445] Example 13

[0446] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(piperidin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0447] 6-Amino-2-chloro-7-(3-methoxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (100 mg, 0.31 mmol) was dissolved in n-butanol (2 ml), followed by the addition of piperidine (1 ml) and N,N-diisopropylethylamine (118.6 mg, 0.93 mmol) to the reaction system. The reaction solution was stirred at 150 degrees Celsius for 8 hours. After TLC monitoring showed that the raw material reaction was complete, it was cooled to room temperature. Water (20 ml) and ethyl acetate (20 ml) were added to the reaction system for extraction. After stirring and separating the liquids, the organic phase was collected and washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2-(piperidin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (90 mg). MS (ESI) M / Z: 377.4 [M+H + ].

[0448] 6-Amino-7-(3-methoxy-2,6-dimethylphenyl)-2-(piperidin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (70 mg, 0.19 mmol) was dissolved in sulfuric acid / water (13 / 1, 0.6 ml). Methanesulfonic acid (662 mg, 6.9 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 2 hours, and DL-methionine (111.1 mg, 0.74 mmol) was added to the reaction mixture. The mixture was stirred at 40°C overnight. LCMS monitoring indicated that the reaction of the starting material was complete. A sodium hydroxide / dipotassium hydrogen phosphate aqueous solution was added to the reaction mixture to adjust the pH to approximately 7. The crude product was purified to yield 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(piperidin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (18.9 mg). MS (ESI) M / Z: 380.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.53 (s, 1H), 8.60 (s, 1H), 7.06 (d, J = 8.2Hz, 1H), 6.90 (d, J = 8.2Hz, 1H), 6.7 3(s, 2H), 6.66(s, 2H), 3.49(t, J=5.4Hz, 4H), 1.80(s, 3H), 1.71(s, 3H), 1.52(m, 2H), 1.42(m, 4H).C 20 H 24 N6O2.

[0449] Example 14

[0450] 6-amino-2-cyano-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0451] 6-Amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (90 mg, 0.27 mmol), zinc cyanide (25 mg, 0.21 mmol), 1,1′-bis(diphenylphosphino)ferrocene (12 mg, 0.02 mmol), zinc powder (6 mg, 0.09 mmol), and tris(dibenzylideneacetone)dipalladium (6 mg, 0.006 mmol) were dissolved in dimethylacetamide (3.5 ml) and stirred at 150°C under a nitrogen atmosphere for 3 hours. TLC monitoring indicated complete reaction of the starting material, and the mixture was cooled to room temperature. Water (20 ml) and ethyl acetate (20 ml) were added to the reaction system for extraction. After stirring and separation, the organic phase was collected and washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-2-cyano-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (25.55 mg). MS (ESI) M / Z: 322.4 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.72 (s, 1H), 9.10 (s, 1H), 7.69 (d, J=6.0Hz, 2H), 7.17 (s , 2H), 7.12 (d, J=8Hz, 1H), 7.03-6.95 (d, J=8Hz, 1H), 1.75 (s, 3H), 1.67 (s, 3H).C 16 H 14 N6O2.

[0452] Example 15

[0453] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-((4-methoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0454] 6-Amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (100 mg, 0.3 mmol) was dissolved in n-butanol (2 ml), and then 4-methoxybenzylamine (1 ml) and N,N-diisopropylethylamine (117.1 mg, 0.91 mmol) were added to the reaction solution and stirred at 150 degrees Celsius for 8 hours. After TLC monitoring showed that the raw material reaction was complete, it was cooled to room temperature. Water (20 ml) and ethyl acetate (20 ml) were added for extraction and separation. The collected organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-((4-methoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (64.44 mg). MS (ESI) M / Z: 432.19 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.54 (s, 1H), 8.51 (s, 1H), 7.21-7.02 (m, 4H), 6.96-6.83 (m, 1H), 6.80- 6.70(m, 2H), 6.69-6.58(m, 4H), 4.22(d, J=5.8Hz, 2H), 3.71(s, 3H), 1.78(s, 3H), 1.69(s, 3H).C 23 H 24 N6O3.

[0455] Example 16

[0456] 6-amino-2-(3,6-dihydro-2H-pyran-4-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0457] As in Example 6, 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (120 mg, 0.36 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (240 mg, 1.14 mmol) were reacted to give 6-amino-2-(3,6-dihydro-2H-pyran-4-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (25.21 mg). MS (ESI) M / Z: 379.2 [M+H + ]. 1H NMR (400MHz, DMSO-d6) δ9.59 (s, 1H), 8.96 (s, 1H), 7.16 (d, J = 9.4Hz, 2H), 7.09 (d, J = 8.3Hz, 1H), 6.97-6 .88(m, 3H), 6.78-6.71(m, 1H), 4.20(q, J=2.8Hz, 2H), 3.74(t, J=5.4Hz, 2H), 1.78(s, 3H), 1.69(s, 3H).C 20 H 21 N5O3.

[0458] Example 17

[0459] 6-amino-2-(diethylamino)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0460] As in Example 15, 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (100 mg, 0.3 mmol) was reacted with diethylamine (1 ml) to give 6-amino-2-(diethylamino)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (9.29 mg). MS (ESI) M / Z: 369.0 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.50 (s, 1H), 8.58 (s, 1H), 7.05 (d, J = 8.3Hz, 1H), 6.89 (d, J = 8.2Hz, 1H), 6.68(s, 2H), 6.62(s, 2H), 3.45-3.35(m, 4H), 1.80(s, 3H), 1.72(s, 3H), 0.99(t, J=6.9Hz, 6H).C 19 H 24 N6O2.

[0461] Example 18

[0462] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0463] 6-Amino-2-(3,6-dihydro-2H-pyran-4-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (19 mg, 0.05 mmol) was dissolved in methanol (2 ml), 10% Pd / C (10 mg) was added, hydrogen was introduced at normal pressure, and the reaction was carried out at room temperature for 1.5 hours. After LCMS monitoring showed the disappearance of the starting material, the filtrate was filtered and collected, and the filtrate was concentrated to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (7.03 mg). MS (ESI) M / Z: 382.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.80 (s, 1H), 8.98 (s, 1H), 7.20-7.10 (m, 3H), 7.11 (d, J=8.4Hz, 1H), 6.9 2 (s, 2H), 3.92 (dt, J=11.1, 3.3Hz, 2H), 2.93 (ddd, J=15.4, 9.0, 7.0Hz, 1H), 1.84-1.69 (m, 10H).C 20 H 23 N5O3.

[0464] Example 19

[0465] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0466]

[0467] 2,6-Dimethylpyrimidin-4-ol (2.0 g, 16.11 mmol) was dissolved in chloroform (20 ml), and liquid bromine (3.2 ml, 64.5 mmol) was added dropwise to the reaction mixture under ice-cooling. After the addition was complete, the reaction system was heated to 80°C and stirred overnight. After TLC monitoring showed the disappearance of the starting material, the solvent was removed under reduced pressure, ethyl acetate (50 ml) was added, and the mixture was concentrated under reduced pressure again, and this was repeated three times. The crude mixture was adjusted to pH 7-8 with saturated NaHCO₃. The mixture was extracted with ethyl acetate (80 ml x 3) and dichloromethane / isopropanol (7 / 3, 50 ml x 3). The organic phases were combined, washed with saturated brine (50 ml x 3 times), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude 5-bromo-2,6-dimethylpyrimidin-4-ol (1.316 g). MS (ESI) M / Z: 205.1 [M+H + ]. 1H NMR (400MHz, DMSO) δ12.76 (s, 1H), 2.33 (s, 3H), 2.24 (s, 3H).

[0468] In a single-necked flask, a solution of 5-bromo-2,6-dimethylpyrimidin-4-ol (1.316 g, 6.48 mmol) in phosphorus oxychloride (20 ml) was added, and the reaction system was stirred at 110 degrees Celsius overnight. After TLC monitoring showed the disappearance of the starting material, the solvent was removed under reduced pressure. Ethyl acetate (50 ml) was then added to dissolve the mixture, and the pH was adjusted to 8-9 with saturated NaHCO3 under ice bath. The mixture was extracted with ethyl acetate (80 ml x 3), and the organic phases were separated and combined, washed with saturated brine (50 ml x 3 times), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified to give 5-bromo-4-chloro-2,6-dimethylpyrimidine (1.096 g). MS (ESI) M / Z: 223.0 [M+H + ]. 1 H NMR (400MHz, DMSO) δ2.60 (s, 3H), 2.54 (s, 3H).

[0469] To a 20 mL sealed tube, a solution of 5-bromo-4-chloro-2,6-dimethylpyrimidine (1.096 g, 4.9 mmol) in N-methylpyrrolidone (7 mL) was added. 3-Methoxy-2,6-dimethylaniline (0.9 g, 5.9 mmol) and 2,6-lutidine (1.06 g, 9.9 mmol) were slowly added under a nitrogen atmosphere. The mixture was microwaved at 130°C for 54 hours. LCMS confirmed the formation of the product. After extraction with ethyl acetate (50 mL x 3), the organic phases were combined, washed with saturated brine (20 mL x 3 times), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The resulting mixture was purified to yield 5-bromo-N-(3-methoxy-2,6-dimethylphenyl)-2,6-dimethylpyrimidin-4-amine (883 mg). MS (ESI) M / Z: 336.2 [M+H + ].

[0470] Under a nitrogen stream, sodium tert-butoxide (71 mg, 0.744 mmol), ethylene glycol dimethyl ether (1 ml), and malondicyanide (49 mg, 0.744 mmol) were added to a 10 ml sealed tube. The container was sealed and the reaction mixture was stirred at room temperature for 30 minutes. Then, under a nitrogen stream, 5-bromo-N-(3-methoxy-2,6-dimethylphenyl)-2,6-dimethylpyrimidin-4-amine (50 mg, 0.148 mmol) and PdCl2(dppf)2 (10 mg, 0.015 mmol) were added to the sealed tube. The reaction tube was sealed and heated to 100 degrees Celsius in an oil bath and stirred overnight. TLC analysis showed the formation of new spots, and the reaction mixture was poured into ice water. The organic phases were extracted with ethyl acetate (10 ml x 3) and combined. The organic phases were then washed with saturated brine (5 ml x 3 times), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (30 mg). MS (ESI) M / Z: 322.2 [M+H + ].

[0471] 6-Amino-7-(3-methoxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (30 mg, 0.09 mmol) was dissolved in concentrated sulfuric acid (1 ml) at room temperature. The reaction solution was stirred at room temperature for 5 hours. After LCMS monitoring showed the disappearance of the starting material, crushed ice was added to the reaction solution to quench it, and then the pH value of the reaction solution was adjusted to 9 with concentrated aqueous ammonia. The mixed solution was extracted with ethyl acetate (5 ml × 3) and the organic phases were combined. The organic phases were then washed with saturated brine (5 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude mixture. The resulting crude product was purified to give 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (10 mg). MS (ESI) M / Z: 340.4 [M+H + ].

[0472] In a 50 ml three-necked flask, 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (10 mg, 0.029 mmol) and anhydrous dichloromethane (0.5 ml) were added. A dichloromethane solution of BBr3 (1 M in dichloromethane, 1 ml) was added dropwise to the reaction system under ice-cooling and stirred at room temperature for 1 hour. After TLC monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water, the pH was adjusted to 9 with concentrated ammonia water, and then concentrated under reduced pressure to obtain a crude mixture. The crude product was purified to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (4 mg). MS (ESI) M / Z: 326.2 [M+H + ].1H NMR (400MHz, DMSO) δ9.55 (s, 1H), 7.06 (d, J = 8.3Hz, 1H), 6.92 (d, J = 8.3Hz, 1H), 6.85(s, 2H), 6.47(s, 2H), 2.70(s, 3H), 2.38(s, 3H), 1.75(s, 3H), 1.66(s, 3H).

[0473] Example 20

[0474] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0475] 3-Fluoro-5-methoxyaniline (8.0 g, 56.68 mmol) was dissolved in N,N-dimethylformamide (160 mL). N-chlorosuccinimide (7.57 g, 56.68 mmol) was added to the reaction mixture at room temperature. The reaction system was stirred at room temperature overnight. After TLC monitoring showed the disappearance of the starting material, the reaction mixture was poured into 600 ml of water. Extraction was performed with ethyl acetate (300 mL x 3). The organic phase was washed with saturated brine (100 mL x 3 times), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to obtain 4-chloro-3-fluoro-5-methoxyaniline (6.025 g, yield 60.74%). 1 H NMR (400MHz, DMSO) δ6.19-6.13 (m, 1H), 6.09 (dd, J=11.6, 2.3Hz, 1H), 5.58 (s, 2H), 3.76 (s, 3H).

[0476] To a single-necked flask, a solution of 4-chloro-3-fluoro-5-methoxyaniline (6.0 g, 34.3 mmol) in acetonitrile (170 mL) was added. N-bromosuccinimide (24.4 g, 137.1 mmol) was added to the reaction mixture at room temperature, and the reaction system was stirred overnight at room temperature. After TLC monitoring showed the disappearance of the starting material, the solvent was removed under reduced pressure. The resulting mixture was purified by silica gel column chromatography to yield 2,6-dibromo-4-chloro-3-fluoro-5-methoxyaniline (9.783 g, 85.56% yield). 1 H NMR (400MHz, DMSO) δ5.94 (s, 2H), 3.32 (s, 3H).

[0477] 2,6-Dibromo-4-chloro-3-fluoro-5-methoxyaniline (4.5 g, 13.5 mmol) was dissolved in 1,4-dioxane / water (60 mL / 6 ml). Potassium carbonate (6.53 g, 47.24 mmol), methylboric acid (4.04 g, 67.5 mmol), and PdCl2(dppf) (493 mg, 0.67 mmol) were slowly added under a nitrogen atmosphere. The reaction system was stirred at 100°C overnight. After TLC monitoring showed the disappearance of the starting material, the solvent was removed under reduced pressure, and 50 mL of purified water was added to the system. The mixture was extracted with ethyl acetate (150 mL x 3). The organic phases were combined, washed three times with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to yield 4-chloro-3-fluoro-5-methoxy-2,6-dimethylaniline (1.919 g, yellow liquid, yield 69.80%).

[0478] To a single-necked flask, add a solution of 4-chloro-3-fluoro-5-methoxy-2,6-dimethylaniline (2.0 g, 9.82 mmol) in methanol (80 mL). Under a nitrogen atmosphere, slowly add ammonium formate (12.4 g, 196.43 mmol) and Pd / C (2.0 g, 18.8 mmol). Stir the reaction system at 65°C overnight. After TLC monitoring showed the disappearance of the starting material, filter the mixture and concentrate under reduced pressure. The resulting residue was purified by silica gel column chromatography to afford 3-fluoro-5-methoxy-2,6-dimethylaniline (1.417 g, 85.38% yield). 1 H NMR (400MHz, DMSO) δ6.07 (d, J=12.1Hz, 1H), 4.84 (s, 2H), 3.67 (s, 3H), 1.95-1.82 (m, 6H). MS (ESI) M / Z: 170.0 [M+H + ].

[0479] 3-Fluoro-5-methoxy-2,6-dimethylaniline (707 mg, 4.18 mmol) was dissolved in ethylene glycol dimethyl ether (25 mL) at room temperature. 2,3-Dibromo-5,6-dimethylpyridine (1.0 g, 3.8 mmol), cesium carbonate (3.095 g, 9.5 mmol), Pd2dba3 (348 mg, 0.38 mmol), and XantPhos (440 mg, 0.76 mmol) were added at room temperature. The reaction system was stirred at 100°C overnight. After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with purified water. The mixture was extracted with ethyl acetate (150 mL x 3). The combined organic phases were washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 3-bromo-N-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine (1.166 g, yield 91.9%). 1 H NMR (400MHz, DMSO) δ7.60 (s, 1H), 7.41 (s, 1H), 6.77 (d, J = 11.8Hz, 1H), 3.78 (s, 3H), 2 .07(s, 3H), 2.05(s, 3H), 1.92(d, J=2.0Hz, 3H), 1.90(s, 3H).MS(ESI)M / Z: 355.0[M+H + ].

[0480] To a 20 mL sealed tube, sodium tert-butoxide (1.578 g, 16.42 mmol) was added. Ethylene glycol dimethyl ether (3 mL) was then added dropwise at room temperature. After the addition was complete, the container was sealed and the reaction mixture was stirred at room temperature for 30 minutes. Under a nitrogen flow, 3-bromo-N-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine (1.16 g, 3.28 mmol) and PdCl2(dppf) (240 mg) were added to the sealed tube. The reaction tube was sealed, heated to 100°C in an oil bath, and stirred overnight. TLC analysis showed the formation of new spots, and the reaction mixture was poured into pure water. The reaction mixture was extracted with ethyl acetate (150 mL x 3). The organic phases were combined, washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to give 2-amino-1-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (950 mg, yield 85.6%). 1H NMR (400MHz, DMSO) δ7.40 (s, 1H), 7.10 (d, J=11.7Hz, 1H), 6.92 (s, 2H), 3.86 (s, 3H), 2 .25(s, 3H), 2.25(s, 3H), 1.69(d, J=1.7Hz, 3H), 1.64(s, 3H).MS(ESI)M / Z: 339.1[M+H + ].

[0481] 2-Amino-1-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (950 mg, 2.8 mmol) was dissolved in concentrated sulfuric acid (8 mL) at 0°C. The reaction mixture was stirred at room temperature for 3 hours. After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding crushed ice and the pH was adjusted to 7-8 with saturated aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 2-amino-1-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (732 mg, yield 73.65%). 1 H NMR (400MHz, DMSO) δ7.85 (s, 1H), 7.09 (d, J = 11.8Hz, 1H), 6.87 (s, 2H), 6.67 (s, 2H), 3.86 (s, 3H), 2.26 (s, 3H), 2.24 (s, 3H), 1.69 (d, J=1.5Hz, 3H), 1.65 (s, 3H).MS (ESI) M / Z: 357.2[M+H] + .

[0482] In a 50 mL three-necked flask, 2-amino-1-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (732 mg, 2.05 mmol) and anhydrous dichloromethane (20 mL) were added dropwise. A dichloromethane solution of BBr3 (2N, 10.3 mL, 20.5 mmol) was added dropwise under an ice bath, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitoring showed the disappearance of the starting material. The reaction mixture was quenched with methanol, the pH was adjusted to 8 with concentrated ammonia, and concentrated under reduced pressure to obtain a crude product. The resulting residue was purified by preparative HPLC to obtain the final product, 2-amino-1-(3-fluoro-5-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (4 mg, 71.3% yield). 1H NMR (400MHz, DMSO) δ9.95 (s, 1H), 7.84 (s, 1H), 6.85 (s, 2H), 6.79 (d, J = 11.1Hz, 1H), 6.66 (s, 2H), 2.26(s, 3H), 2.25(s, 3H), 1.65(d, J=1.6Hz, 3H), 1.62(s, 3H).MS(ESI)M / Z: 343.3[M+H] + .

[0483] Example 21

[0484] S-6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0485] 100 mg of the compound of Example 20 was prepared by chiral HPLC. SFC separation method: Instrument: SFC-150 mg / m (waters), chiral column: Daicel-OZ (25*250 mm, 10 μm), temperature: 30°C, mobile phase: CO2 / IPA [0.5% NH3 (7N in MeOH)] = 60 / 40, flow rate: 100 ml / min, back pressure: 100 bar, detection wavelength: 214 nm, cycle time: 8.68 min, RT = 1.643 min. [α] 20 D =+66.47° (c=0.10065, MeOH), 100%ee. 1 H NMR (400MHz, DMSO) δ9.97 (s, 1H), 7.84 (s, 1H), 6.84 (s, 2H), 6.79 (d, J = 11.1Hz, 1H), 6.66 (s, 2H), 2.26(s, 3H), 2.25(s, 3H), 1.65(d, J=1.6Hz, 3H), 1.62(s, 3H).MS(ESI)M / Z: 343.3[M+H] + .

[0486] Example 22

[0487] R-6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2,4-dimethyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0488] 100 mg of the compound of Example 20 was prepared by chiral HPLC. The SFC method was the same as that of Example 21, RT = 2.472 min. 20 D=-6.42° (c=0.10275, MeOH), 96%ee. 1 H NMR (400MHz, DMSO) δ9.95 (s, 1H), 7.84 (s, 1H), 6.85 (s, 2H), 6.79 (d, J = 11.1Hz, 1H), 6.66 (s, 2H), 2.26(s, 3H), 2.25(s, 3H), 1.65(d, J=1.6Hz, 3H), 1.62(s, 3H).MS(ESI)M / Z: 343.3[M+H] + .

[0489] Example 23

[0490] 2-amino-1-(4-fluoro-3-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0491] 4-Fluoro-3-methoxyaniline (1 g, 7.09 mmol) was dissolved in dichloromethane (5 ml) and methanol (5 ml). Bromine (0.96 ml) in dichloromethane (5 ml) and methanol (5 ml) was added dropwise to the reaction mixture, and the mixture was stirred at room temperature for 4 hours. After LCMS monitoring indicated the disappearance of the starting material, aqueous sodium thiosulfate (20 ml) and aqueous sodium carbonate (20 ml) were added, and the mixture was stirred for 10 minutes. Extraction was then performed twice with ethyl acetate (40 ml). The organic phases were combined, washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. Finally, the residue was concentrated under reduced pressure and purified by silica gel column chromatography to yield 2,6-dibromo-4-fluoro-3-methoxyaniline (1.9 g). 1 H NMR (400MHz, CDCl3) δ7.24 (d, J=10.4Hz, 1H), 4.31 (br, 2H), 3.94 (d, J=1.6Hz, 3H). C7H6Br2FNO.MS (ESI) M / Z: 299.9[M+H + ].

[0492] 2,6-Dibromo-4-fluoro-3-methoxyaniline (500 mg, 1.67 mmol) was dissolved in dioxane / water (10 / 1, 5.5 ml). Methylboric acid (301 mg, 5.02 mmol), potassium carbonate (692.3 mg, 5.02 mmol), and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (61.3 mg, 0.08 mmol) were added. The atmosphere was purged with nitrogen three times, and the reaction mixture was stirred at 100°C overnight. After LCMS monitoring indicated the disappearance of the starting material, water (30 ml) was added and the mixture was extracted twice with ethyl acetate (20 ml). The combined organic phases were washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 4-fluoro-3-methoxy-2,6-dimethylaniline (600 mg). 1 H NMR (400MHz, CDCl3) δ6.70 (d, J=12Hz, 1H), 3.83 (s, 3H), 3.47 (br, 2H), 2.12 (d, J=4.4Hz, 6H).C9H 12 FNO.MS (ESI) M / Z: 169.7 [M+H + ].

[0493] 2,3-Dibromo-5,6-dimethylpyridine (200 mg, 0.76 mmol), 4-fluoro-3-methoxy-2,6-dimethylaniline (135.5 mg, 0.8 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (39.7 mg, 0.069 mmol), cesium carbonate (622.1 mg, 1.91 mmol) and trisdibenzylideneacetone dipalladium (35 mg, 0.038 mmol) were added to ethylene glycol dimethyl ether (5 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 90°C overnight. After LCMS monitoring showed the disappearance of the starting material, water (30 ml) was added and extracted twice with ethyl acetate (20 ml). The combined organic phases were washed with saturated brine (10 ml) and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure and purified by silica gel column chromatography to give 3-bromo-N-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine (165 mg). MS (ESI) M / Z: 353.2 [M+H + ].

[0494] Malononitrile (92.8 mg, 1.41 mmol) was dissolved in dioxane (2 ml), sodium tert-butoxide (225.2 mg, 2.34 mmol) was added, the atmosphere was replaced with nitrogen, and the reaction was carried out at room temperature for 30 minutes. 3-Bromo-N-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine (165 mg, 0.47 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (34 mg, 0.047 mmol) were added, the atmosphere was replaced with nitrogen, and the reaction was carried out at 120°C for 2.5 hours. After LCMS monitoring showed the disappearance of the starting material, water (10 ml) was added and extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2-amino-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (100 mg). MS (ESI) M / Z: 339.2 [M+H + ].

[0495] 2-Amino-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (100 mg, 0.3 mmol) was dissolved in sulfuric acid / water (0.5 mL). Methanesulfonic acid (1.05 g, 10.95 mmol) was added and the mixture was allowed to react at room temperature for 1.5 hours. Methionine (176.6 mg, 1.18 mmol) was then added and the mixture was allowed to react at 40°C overnight. After LCMS monitoring indicated the disappearance of the starting material, the mixture was cooled and the pH was adjusted to neutral with sodium hydroxide and aqueous potassium hydrogen phosphate. The mixture was extracted with ethyl acetate (20 mL x 2). The organic phase was dried and concentrated to dryness to obtain the crude product, which yielded 2-amino-1-(4-fluoro-3-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (5.02 mg). MS (ESI) M / Z: 343.5 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.56 (s, 1H), 7.83 (s, 1H), 7.10 (d, J=11.5Hz, 1H), 6 .83(s, 2H), 6.66(s, 2H), 2.25(d, J=7.7Hz, 6H), 1.76(s, 3H), 1.70(s, 3H).C 18 H 19 FN4O2.

[0496] Example 24

[0497] The compound of Example 23 was prepared by chiral HPLC. SFC separation method: Instrument: SFC-150 mg / m (waters), chiral column: YMC Cellulose-SC (20*250 mm, 5 μm), temperature: 30°C, mobile phase: CO2 / MeOH [0.2% NH3 (7 M in MeOH)] = 65 / 35, flow rate: 50 ml / min, back pressure: 100 bar, detection wavelength: 214 nm, cycle time: 8.68 min. [α] 20 D =-35.95° (c=0.10015, MeOH); >99%ee. MS(ESI)M / Z:343.3[M+H + ]. 1 H NMR (400MHz, DMSO) δ9.53 (s, 1H), 7.82 (s, 1H), 7.09 (d, J=11.5Hz, 1H), 6.8 1(s, 2H), 6.64(s, 2H), 2.26(s, 3H), 2.24(s, 3H), 1.75(s, 3H), 1.70(s, 3H).

[0498] Example 25

[0499]

[0500] The compound of Example 23 was prepared by chiral HPLC. SFC separation method: Instrument: SFC-150 mg / m (waters), chiral column: YMC Cellulose-SC (20*250 mm, 5 μm), temperature: 30°C, mobile phase: CO2 / MeOH [0.2% NH3 (7 M in MeOH)] = 65 / 35, flow rate: 50 ml / min, back pressure: 100 bar, detection wavelength: 214 nm, cycle time: 8.68 min. [α] 20 D =+46.32° (c=0.10125, MeOH); >99%ee. MS(ESI)M / Z:343.3[M+H + ]. 1 H NMR (400MHz, DMSO) δ9.54 (s, 1H), 7.82 (s, 1H), 7.09 (d, J=11.5Hz, 1H), 6.8 1(s, 2H), 6.64(s, 2H), 2.26(s, 3H), 2.24(s, 3H), 1.75(s, 3H), 1.70(s, 3H).

[0501] Example 26

[0502] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(3-hydroxy-3-methylbut-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0503] The synthesis method was the same as that in Example 12, using 6-amino-2-bromo-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide and 2-methylbutyn-3-2-ol as raw materials to prepare 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(3-hydroxy-3-methylbut-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (5.01 mg). MS (ESI) M / Z: 380.1 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.67 (s, 1H), 8.95 (s, 1H), 7.32 (s, 2H), 7.10 (d, J=8.3H z, 1H), 7.01-6.92 (m, 3H), 5.54 (s, 1H), 1.74 (s, 3H), 1.66 (s, 3H), 1.42 (s, 6H).C 20 H 21 N5O3.

[0504] Example 27

[0505] 6-amino-2-(3-fluoro-3-methylpropyl-1-yn-1-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0506] MS (ESI) M / Z: 382.37 [M+H + ].

[0507] Example 28

[0508] 6-amino-4-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0509] The synthesis route of Scheme 1 was adopted. In a single-necked bottle, a chloroform (50 ml) solution of 2-methylpyrimidine-4,6-diol (5 g, 39.65 mmol) was added. The replacement reaction system was a nitrogen system and then cooled to 0 degrees Celsius. After bromine (4.1 ml, 79.3 mmol) was slowly added to the reaction system, the reaction system was stirred at 80 degrees Celsius overnight. After TLC monitoring showed that the raw material disappeared, the reaction solution was concentrated under reduced pressure. The reaction solution was washed with ethyl acetate (100 ml x 3) and filtered to obtain a light yellow solid. The obtained solid was adjusted to pH 7-8 with saturated sodium bicarbonate and dried to give a crude product 5-bromo-2-methylpyrimidine-4,6-diol (11 g). The crude product was then added to phosphorus oxychloride (15 ml) and stirred overnight at 110 degrees Celsius under nitrogen protection. After the reaction, the mixture was diluted with ethyl acetate (100 ml), adjusted to pH 8 with saturated sodium bicarbonate, and then extracted with ethyl acetate (100 ml x 3), washed, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to yield 5-bromo-4,6-dichloro-2-methylpyrimidine (1.2 g). Using 5-bromo-4,6-dichloro-2-methylpyrimidine as starting material, 6-amino-4-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (1.47 mg) was prepared according to the synthetic route of Example 1. MS (ESI) M / Z: 346.1 [M+H] + . 1 H NMR (400MHz, MeOD) δ6.85 (d, J=8.3Hz, 1H), 6.66 (d, J=8.2Hz, 1H), 2.22 (s, 3H), 2.00 (s, 3H), 1.96 (s, 3H).

[0510] Example 29

[0511] 2-Amino-7-fluoro-1-(3-hydroxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[3,2-c]nicotinamide

[0512] Diisopropylamine (319 mg, 3.16 mmol) was dissolved in tetrahydrofuran (10 ml), cooled to -78 degrees Celsius, and 1.8 mol / L n-butyllithium (1.75 ml) was slowly added. The mixture was reacted at -78 degrees Celsius for 15 minutes, followed by a solution of 5-bromo-3-fluoro-2-methylpyridine (500 mg, 2.63 mmol) in tetrahydrofuran (20 ml). The mixture was stirred at this temperature for another 30 minutes, and then a solution of iodine (1 g, 3.95 mmol) in tetrahydrofuran (5 ml) was slowly added, and the mixture was stirred at room temperature for 2 hours. After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to aqueous ammonium chloride solution (80 ml), extracted twice with ethyl acetate (40 ml), and the organic phases were combined. The organic phases were washed twice with sodium thiosulfate (50 ml) and then with saturated brine (40 ml), and then dried over anhydrous sodium sulfate. Finally, the residue was concentrated under reduced pressure and purified by silica gel column chromatography to give 5-bromo-3-fluoro-4-iodo-2-methylpyridine (500 mg).

[0513] 5-Bromo-3-fluoro-4-iodo-2-methylpyridine (470 mg, 1.49 mmol) was dissolved in ethylene glycol dimethyl ether (10 ml), and 3-methoxy-2,6-dimethylaniline (247 mg, 1.64 mmol), cesium carbonate (1.21 g, 3.72 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (77 mg, 0.13 mmol) and tris(dibenzylideneacetone)dipalladium (68 mg, 0.07 mmol) were added. The atmosphere was replaced with nitrogen three times and the reaction solution was stirred at 90 degrees Celsius overnight. After LCMS monitoring showed the disappearance of the starting material, water (30 ml) was added, extracted with ethyl acetate (20 ml), and the combined organic phases were washed with saturated brine (20 ml) and dried over anhydrous sodium sulfate. After concentration under reduced pressure, the product was purified by silica gel column chromatography to give 5-bromo-3-fluoro-N-(3-methoxy-2,6-dimethylphenyl)-2-methylpyridin-4-amine (400 mg).

[0514] 2-Amino-7-fluoro-1-(3-hydroxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[3,2-c]nicotinamide (15.18 mg) was prepared by referring to the protocol of Example 1 using 5-bromo-3-fluoro-N-(3-methoxy-2,6-dimethylphenyl)-2-methylpyridin-4-amine as the starting material. MS (ESI) M / Z: 328.5 [M+H + ]. 1H NMR (400MHz, DMSO-d6) δ9.61 (s, 1H), 8.64 (d, J = 1.6Hz, 1H), 7.06 (d, J = 8.3Hz, 1H) , 6.89 (dd, J=22.5, 6.9Hz, 5H), 2.33 (d, J=3.2Hz, 3H), 1.80 (s, 3H), 1.72 (s, 3H).C 17 H 17 FN4O2.

[0515] Example 30

[0516] 2-Amino-1-(3-hydroxy-2,6-dimethylphenyl)-1H-pyrrolo[3,2-c]pyridine-3-carboxamide

[0517] 2-Amino-1-(3-hydroxy-2,6-dimethylphenyl)-1H-pyrrolo[3,2-c]pyridine-3-carboxamide (5.6 mg) was prepared using 3,4-dibromopyridine as the starting material according to Scheme 5. MS (ESI) M / Z: 297.4 [M+H + ]. 1 H NMR (400MHz, DMSO) δ9.64 (s, 1H), 8.92 (s, 1H), 8.01 (d, J = 5.3Hz, 1H), 7.12 (d, J = 8.3Hz, 1H), 6. 96 (d, J=8.3Hz, 1H), 6.90 (s, 2H), 6.80 (s, 2H), 6.47 (d, J=5.3Hz, 1H), 1.75 (s, 3H), 1.66 (s, 3H).

[0518] Example 31

[0519] 2-amino-1-(3-hydroxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[3,2-c]pyridine-3-carboxamide

[0520] 2-Amino-1-(3-hydroxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[3,2-c]pyridine-3-carboxamide (3.22 mg) was prepared from 5-bromo-4-chloro-2-methylpyridine according to Scheme 5. MS (ESI) M / Z: 311.5 [M+H + ]. 1H NMR (400MHz, DMSO-d6) δ9.65 (s, 1H), 8.77 (s, 1H), 7.11 (d, J=8.3Hz, 1H), 6.95 (d, J=8.3 Hz, 1H), 6.83 (s, 2H), 6.75 (s, 2H), 6.31 (s, 1H), 2.37 (s, 3H), 1.75 (s, 3H), 1.66 (s, 3H).C 17 H 18 N4O2.

[0521] Example 32

[0522] 6-amino-7-(2,6-diethyl-3-hydroxyphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0523] 5-Bromo-4-chloro-2-methylpyrimidine (1 g, 4.83 mmol) and 2,6-diethyl-3-methoxyaniline (690 mg, 3.86 mmol) were dissolved in tetrahydrofuran (30 ml). Lithium bis(trimethylsilyl)amide (1 M, 14.5 ml, 14.5 mmol) was added in an ice-water bath and allowed to react overnight at 50°C. After TLC monitoring indicated the disappearance of the starting material, the mixture was quenched with aqueous ammonium chloride (20 ml) and extracted with ethyl acetate (40 ml). The organic phase was washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The resulting residue was subjected to column chromatography to afford 5-bromo-amine-(2,6-diethyl-3-methoxyphenyl)-2-methylpyrimidin-4-amine (150 mg). 6-Amino-7-(2,6-diethyl-3-hydroxyphenyl)-2-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (15.87 mg) was prepared by using 5-bromo-4-chloro-2-methylpyrimidine and 2,6-diethyl-3-methoxyaniline as raw materials according to the method of Example 1. MS (ESI) M / Z: 339.1 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.60 (s, 1H), 8.87 (s, 1H), 7.12 (d, J=8.4Hz, 1H), 7.06-6.96 (m, 3H), 6.86 (s, 2H), 2.43 (s, 3H), 2.21-1.89 (m, 4H), 0.86 (dt, J=27.7, 7.5Hz, 6H).C 18 H 21 N5O2.

[0524] Example 33

[0525] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0526] 1 H NMR (400MHz, DMSO-d6) δ9.57 (s, 1H), 9.15 (s, 1H), 8.73 (s, 1H), 7.71 (d, J = 8.1Hz, 2H) ,7.18-7.05(m,3H),6.96-6.88(m,3H),6.82-6.69(m,3H),1.83(s,3H),1.74(s,3H).C 21 H 20 N6O2.

[0527] MS (ESI) M / Z: 388.3 [M+H + ].

[0528] Example 34

[0529] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-morpholino-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0530] The preparation method was the same as in Example 13. 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (23.97 mg) was prepared by reacting 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide with morpholine. MS (ESI) M / Z: 383.1 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.54 (s, 1H), 8.66 (s, 1H), 7.06 (d, J = 8.3Hz, 1H), 6.90 (d, J = 8.2Hz, 1H), 6 .79 (s, 2H), 6.70 (s, 2H), 3.59 (t, J=4.8Hz, 4H), 3.42 (t, J=4.8Hz, 4H), 1.79 (s, 3H), 1.70 (s, 3H).C 19 H 22 N6O3.

[0531] Example 35

[0532] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(pyrrolidin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0533] The preparation method was the same as in Example 13. 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (6.97 mg) was prepared by reacting 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide with pyrrolidine. MS (ESI) M / Z: 367.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.52 (s, 1H), 8.59 (s, 1H), 7.05 (d, J=8.3Hz, 1H), 6.89 (d, J= 8.2Hz, 1H), 6.71-6.61(m, 4H), 3.34-3.23(m, 4H), 1.91-1.75(m, 7H), 1.72(s, 3H).C 19 H 22 N6O2.

[0534] Example 36

[0535] 6-amino-2-(dimethylamino)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0536] The preparation method was the same as in Example 13. 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (10.82 mg) was prepared by reacting 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide with dimethylamine hydrochloride. MS (ESI) M / Z: 341.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.53 (s, 1H), 8.60 (s, 1H), 7.06 (d, J=8.2Hz, 1H), 6.89 (d, J=8.2Hz, 1H), 6.71 (s, 2H), 6.65 (s, 2H), 2.93 (s, 6H), 1.80 (s, 3H), 1.71 (s, 3H).C 17 H 20 N6O2.

[0537] Example 37

[0538] 6-amino-2-(4,4-difluoropiperidin-1-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0539] The preparation method was the same as in Example 13. 6-amino-2-(4,4-difluoropiperidin-1-yl)-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (21.39 mg) was prepared by reacting 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide with 4,4-difluoropyridine. MS (ESI) M / Z: 417.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.55 (s, 1H), 8.67 (s, 1H), 7.07 (d, J = 8.3Hz, 1H), 6.90 (d, J = 8.2Hz, 1H), 6.81 (s, 2H), 6.71 (s, 2H), 3.66 (t, J=5.7Hz, 4H), 1.97-1.82 (m, 4H), 1.80 (s, 3H), 1.71 (s, 3H).C 20 H 22 F2N6O2.

[0540] Example 40

[0541] 2,6-Diamino-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0542] 6-Amino-2-chloro-7-(3-methoxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (150 mg, 0.46 mmol) was dissolved in n-butanol (3 ml), and N,N-diisopropylethylamine (177.9 mg, 1.38 mmol) and p-methoxybenzylamine (1.5 ml) were added. The reaction solution was stirred at 150°C for 12 hours. Water (30 ml) was added, and the mixture was extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1) afforded 6-amino-7-(3-methoxy-2,6-dimethylphenyl)-2-((4-methoxybenzyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (168 mg). The product was then dissolved in sulfuric acid / water (13 / 1, 1 ml), and methanesulfonic acid (1.4 g, 14.52 mmol) was added. The reaction was allowed to react at room temperature for 1.5 hours. Methionine (234.3 mg, 1.57 mmol) was then added, and the reaction was allowed to proceed at 40°C overnight. After LCMS monitoring showed the disappearance of the starting material, sodium hydroxide and dipotassium hydrogen phosphate aqueous solution were added to adjust the pH to neutral, and the crude product was directly concentrated to dryness to obtain 2,6-diamino-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (2.01 mg). MS (ESI) M / Z: 313.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.55 (s, 1H), 8.52 (s, 1H), 7.08-7.00 (m, 1H), 6.93 -6.85(m,1H),6.77-6.60(m,4H),5.99(s,2H),1.80(s,3H),1.71(s,3H).C 15 H 16 N6O2.

[0543] Example 41

[0544] 6-amino-2-cyclopropyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0545] 5-Bromo-4-chloro-2-(methylthio)pyrimidine (4 g, 16.81 mmol) was dissolved in tetrahydrofuran (30 ml), and potassium persulfate (15.5 g, 50.42 mmol) was dissolved in water (20 ml) and added dropwise to the reaction mixture. The mixture was stirred at room temperature overnight. After the reaction, the mixture was extracted twice with ethyl acetate (60 ml). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 5-bromo-4-chloro-2-(methylsulfonyl)pyrimidine (4.1 g).

[0546] 5-Bromo-4-chloro-2-(methylsulfonyl)pyrimidine (2 g, 7.33 mmol) was dissolved in tetrahydrofuran (60 ml), and cyclopropylmagnesium bromide (17.6 ml, 8.8 mmol) was added. The reaction mixture was stirred at room temperature overnight. After the reaction, water (30 ml) was added and the mixture was extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 5-bromo-4-chloro-2-cyclopropylpyrimidine (510 mg). Using 5-bromo-4-chloro-2-cyclopropylpyrimidine as the starting material, 6-amino-2-cyclopropyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (5.34 mg) was prepared according to the method of Example 1. MS (ESI) M / Z: 338.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.61 (s, 1H), 8.82 (s, 1H), 7.09 (d, J = 8.3Hz, 1H), 7.02 (s, 2H), 6.94 (d, J = 8.3Hz , 1H), 6.83 (s, 2H), 1.97 (tt, J=7.8, 5.0Hz, 1H), 1.76 (s, 3H), 1.68 (s, 3H), 0.84 (tt, J=7.7, 2.8Hz, 4H).C 18 H 19 N5O2.

[0547] Example 42

[0548] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(2-methylprop-1-en-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0549] The preparation method was the same as in Example 6. 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (70 mg) was prepared by reacting 6-amino-2-chloro-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide with 2-methylprop-1-en-1-yl)boronic acid. MS (ESI) M / Z: 352.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.58 (s, 1H), 8.95 (s, 1H), 7.15 (s, 2H), 7.08 (d, J=8.3Hz, 1H), 6.96-6.85(m, 3H), 6.16(s, 1H), 2.07(s, 3H), 1.84(s, 3H), 1.77(s, 3H), 1.68(s, 3H).C 19 H 21 N5O2.

[0550] Example 43

[0551] 2-amino-1-(4-chloro-3-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0552] 2-Amino-1-(3-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (200 mg, 0.62 mmol) was dissolved in trifluoroacetic acid (6 ml), and N-chlorosuccinimide (197 mg, 1.47 mmol) was added. The mixture was stirred at room temperature. After the reaction, sodium bicarbonate (30 ml) and dichloromethane (30 ml) were added. The mixture was stirred and separated. The organic phase was washed with saturated brine, dried over sodium sulfate, filtered, and the filtrate was concentrated. The crude product was used to prepare 2-amino-1-(4-chloro-3-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (2.44 mg). MS (ESI) M / Z: 359.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ7.81 (s, 1H), 7.15 (s, 1H), 6.74 (s, 2H), 6.63 (s, 2H), 2.25 (d, J = 6.6Hz, 6H), 1.69 (d, J = 10.9Hz, 6H).C 18 H 19 ClN4O2.

[0553] Example 44

[0554] 2-Amino-1-(4-chloro-3-fluoro-5-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0555] Using Scheme 5, the preparation method was the same as in Example 20, using 2,3-dibromo-5,6-dimethylpyridine and 3-fluoro-4-chloro-5-methoxy-2,6-dimethylaniline as starting materials to prepare 2-amino-1-(4-chloro-3-fluoro-5-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (4 mg). MS (ESI) M / Z: 377.1, 379.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ9.82 (s, 1H), 7.85 (s, 1H), 6.95 (s, 2H), 6.67 (s, 2H), 2.26 (d, J=7.0Hz, 6H), 1.74-1.66 (m, 6H).

[0556] Example 45

[0557] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0558] The preparation method was the same as in Example 1, using 5-bromo-4-chloropyrimidine as the starting material to prepare 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (59.74 mg). MS (ESI) M / Z: 298.0 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.62 (s, 1H), 9.00 (s, 1H), 8.41 (s, 1H), 7.21 (s, 2H), 7.09 (d, J = 8.3Hz, 1H), 6.98-6.90 (m, 3H), 1.75 (s, 3H), 1.67 (s, 3H).C 15 H 15 N5O2.

[0559] Example 46

[0560] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-methoxy-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0561] 1-Methoxy-2,4-dimethyl-3-nitrobenzene (3.0 g, 17.95 mmol) was dissolved in dichloromethane (30 ml). A 1 M solution of boron tribromide in dichloromethane (25 ml) was added dropwise at -40°C. The temperature was slowly warmed to room temperature and stirred overnight. After the reaction, water (150 ml) and potassium dihydrogen phosphate (11.2 g) were added to the reaction mixture, which was then extracted with dichloromethane (100 ml). The organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2,4-dimethyl-3-nitrophenol (1.9 g). 2,4-Dimethyl-3-nitrophenol (200.0 mg, 1.3 mmol) and 4-methoxybenzyl alcohol (270.8 mg, 1.9 mmol) were dissolved in tetrahydrofuran, and triphenylphosphine (514.1 mg, 1.9 mmol) was added. Diisopropyl azodicarboxylate was then added dropwise at 0°C, and the mixture was stirred overnight at room temperature. After completion of the reaction, water (20 ml) and ethyl acetate (20 ml) were added to the reaction solution for extraction. The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1-((4-methoxybenzyl)oxy)-2,4-dimethyl-3-nitrobenzene (110 mg). The reaction was then scaled up to prepare 1-((4-methoxybenzyl)oxy)-2,4-dimethyl-3-nitrobenzene (1.06 g).

[0562] Reduced iron powder (1.34 g, 24.0 mmol), silica (2.7 g, 45.6 mmol), and ammonium chloride (790 mg, 14.7 mmol) were added to ethanol (20 ml) and water (20 ml). The reaction system was heated to 60°C. 1-((4-methoxybenzyl)oxy)-2,4-dimethyl-3-nitrobenzene (1.06 g, 3.7 mmol) dissolved in tetrahydrofuran (20 ml) was added to the reaction solution. After stirring at 70°C for 3 hours, saturated brine (30 ml) and ethyl acetate (50 ml) were added to the reaction solution for separation. The organic phase was washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 3-((4-methoxybenzyl)oxy)-2,6-dimethyl-aniline (830 mg). Referring to Example 1, 5-bromo-2,4-dichloropyrimidine (800 mg, 3.5 mmol) was dissolved in N-methylpyrrolidone (10 ml), and 2,6-lutidine (608.5 mg, 5.7 mmol) and 3-((4-methoxybenzyl)oxy)-2,6-dimethyl-aniline (992.5 mg, 3.9 mmol) were added to react and 5-bromo-2-chloro-N-(3-((4-methoxybenzyl)oxy)-2,6-dimethylphenyl)pyrimidin-4-amine (1.0 g) was prepared.

[0563] 5-Bromo-2-chloro-N-(3-((4-methoxybenzyl)oxy)-2,6-dimethylphenyl)pyrimidin-4-amine (400 mg, 0.9 mmol) was dissolved in methanol (8 ml), and sodium methoxide (241.6 mg, 4.5 mmol) was added. The reaction system was heated to 80°C and stirred overnight. After LCMS monitoring showed the disappearance of the starting material, water (30 ml) and ethyl acetate (30 ml) were added to the reaction solution for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 5-bromo-2-methoxy-N-(3-((4-methoxybenzyl)oxy)-2,6-dimethylphenyl)pyrimidin-4-amine (311.1 mg). MS (ESI) M / Z: 446.2 [M+H + ].

[0564] Reference Example 1 6-amino-2-methoxy-7-(3-((4-methoxybenzyl)oxy)-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (60 mg) was prepared by reacting 5-bromo-2-methoxy-N-(3-((4-methoxybenzyl)oxy)-2,6-dimethylphenyl)pyrimidin-4-amine with malononitrile.

[0565] 6-Amino-2-methoxy-7-(3-((4-methoxybenzyl)oxy)-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (60 mg, 0.14 mmol) was dissolved in concentrated sulfuric acid (1 ml). After stirring at room temperature for 1 hour, the reaction mixture was extracted with saturated sodium bicarbonate aqueous solution (20 ml) and ethyl acetate (20 ml). The organic phase was washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 6-Amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-methoxy-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (4.11 mg) was obtained. MS (ESI) M / Z: 328.3 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.61 (s, 1H), 8.73 (s, 1H), 7.08 (d, J=8.3Hz, 1H), 6.96 (dd, J=26.8, 9.2Hz, 3H), 6.82 (s, 2H), 3.77 (s, 3H), 1.77 (s, 3H), 1.68 (s, 3H).C 16 H 17 N5O3.

[0566] Example 47

[0567] 6-amino-2-ethynyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0568] 6-Amino-2-bromo-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (200 mg, 0.535 mmol), triisopropylsilyl acetylene (481 mg, 2.67 mmol), tetrakistriphenylphosphine palladium (124 mg, 0.11 mmol), cuprous iodide (6 mg, 0.005 mmol) were dissolved in N,N-dimethylformamide (6 ml) and triethylamine ( After the reaction, the mixture was extracted with water (20 ml) and ethyl acetate (30 ml), washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting mixture was subjected to column chromatography to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-((triisopropylsilyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (70 mg).

[0569] 6-Amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-((triisopropylsilyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (70 mg, 0.146 mmol) was dissolved in tetrahydrofuran (6 ml), and tetrabutylammonium fluoride (230 mg, 0.732 mmol) was added in an ice-water bath. The mixture was stirred in an ice-water bath for 40 minutes, extracted with water (20 ml) and ethyl acetate (30 ml), washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 6-amino-2-ethynyl-7-(3-hydroxy-2,6-dimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (3.46 mg). MS (ESI) M / Z: 321.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.67 (s, 1H), 8.96 (s, 1H), 7.37 (s, 2H), 7.10 (d, J=8.3Hz, 1H), 7.04-6.93 (m, 3H), 4.03 (s, 1H), 1.75 (s, 3H), 1.66 (s, 3H).C 17 H 15 N5O2.

[0570] Example 48

[0571] 2-Amino-1-(3-chloro-5-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]nicotinamide

[0572] Using Scheme 5, the preparation method was the same as in Example 20, and 2-amino-1-(3-chloro-5-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]nicotinamide (24.61 mg) was prepared from 2,3-dibromo-5,6-dimethylpyridine and 3-chloro-5-methoxy-2,6-dimethylaniline. MS (ESI) M / Z: 359.1 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ10.33 (s, 1H), 7.86 (s, 1H), 7.21 (s, 1H), 6.88 (s, 2H), 6.70 (s, 2H), 2.25 (d, J=7.0Hz, 6H), 1.75 (s, 3H), 1.62 (s, 3H).C 18 H 19 ClN4O2.

[0573] Example 50

[0574] 2-amino-1-(2-hydroxy-3,5-dimethylpyridin-4-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0575] Using the route of Scheme 5 and the same preparation method as in Example 20, 2-amino-1-(2-hydroxy-3,5-dimethylpyridin-4-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (3.17 mg) was prepared from 2,3-dibromo-5,6-dimethylpyridine and 2-methoxy-3,5-dimethylpyridin-4-amine. MS (ESI) M / Z: 326.4 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ7.84 (s, 1H), 7.27 (s, 1H), 7.06 (s, 2H), 6.69 (s, 2H), 2.27 (d, J=5.1Hz, 6H), 1.61 (s, 3H), 1.55 (s, 3H).C 17 H 19 N5O2.

[0576] Example 51

[0577] 2-amino-1-(5-hydroxy-2,4-dimethylpyrimidin-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyrimidine-3-carboxamide

[0578] 1H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 8.15 (s, 1H), 7.85 (s, 1H), 6.97 (s, 2H), 6.69 (s, 2H), 2.25 (d, J=8.5Hz, 6H), 1.94 (s, 3H), 1.72 (s, 3H).C 17 H 19 N5O2.

[0579] MS (ESI) M / Z: 326.2 [M+H + ].

[0580] Example 53

[0581] 2-amino-1-(3-hydroxy-2,5,6-trimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0582] Using the route of Scheme 5, the preparation method was the same as in Example 20. 2-Amino-1-(3-hydroxy-2,5,6-trimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (5.13 mg) was prepared from 2,3-dibromo-5,6-dimethylpyridine and 3-methoxy-2,5,6-trimethylaniline. MS (ESI) M / Z: 339.4 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.40 (s, 1H), 7.83 (d, J = 4.7Hz, 1H), 6.82 (d, J = 5.5Hz, 1H), 6.75-6.56 (m, 4H), 2.30-2.18 (m, 9H), 1.66-1.57 (m, 6H).C 19 H 22 N4O2.

[0583] Example 54

[0584] 2-amino-1-(3,4-difluoro-5-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyrimidine-3-carboxamide

[0585] 1 H NMR (400MHz, DMSO-d6) δ10.16 (s, 1H), 7.85 (s, 1H), 6.93 (s, 2H), 6.69 (s, 2H), 2.26 (d, J=5.7Hz, 6H), 1.71 (s, 3H), 1.67 (s, 3H).C 18 H 18 F2N4O2.

[0586] MS (ESI) M / Z: 359.9 [M+H + ].

[0587] Example 57

[0588] 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-isobutyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide

[0589] 6-Amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-(2-methylprop-1-en-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (60 mg, 0.17 mmol) was dissolved in methanol, 10% Pd / C (3 mg) was added, hydrogen was introduced at normal pressure, and the reaction was carried out at room temperature for 2 hours. After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered and the filtrate was concentrated to give 6-amino-7-(3-hydroxy-2,6-dimethylphenyl)-2-isobutyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (37.03 mg). MS (ESI) M / Z: 354.0 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ9.66 (s, 1H), 8.90 (s, 1H), 7.12-7.03 (m, 3H), 6.94 (d, J=8.2Hz, 1H), 6.85 (s, 2H) , 2.55 (d, J=7.2Hz, 2H), 2.02 (dt, J=13.5, 6.8Hz, 1H), 1.75 (s, 3H), 1.66 (s, 3H), 0.80 (d, J=6.4Hz, 6H).C 19 H 23 N5O2.

[0590] Example 58

[0591] 2-Amino-5-bromo-1-(4-fluoro-3-hydroxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0592] 3,5-Dibromo-2-fluoro-6-methylpyridine (1 g, 3.72 mmol) was dissolved in tetrahydrofuran (10 ml), and 4-fluoro-3-methoxy-2,6-dimethylaniline (628 mg, 3.72 mmol) was added. Lithium hexamethyldisilazide (7.4 ml, 7.44 mmol) was then added dropwise, and the mixture was stirred at room temperature for 0.5 hour. After LCMS monitoring indicated the disappearance of the starting material, ammonium chloride solution (20 ml) was added and stirred for 10 minutes. Ethyl acetate (40 ml) was then added and extracted twice. The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3,5-dibromo-N-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methylpyridin-2-amine (1.2 g). MS (ESI) M / Z: 418.6 [M+H + ].

[0593] Malononitrile (189.5 mg, 2.89 mmol) was dissolved in dioxane (10 ml), sodium tert-butoxide (331 mg, 3.44 mmol) was added, the atmosphere was replaced with nitrogen, and the reaction was carried out at room temperature for 30 minutes. 3,5-dibromo-N-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methylpyridin-2-amine (1.2 g, 2.87 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (234 mg, 0.29 mmol) were added, the atmosphere was replaced with nitrogen, and the reaction was carried out at 120 degrees Celsius for 3 hours. After LCMS monitoring showed the disappearance of the starting material, water (10 ml) was added and extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2-amino-5-bromo-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (499 mg). MS (ESI) M / Z: 402.85 [M+H + ].

[0594] 2-Amino-5-bromo-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (400 mg, 1 mmol) was dissolved in sulfuric acid / water (3 mL). Methanesulfonic acid (3.5 g, 36.8 mmol) was added and the mixture was allowed to react at room temperature for 1.5 hours. Methionine (594 mg, 3.98 mmol) was then added and the mixture was allowed to react at 40°C overnight. After LCMS monitoring indicated the disappearance of the starting material, the mixture was cooled and the pH was adjusted to neutral with sodium hydroxide and aqueous potassium hydrogen phosphate. The mixture was extracted with ethyl acetate (20 mL x 2). The organic phase was dried and concentrated to dryness to obtain the crude product, thereby preparing 2-amino-5-bromo-1-(4-fluoro-3-hydroxy-2,6-dimethylphenyl)-6-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (98.91 mg). 1 H NMR (400MHz, DMSO-d6) δ9.63 (s, 1H), 8.25 (s, 1H), 7.16-7.06 (m, 3H), 6.82 (s, 2H), 2.39 (s, 3H), 1.76 (s, 3H), 1.71 (s, 3H).C 17 H 16 BrFN4O2.MS(ESI)M / Z:407.9[M+H + ].

[0595] Examples 58.1 and 58.2

[0596] SFC method: Instrument: SFC-150mg / m (waters), chiral column: Daicel OZ-3 (25*250mm, 10μm), temperature: 30°C, mobile phase: CO2 / MeOH [0.2% NH3 (7M MeOH)] = 45 / 55, flow rate: 100 mL / min, back pressure: 100 bar, detection wavelength: 214 nm, cycle time: 4.84 min.

[0597] Example 58.1: 40.4 mg. SFC Rt = 1.448 min. [α] D 20 =+35.7°(c=0.10135, MeOH). 1 H NMR (400MHz, DMSO) δ9.59 (s, 1H), 8.25 (s, 1H), 7.11 (d, J = 11.5Hz, 1H), 7.06 (s, 2 H), 6.79(s, 2H), 2.40(s, 3H), 1.76(s, 3H), 1.71(s, 3H).MS(ESI)M / Z: 407.8[M+H + ].

[0598] Example 58.2: 39.8 mg. SFC Rt = 2.031 min. α] D 20 = -16.5° (c = 0.1033 g, MeOH). 1 H NMR (400MHz, DMSO) δ9.60 (s, 1H), 8.25 (s, 1H), 7.11 (d, J = 11.6Hz, 1H), 7.06 (s, 2 H), 6.79(s, 2H), 2.39(s, 3H), 1.76(s, 3H), 1.71(s, 3H).MS(ESI)M / Z: 407.8[M+H + ].

[0599] Example 59

[0600] 2-amino-1-(4-fluoro-3-hydroxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0601] In a one-necked flask, 6-chloro-5-(trifluoromethyl)pyridin-2-amine (39 g, 0.19 mol) was dissolved in dioxane / water (10:1). Methylboric acid (23.4 g, 0.39 mol), potassium carbonate (82 g, 0.59 mol), and PdCl2(dppf) (8 g, 0.011 mol) were then added. The reaction system was replaced with nitrogen and stirred at 110°C for 16 hours. LCMS analysis indicated the disappearance of the starting material and the formation of the product. The reaction mixture was quenched with purified water and extracted with ethyl acetate (700 mL x 3). The organic phases were combined, washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to yield 6-methyl-5-(trifluoromethyl)pyridin-2-amine (24 g).

[0602] In a single-necked flask, 6-methyl-5-(trifluoromethyl)pyridin-2-amine (24 g, 0.14 mol) was dissolved in acetonitrile (300 ml). N-bromosuccinimide (29.13 g, 0.164 mol) was added portionwise under ice-cooling. The reaction system was stirred at room temperature for 16 hours. LCMS analysis showed the disappearance of the starting material and the formation of the product. The reaction solution was poured into 500 ml of water. Extraction was performed with ethyl acetate (400 ml x 3). The organic phases were combined and washed with saturated brine (100 ml x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to give 3-bromo-6-methyl-5-(trifluoromethyl)pyridin-2-amine (26.3 g). MS (ESI) M / Z: 257.0 [M+H + ]. 1H NMR (400MHz, CDCl3) δ8.04 (s, 1H), 5.30 (s, 1H, -NH2), 2.73-2.59 (m, 3H).

[0603] In a three-necked flask, 3-bromo-6-methyl-5-(trifluoromethyl)pyridin-2-amine (26.3 g, 0.103 mol) was dissolved in bromoform, and bromine (19.9 g, 0.124 mol) was slowly added dropwise at 0 degrees Celsius. Tert-butyl nitrite (32.61 g, 0.31 mol) was then slowly added dropwise to the reaction solution at 0 degrees Celsius. The mixture was reacted at room temperature for 3 hours. LCMS detected that the raw material disappeared and the product was generated. The reaction solution was quenched by adding ice water, and the organic phase was washed three times with saturated sodium bicarbonate solution. The obtained organic phase was concentrated under reduced pressure using an oil pump to remove bromoform. The obtained mixture was purified by silica gel column chromatography to give 2,3-dibromo-6-methyl-5-(trifluoromethyl)pyridine (26.5 g). MS (ESI) M / Z: 320.0 [M+H + ].

[0604] In a single-necked flask, 2,3-dibromo-6-methyl-5-(trifluoromethyl)pyridine (15.1 g, 0.047 mol) was dissolved in ethylene glycol dimethyl ether (150 mL). 4-Fluoro-3-methoxy-2,6-dimethylaniline (8.8 g, 0.053 mol), cesium carbonate (38.75 g, 0.12 mol), Pd2dba3 (4.34 g, 4.74 mmol), and XantPhos (5.48 g, 9.48 mmol) were added at room temperature. The reaction system was replaced with nitrogen. The reaction system was reacted at 110°C for 12 hours. LCMS analysis indicated the disappearance of the starting material and the formation of the product. The reaction solution was quenched with purified water and the mixture was extracted with ethyl acetate (400 mL x 3). The organic phases were combined, washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to give 3-bromo-N-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)pyridin-2-amine (10.5 g). MS (ESI) M / Z: 407.1 [M+H] + .

[0605] To a 250 ml three-necked flask, sodium tert-butoxide (12.42 g, 129.3 mmol) was added. Ethylene glycol dimethyl ether (30 ml) was then added under nitrogen. Then, a solution of malononitrile (8.53 g, 129.3 mmol) in ethylene glycol dimethyl ether (5 ml) was added dropwise at room temperature. After the addition was complete, the reaction mixture was stirred at room temperature for 30 minutes. Then, under nitrogen, a solution of 3-bromo-N-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)pyridin-2-amine (10.5 g, 25.86 mmol) in ethylene glycol dimethyl ether (30 ml) and PdCl2(dppf) (1.89 g, 2.586 mmol) were added. The reaction system was allowed to react at 110 degrees Celsius for 16 hours. LCMS analysis indicated the formation of product, and the reaction mixture was poured into ice water. The reaction mixture was extracted with ethyl acetate (300 ml x 3), and the organic phases were combined, washed with saturated brine (50 ml x 3 times), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to give 2-amino-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (5.7 g, yield 56.3%). MS (ESI) M / Z: 393.2 [M+H] + .

[0606] 2-Amino-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (5.7 g, 14.55 mmol) was dissolved in concentrated sulfuric acid (50 ml) under ice. The reaction mixture was stirred at room temperature for 2 hours. After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was poured into ice water and the pH was adjusted to 8 with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate (30 ml x 3). The organic phases were combined, washed with saturated brine (3 times with 50 ml), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting mixture was purified by silica gel column chromatography to yield 2-amino-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (4.4 g). 1 H NMR (400MHz, DMSO) δ8.28 (s, 1H), 7.26 (d, J = 12.2Hz, 1H), 7.19 (s, 2H), 6.94 (s, 2H), 3.87 ( d, J=1.0Hz, 3H), 2.45 (d, J=1.6Hz, 3H), 1.82 (s, 3H), 1.78 (s, 3H). MS (ESI) M / Z: 411.3 [M+H] + .

[0607] To a 250 ml three-necked flask, add 2-amino-1-(4-fluoro-3-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (4.4 g, 10.7 mmol) and anhydrous dichloromethane (40 ml). Add a 1N dichloromethane solution of BBr3 dropwise under ice-cooling, and stir the reaction mixture at room temperature for 2 hours. LCMS analysis indicated the disappearance of the starting material and the formation of the product. The reaction mixture was quenched with ice water and the pH was adjusted to 9 with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate (20 ml x 3). The organic phases were combined, washed with saturated brine (50 ml, 3 times), dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The resulting residue was purified by acetonitrile slurrying to give 2-amino-1-(4-fluoro-3-hydroxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (3.1 g). 1 H NMR (400MHz, DMSO) δ9.63 (s, 1H), 8.27 (s, 1H), 7.19-7.07 (m, 3H), 6.93 (s, 2 H), 2.45 (d, J=1.5Hz, 3H), 1.77 (s, 3H), 1.72 (s, 3H). MS (ESI) M / Z: 397.1[M+H + ].

[0608] Examples 59.1 & 59.2

[0609] SFC method: Instrument: SFC-150mg / m (waters), chiral column: Daicel OZ (25*250mm, 10μm), temperature: 30°C, mobile phase: CO2 / MeOH [0.2% NH3 (7M MeOH)] = 65 / 35, flow rate: 100 ml / min, back pressure: 100 bar, detection wavelength: 214 nm, cycle time: 10 min.

[0610] Example 59.1: 2.13 mg. SFC Rt = 1.770 min. [α] D 20 =+42.4°(0.10018, MeOH). 1 H NMR (400MHz, DMSO) δ9.63 (s, 1H), 8.26 (s, 1H), 7.19-7.05 (m, 3H), 6.92 (s, 2 H), 2.45 (d, J=1.6Hz, 3H), 1.77 (s, 3H), 1.72 (s, 3H). MS (ESI) M / Z: 397.1[M+H + ].

[0611] Example 59.2: 2.36 mg. SFC Rt = 2.480 min. [α] D 20 =-26.0°(0.10100, MeOH). 1 H NMR (400MHz, DMSO) δ9.63 (s, 1H), 8.26 (s, 1H), 7.13 (t, J=5.6Hz, 3H), 6.92 (s, 2H), 2.45(d, J=1.6Hz, 3H), 1.76(s, 3H), 1.72(s, 3H).MS(ESI)M / Z: 397.1[M+H + ].

[0612] Example 120

[0613] Reaction route:

[0614] 6-Chloro-5-(trifluoromethyl)pyridin-2-amine (1 g, 5.01 mmol) was dissolved in dioxane / water (10 / 1, 11 ml). Methylboric acid (910.6 mg, 15.2 mmol), potassium carbonate (2.1 g, 15.2 mmol), and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (206.6 mg, 0.25 mmol) were added. The atmosphere was purged with nitrogen three times, and the reaction solution was stirred at 100°C overnight. After LCMS monitoring indicated the disappearance of the starting material, water (30 ml) was added and the mixture was extracted twice with ethyl acetate (30 ml). The organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 6-methyl-5-(trifluoromethyl)pyridin-2-amine (710 mg). MS (ESI) M / Z: 177.0 [M+H + ].

[0615] 6-Methyl-5-(trifluoromethyl)pyridin-2-amine (710 mg, 4.03 mmol) was dissolved in acetonitrile (8 ml), and N-bromosuccinimide (861.7 mg, 4.84 mmol) was added. The mixture was stirred at room temperature overnight. After LCMS monitoring showed the disappearance of the starting material, an aqueous sodium thiosulfate solution (20 ml) was added. The mixture was extracted twice with ethyl acetate (40 ml). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3-bromo-6-methyl-5-(trifluoromethyl)pyridin-2-amine (880 mg). MS (ESI) M / Z: 254.7 [M+H + ].

[0616] 3-Bromo-6-methyl-5-(trifluoromethyl)pyridin-2-amine (580 mg, 2.28 mmol) was dissolved in bromoform (4 ml), and liquid bromine (301 mg, 1.9 mmol) was added. Tert-butyl nitrite (500 mg, 4.74 mmol) was added dropwise, and the mixture was stirred at room temperature for 2 hours. After LCMS monitoring showed the disappearance of the starting material, water (30 ml) was added and the mixture was extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2,3-dibromo-6-methyl-5-(trifluoromethyl)pyridine (280 mg). 1 H NMR (400MHz, CDCl3) δ8.04 (s, 1H), 2.66 (s, 3H).C7H5BrF3NO.

[0617] 2,3-Dibromo-6-methyl-5-(trifluoromethyl)pyridine (280 mg, 0.92 mmol), 3-fluoro-5-methoxy-2,6-dimethylaniline (155 mg, 0.92 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (48 mg, 0.083 mmol), cesium carbonate (748 mg, 2.3 mmol) and trisdibenzylideneacetone dipalladium (42 mg, 0.046 mmol) were added to ethylene glycol dimethyl ether (15 ml), the atmosphere was replaced with nitrogen three times, and the reaction was carried out at 90°C overnight. After LCMS monitoring showed the disappearance of the starting material, water (30 ml) was added and extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 3-bromo-N-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)pyridin-2-amine (40 mg). MS (ESI) M / Z: 407.0 [M+H + ].

[0618] Malononitrile (20 mg, 0.3 mmol) was dissolved in dioxane (5 ml), sodium tert-butoxide (50 mg, 0.5 mmol) was added, the atmosphere was replaced with nitrogen, and the reaction was carried out at room temperature for 30 minutes. 3-Bromo-N-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)pyridin-2-amine (40 mg, 0.1 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (8.15 mg, 0.01 mmol) were added, the atmosphere was replaced with nitrogen, and the reaction was carried out at 120 degrees Celsius for 3 hours. After LCMS monitoring showed the disappearance of the starting material, water (10 ml) was added and extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 2-amino-1-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (40 mg). MS (ESI) M / Z: 393.2 [M+H + ].

[0619] 2-Amino-1-(3-fluoro-5-methoxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (40 mg, 0.102 mmol) was dissolved in sulfuric acid / water (0.15 ml), and methanesulfonic acid (400 mg, 3.78 mmol) was added. The mixture was reacted at room temperature for 1.5 hours, and methionine (61 mg, 0.41 mmol) was added. The mixture was reacted at 40°C overnight. After LCMS monitoring showed the disappearance of the starting material, the mixture was cooled, the pH was adjusted to neutral with sodium hydroxide and dipotassium hydrogen phosphate aqueous solution, and extracted with ethyl acetate (20 ml * 2). The organic phase was dried and concentrated to dryness to obtain a crude product. The crude product was used to prepare 2-amino-1-(3-fluoro-5-hydroxy-2,6-dimethylphenyl)-6-methyl-5-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (2.27 mg). MS (ESI) M / Z: 397.0 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ 10.09 (s, 1H), 8.28 (s, 1H), 7.18 (s, 2H), 6.96 (s, 2H), 6.83 (d, J = 11.1Hz, 1H), 2.46 (s, 3H), 1.65 (d, J = 16.0Hz, 6H).

[0620] Example 189

[0621] Reaction route:

[0622] 5-Bromoisoquinolin-8-amine (5.0 g, 22.4 mmol), methylboronic acid (4.0 g, 67.2 mmol), and potassium carbonate (9.27 g, 67.2 mmol) were dissolved in dioxane (100 ml) and water (10 ml), followed by the addition of [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (900 mg, 1.1 mmol). The atmosphere was purged with nitrogen three times, and the reaction mixture was stirred at 100°C overnight. After TLC monitoring indicated the disappearance of the starting material, water (100 ml) was added, and the mixture was extracted twice with ethyl acetate (100 ml). The organic phases were combined, washed with saturated brine (80 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 5-methylisoquinolin-8-amine (3.5 g). MS (ESI) M / Z: 158.7 [M+H + ].

[0623] 5-Methylisoquinolin-8-amine (3.5 g, 27.2 mmol) was dissolved in acetonitrile (80 ml) and N-bromosuccinimide (4.85 g, 27.2 mmol) was added. The reaction was stirred at room temperature overnight. After TLC monitoring showed the disappearance of the starting material, water (100 ml) was added and the mixture was extracted twice with ethyl acetate (100 ml). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 7-bromo-5-methylisoquinolin-8-amine (3.2 g). MS (ESI) M / Z: 237.2 [M+H + ]. 1 H NMR (400MHz, CDCl3) δ9.33 (s, 1H), 8.56 (d, J = 5.9Hz, 1H), 7.67 (d, J = 5.9Hz, 1H), 7.54 (s, 1H), 4.80 (s, 2H), 2.53 (s, 3H).C 10 H9BrN2.

[0624] 7-Bromo-5-methylisoquinolin-8-amine (1.5 g, 6.3 mmol) was dissolved in hydrochloric acid (37%, 7 ml) and water (1.8 ml). The temperature was lowered to -15°C, and a solution of sodium nitrite (480 mg, 6.9 mmol) in water (8.4 ml) was added dropwise. After the addition was complete, the mixture was stirred at 0°C for 30 minutes. The above reaction solution was slowly added dropwise to a solution of potassium iodide (9.4 g, 56.7 mmol) in water (84 ml). After the addition was complete, the mixture was stirred at room temperature overnight. After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and quenched with saturated sodium sulfite solution (30 ml) and saturated sodium bicarbonate solution (30 ml). The mixture was extracted with dichloromethane (50 ml x 2). The organic phase was washed with brine, dried over sodium sulfate, and concentrated to dryness to obtain the crude product. The residue was concentrated under reduced pressure and purified by silica gel column chromatography to yield 7-bromo-8-chloro-5-methylisoquinoline (1.1 g). 1 H NMR (400MHz, CDCl3) δ9.55 (s, 1H), 8.65 (d, J=5.8Hz, 1H), 7.68 (d, J=5.9Hz, 1H), 7.62 (s, 1H), 2.65 (s, 3H).

[0625] 7-Bromo-8-chloro-5-methylisoquinoline (1.1 g, 4.26 mmol), 3-methoxy-2,6-dimethylaniline (600 mg, 4.26 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (240 mg, 0.43 mmol), and cesium carbonate (3.4 g, 10.6 mmol) were dissolved in toluene (22 ml), and tris(dibenzylideneacetone)dipalladium (180 mg, 0.2 mmol) was added. The reaction mixture was purged with nitrogen three times, and the reaction was allowed to proceed at 100°C overnight. After TLC monitoring showed that the starting material disappeared, the mixture was cooled, water (50 ml) was added, and the mixture was extracted with ethyl acetate (20 ml * 2). The organic phase was washed with brine, dried over sodium sulfate, and concentrated to dryness to obtain a crude product. Finally, the residue was concentrated under reduced pressure and purified by silica gel column chromatography to obtain 8-chloro-N-(3-methoxy-2,6-dimethylphenyl)-5-methylisoquinolin-7-amine (1.3 g). MS (ESI) M / Z: 327.39 [M+H + ].

[0626] Malononitrile (728.83 mg, 11.04 mmol) was dissolved in N,N-dimethylformamide (15 ml), sodium tert-butoxide (1.59 g, 16.56 mmol) was added, the atmosphere was replaced with nitrogen, and the reaction was carried out at room temperature for 30 minutes. 8-chloro-N-(3-methoxy-2,6-dimethylphenyl)-5-methylisoquinolin-7-amine (900 mg, 2.76 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (225 mg, 0.28 mmol) were added, the atmosphere was replaced with nitrogen, and the reaction was carried out at 130°C for 3 hours. After LCMS monitoring showed the disappearance of the starting material, water (30 ml) was added and extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give 8-amino-7-(3-methoxy-2,6-dimethylphenyl)-5-methyl-7H-pyrrolo[2,3-h]isoquinoline-9-carbonitrile (420 mg). MS (ESI) M / Z: 357.39 [M+H + ].

[0627] 8-Amino-7-(3-methoxy-2,6-dimethylphenyl)-5-methyl-7H-pyrrolo[2,3-h]isoquinoline-9-carbonitrile (420 mg, 1.18 mmol) was dissolved in sulfuric acid (5 ml) and allowed to react at room temperature for 1 hour. After TLC monitoring showed the disappearance of the starting material, the mixture was cooled and the pH was adjusted to neutral with aqueous sodium hydroxide solution. The mixture was extracted with ethyl acetate (30 ml x 2). The organic phase was dried and concentrated to dryness to obtain a crude product. Finally, the crude product was concentrated under reduced pressure to obtain 8-amino-7-(3-methoxy-2,6-dimethylphenyl)-5-methyl-7H-pyrrolo[2,3-h]isoquinoline-9-carboxamide (500 mg).

[0628] 8-Amino-7-(3-methoxy-2,6-dimethylphenyl)-5-methyl-7H-pyrrolo[2,3-h]isoquinoline-9-carboxamide (500 mg, 1.37 mmol) was dissolved in dichloromethane (20 ml), and a 2M solution of boron tribromide in dichloromethane (4 ml, 8.0 mmol) was added. The mixture was stirred at room temperature overnight. After LCMS monitoring showed the disappearance of the starting material, aqueous sodium bicarbonate (50 ml) was slowly added, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain 8-amino-7-(3-hydroxy-2,6-dimethylphenyl)-5-methyl-7H-pyrrolo[2,3-h]isoquinoline-9-carboxamide (3.36 + 52.84 mg). MS (ESI) M / Z: 361.44 [M+H + ]. 1H NMR (400MHz, DMSO-d6) δ9.79 (s, 1H), 8.26-8.16 (m, 2H), 8.15 (s, 1H), 7.76 (d, J=5.9Hz, 1H), 7.23 (d, J =8.3Hz, 1H), 7.07 (d, J = 8.2Hz, 1H), 6.93 (s, 2H), 6.53 (s, 2H), 2.67 (s, 3H), 1.77 (s, 3H), 1.68 (s, 3H).

[0629] The preparation of the following compounds refers to the preparation method of the above examples. [Corrected 02.02.2024 in accordance with Article 91]

[0630] Biological activity test method:

[0631] Biological Test Example 1. Enzyme Inhibitory Activity of the Disclosed Compounds

[0632] The enzymatic activity of some of the example compounds was verified using the ADP-GLO method

[0633] Experimental Materials:

[0634] PKMYT1 enzyme protein was purchased from Thermo Scientific (Cat. No. A33387), and ADP-GLO kit was purchased from Promega (Cat. No. V9103).

[0635] PKMYT1 enzyme activity reaction buffer: 70 mM HEPES, 3 mM MgCl2, 3 mM MnCl2, 50 μg ml-1 PEG20000, 3 μM sodium orthovanadate, 1.2 mM DTT.

[0636] Experimental steps:

[0637] Using an ultra-micropipette, test compound (3.33 mM, dissolved in DMSO) was diluted sequentially in a 1:3 gradient (10,000 nM, 3,333 nM, 1,111 nM, 370 nM, 123 nM, 41 nM, 13.7 nM, 4.6 nM, 1.5 nM, and 0.5 nM) starting at 10 uM. The dilutions were then added to a 384-well plate, with two replicates for each concentration. Positive and negative controls (16 wells each) were set up, with replicates containing 1 uM of the positive compound for the positive control and DMSO for the negative control. The plate was centrifuged at 2,500 rpm for 1 minute.

[0638] Add 5 μL of the prepared enzyme solution to each well of the reaction plate for a final enzyme concentration of 10 nM. Centrifuge at 1000 rpm for 1 minute. Incubate at 37°C for 15 minutes.

[0639] Add 5 μL of the prepared ATP substrate solution to the reaction plate, for a final ATP concentration of 5 μM. Centrifuge at 1000 rpm for 1 minute. Seal the plate with aluminum foil and place in an incubator at 37°C for 90 minutes.

[0640] Add 10uL ADP-glo TM Use a reagent to stop the kinase reaction and deplete unused ATP, leaving only ADP and a very low background ATP. Centrifuge at 1000 rpm for 1 minute. Seal the plate with aluminum foil and incubate at room temperature for 60 minutes.

[0641] Add 20 μL of kinase assay reagent to convert ADP to ATP. Introduce luciferase and luciferin to detect ATP. Centrifuge at 1000 rpm for 1 minute. Seal the plate with aluminum foil and incubate at room temperature for 60 minutes.

[0642] ADP-GLO luminescence signal values ​​were read and analyzed using a PHERAstar microplate reader. The average inhibition rate of the positive control replicates was set as 100% relative inhibition rate; the average inhibition rate of the negative control replicates was set as 0% relative inhibition rate. The ADP-GLO readings were converted to relative inhibition rates, and the compound IC values ​​were fitted using a four-parameter model. 50 .

[0643] Experimental results:

[0644] This indicates that the compound has good enzyme inhibitory activity. For specific results, see Table 1.

[0645] Table 1 PKMYT1 enzyme activity test results

[0646] “ / ” indicates not tested or poor activity.

[0647] Biological Test Example 2. Cytostatic Activity of the Disclosed Compounds

[0648] The biological activities of some of the compounds of the present invention were verified using CTG assay (HCC1569).

[0649] Experimental Materials:

[0650] HCC1569 cells were purchased from ATCC (Cat. No. CRL-2330) and cultured in a 37°C, 5% CO2 cell culture incubator. HCC1569 complete culture medium: RPMI-1640 liquid medium (Cat. No. Gibico 11875-093), 20% FBS (Cat. No. Gibico 10099-141), and 1% Pen Strep (Cat. No. Gibico 15070-063).

[0651] Test compound: Compound of the Examples of the present disclosure.

[0652] Experimental steps:

[0653] 75cm 2 HCC1569 cells in a culture flask were digested with 2 mL of trypsin for 2-3 minutes, then neutralized with 2 mL of 1640 complete medium. Centrifuge at 1200 rpm for 5 minutes. Resuspend the cells in 4 mL of 1640 complete medium. Count 500 μL of the cell suspension using a Vi-CELL-XR cell counter.

[0654] Using a Multidrop instrument, 1000 HCC1569 cells were seeded per well (50 μL 1640 Growth Media) in a 384-well plate, and drugs were added 24 hours later.

[0655] Using an ultra-micropipette, the test compound (concentration: 3.33 mM, dissolved in DMSO) was diluted in 10 gradients (10000 nM, 3333 nM, 1111 nM, 370 nM, 123 nM, 41 nM, 13.7 nM, 4.6 nM, 1.5 nM, 0.5 nM) with a maximum starting concentration of 10 uM, and the drug was added. Two replicate wells were set for each concentration; 14 wells each for positive and negative controls were set, the positive control was a replicate well of 10 uM positive compound, and the negative control was a replicate well of DMSO.

[0656] After drug addition, the cells were placed in a 37°C incubator and cultured for 5 days. After 5 days, 25ul of CTG buffer was added to each well for CTG assay detection, and the plate was read using a microplate reader for analysis. The CTG readings were analyzed. The average inhibition rate of the positive control replicates was set as the relative inhibition rate of 100%; the average inhibition rate of the negative control replicates was set as the relative inhibition rate of 0%. The CTG readings were converted to relative inhibition rates, and the inhibition rate (inhibition%) of each concentration of the test compound on the cells was converted according to the following formula: Inhibition% = (bx) / (ba) * 100%;

[0657] a = CTG value (highest concentration), b = CTG value (blank well), x = CTG value (test well value).

[0658] IC was analyzed using GraphPad PRISM 8. 50 Perform calculations.

[0659] (1) Statistically analyze the concentrations and inhibition rates corresponding to 10000nM, 3333nM, 1111nM, 370nM, 123nM, 41nM, 13.7nM, 4.6nM, 1.5nM, and 0.5nM. Statistically calculate the data using Log10 (A compound concentration). (2) Input the data into GraphPad PRISM 8 and select Analysis. (3) Select Nonlinear regression (curve fit). (4) Select Log (inhibitor) vs. response-Variable slope. (5) Select the calculation formula and calculate it according to the following formula; Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope)), X: Dose or concentration logarithm, Y: Response value, Top and Bottom: Peak and bottom values. (6) Fit the data to obtain IC 50 (7) Adjust the fitting conditions according to the specific data. Adjust the constraints of Bottom, Top and HillSlope appropriately. The curve that best fits the actual situation has been achieved.

[0660] The experimental results show that the compound has good cell inhibitory activity. The specific results are shown in Table 2

[0661] Table 2 CTG test results

[0662] IC of cells by the compounds of the present disclosure 50 Preferably not more than 1 μM, preferably not more than 0.5 μM, preferably not more than 0.1 μM, preferably not more than 0.05 μM, more preferably not more than 0.02 μM.

[0663] Biological Test Example 3: Study on the Metabolic Stability of Compounds in Mouse Liver Microsomes

[0664] The concentration of the parent drug in the incubation system was determined using LC / MS / MS, and the intrinsic clearance of the test compound in the microsomal system was calculated to evaluate its stability. The test concentration of the test compound and the positive control compound was 1 μM.

[0665] 1. Transfer 25 μL of NADPH (10 mM) or phosphate buffered saline (100 mM, pH 7.4) to the liver microsome incubation system and add 2 μL of the test substance or verapamil at a concentration of 200 μM. Prepare duplicates for samples with NADPH; prepare single replicates for samples without NADPH.

[0666] 2. Take 30 μl of the suspension at 0.5, 5, 15, 30, and 60 minutes, add 180 μl of acetonitrile containing internal standard to terminate the reaction, and vortex for 10 minutes.

[0667] 3. Centrifuge at 3220g for 20 minutes to precipitate the protein. Refrigerate the plate at 4°C for 30 minutes, then re-centrifuge at 3220g for 20 minutes. Transfer 100 μl of the supernatant to the sample plate and add 100 μl of pure water to mix thoroughly before analysis by UPLC-MS / MS.

[0668] 4. All data calculations were performed using Microsoft Excel. Peak areas were detected by extracting ion spectra. The in vitro half-life (t 1 / 2 ).

[0669] In vitro half-life (t 1 / 2 ) calculated by slope: in vitro t 1 / 2 =0.693 / k

[0670] In vitro clearance Cl int (Unit: μl / min / mg) Calculation: in vitro C Lint = kV / N; V = incubation volume per well (400 μl); V = incubation volume per well (400 μl). Specific results are shown in Table 3.

[0671] Positive control: RP-6306, its structure is as follows:

[0672] Table 3 Results of metabolic stability test of mouse liver microsomes

[0673] Conclusion: Compared with RP-6306, the disclosed compound has a significantly longer half-life, a slower clearance rate, and better metabolic stability.

[0674] Biological Test Example 4 Tissue Distribution of the Disclosed Compounds

[0675] OVCAR3 tumor cells (ATCC, catalog number HTB-161) were cultured in RPMI 1640 medium containing 20% ​​inactivated fetal bovine serum, supplemented with 0.01 mg / mL insulin and 1% penicillin-streptomycin. Cultures were maintained in a 37°C, 5% CO2 incubator. The medium was changed every other day, and cells were passaged every 3 to 4 days after confluence. Under sterile conditions, the in vitro cultured OVCAR3 cell suspension was centrifuged and then added to adjust the cell concentration to 1×10 8ceils / mL, an equal volume of Matrigel was added and inoculated subcutaneously on the back of the right hind forelimb of mice (BALB / c nude mice, female, Beijing Weitong Lihua Experimental Animal Co., Ltd.) (0.1 mL / mouse). 31 days after inoculation, the average tumor volume was 100-150 mm 3 Mice were randomly divided into groups according to tumor size and body weight, and the administration of test compounds was started.

[0676] The experiment was divided into a solvent control group, a positive control group, and a test group, with 5-6 mice in each group. The specific experimental protocol and the frequency of dosing for each group are shown in the table below. For the test compound-administered group, the test compound was mixed in 0.5% methylcellulose and administered orally twice daily. The experiment was terminated after 28 days of dosing. PK plasma (EDTA-K2 anticoagulation) samples were collected (0.5h, 1h, 2h, and 6h after dosing). Mice were euthanized and tumor samples were collected 2h and 6h after the last dose.

[0677] Tumor concentration test method:

[0678] All tumor samples were added to pure water at a ratio of 1:3 (3 mL of solvent was added per g of tumor) and ground using a frozen homogenizer to obtain tumor sample solutions.

[0679] Standard curve preparation method:

[0680] The test compound powder was diluted in a gradient with 50% acetonitrile in water to prepare a series of working solutions. 3 μL of the working solution (10, 20, 40, 100, 200, 1000, 2000, 10,000, 20,000 ng / mL) was added to 57 μL of a blank Balb / c nude mouse tumor sample solution to obtain calibration standard solutions ranging from 0.5 to 1000 ng at concentrations of 0.5, 1, 2, 5, 10, 50, 100, 500, and 1000 ng / mL, respectively, in a total volume of 60 μL.

[0681] Quality control samples were prepared in the same manner as the calibration standards: 3 μL of working solution (30, 60, 120, 1000, 8000, 16000 ng / mL) was added to 57 μL of blank Balb / c nude mouse tumor sample solution to obtain 6 quality control samples with concentrations of 1.5 ng / mL, 3 ng / mL, 6 ng / mL, 50 ng / mL, 400 ng / mL and 800 ng / mL, respectively, in a total volume of 60 μL.

[0682] Take 30 μL of sample (including standard solution, quality control sample, and test sample) and add 200 μL of protein precipitant containing acetonitrile to precipitate protein. Vortex for 30 seconds, then centrifuge at 3900 rpm at 4°C for 15 minutes. Aspirate the supernatant and dilute it 3-fold with water. 5 μL of the diluted supernatant is loaded onto the LC / MS / MS system for quantitative analysis.

[0683] Dosage and frequency of each group

[0684] The experimental results are shown in Table 4:

[0685] Table 4 Tumor tissue concentration test results of the disclosed compounds

[0686] The above experimental results show that the tumor tissue concentration of the disclosed compound 2 hours and 6 hours after administration is significantly better than that of the positive drug, indicating that the disclosed compound has potentially better anti-tumor activity.

[0687] Biological Test Example 5 Metabolic Stability of the Disclosed Compounds in Human Hepatocytes

[0688] Prepare several 96-well sample precipitation plates, named T0, T15, T30, T60, T90, T120, T240, T0-MC, T240-MC, and blank matrix. Remove the recovery medium and incubation medium in advance and place them in a 37°C water bath to preheat. Remove the frozen hepatocytes from the liquid nitrogen tank and immediately immerse them in a 37°C water bath (about 90 seconds). After the frozen portion is thawed and loosened, pour them into centrifuge tubes containing 40mL of recovery medium and gently invert to resuspend the cells in the recovery medium. Centrifuge at 100×g for 5 minutes at room temperature, remove the supernatant, resuspend the hepatocytes in an appropriate volume of incubation medium, and calculate the cell viability using trypan blue staining. 198 μL of hepatocyte suspension (0.51×106 cells / mL) was added to the preheated incubation plate. For the culture medium control group, 198 μL of incubation medium without hepatocytes was added to the T0-MC and T120-MC incubation plates. All incubation plates were preincubated in a 37°C incubator for 10 minutes.

[0689] Add 2 μL of the test and control compound working solutions, mix thoroughly, and immediately place the plate on a plate shaker in the incubator. Start the timer to initiate the reaction. Prepare two replicates for each compound at each time point. Incubate at 37°C, saturated humidity, and 5% CO2.

[0690] In the assay system, the final concentration of the test article was 1 μM, the final concentration of the control article was 3 μM, the final concentration of hepatocytes was 0.5 × 10⁶ cells / mL, and the final concentration of total organic solvent was 0.96%, including 0.1% DMSO. At the end of the incubation period, the incubation plates were removed, and 25 μL of the mixture of compound and control compound with cells was added to a sample plate containing 125 μL of stop solution (200 ng / mL tolbutamide and labenolol in acetonitrile). For the blank sample plate, 25 μL of the incubation medium without hepatocytes was added directly. All sample plates were sealed and shaken on a plate shaker at 600 rpm for 10 minutes, followed by centrifugation at 3220 × g for 20 minutes. The supernatants of the test article and control articles were diluted 1:3 with ultrapure water. All samples were mixed and analyzed by LC / MS / MS.

[0691] The experimental results are shown in Table 5:

[0692] Table 5 Human HMS metabolic stability of the disclosed compounds

[0693] The experimental results disclosed herein show significant gender differences in the metabolism of RP-6306 in human hepatocytes in vitro, with male hepatocytes exhibiting significantly lower metabolic stability than female hepatocytes. However, the disclosed compounds exhibit no gender differences in metabolic stability in human hepatocytes in vitro, exhibiting good stability. This reduces the risk of gender-related effects on drug efficacy and safety, facilitating more convenient clinical applications.

[0694] Experimental Example 1 TS-FeSSIF & TS-FeSSGF Solubility Test

[0695] 1. Preparation of FeSSIF

[0696] Prepare buffer B: Dissolve 4.040 g of sodium hydroxide, 8.650 g of glacial acetic acid, and 11.874 g of sodium chloride in approximately 900 ml of ultrapure water. Adjust the pH of the solution to 5.0 with 1 M sodium hydroxide or 1 M hydrochloric acid. Then dilute the solution to 1000 M with ultrapure water at room temperature.

[0697] Add powder: Add 11.200 g of FaSSIF, FeSSIF, and FaSSGF powder to approximately 500 mL of Buffer B. Stir until the powder is completely dissolved. Then dilute the solution to 1000 mL with Buffer B at room temperature.

[0698] Ready to use: Use within 48 hours at room temperature or within 24 hours at 37°C.

[0699] 2. Preparation of FeSSGF

[0700] Prepare buffer D: dissolve 1.220 g sodium acetate, 0.514 g glacial acetic acid, and 6.926 g sodium chloride in approximately 500 ml ultrapure water.

[0701] Mixed with milk: Mix with milk in an equal volume (1:1) and adjust the pH of the solution to 5.0 with 1 mol / L hydrochloric acid.

[0702] Ready to use: Use within 48 hours at room temperature or within 24 hours at 37°C.

[0703] 3. Solubility determination:

[0704] Stock solutions of test compounds and control compounds were prepared in DMSO (Solarbio S&T Co., LTD) at a concentration of 10 mmol / L.

[0705] First, 50 microliters of stock solution (10 mmol / L) of each sample was added to a vial of an uncovered solubility sample plate. The assay was performed in duplicate. DMSO was then evaporated using a centrifugal vacuum evaporator. 500 microliters of buffer were added to dissolve the samples separately. A stirring bar was placed on each vial and sealed with a PTFE / silicone stopper. The sample plate was then transferred to an Eppendorf Thermomixer Comfort plate shaker and shaken at 1100 rpm for 24 hours at 25°C. Afterwards, the stirring bar was removed using a large magnet and the sample was transferred from the solubility sample plate to a filter plate. The supernatant was centrifuged at 4,000 rpm and 25°C for 30 minutes. 350 microliters of aliquots were transferred from the supernatant. The tip was placed in acetonitrile for 5 seconds and then in water for 5 seconds. The first 25 microliters were discarded and 300 microliters were distributed to another 96-bottle glass insert plate and centrifuged again (4,000 rpm, 25°C, 30 minutes). Take a 5 μl aliquot of the supernatant and 5 μl of DMSO, then add 490 μl of a 1:1 mixture of water and acetonitrile containing an internal standard. Depending on the peak shape, dilute the solution with a certain ratio of ultrapure water. The dilution factor varies depending on the solubility value and LC-MS signal response.

[0706] Add 50 μL of stock solution (10 mmol / L) of each sample to a vial of an uncovered solubility plate. The assay was performed in duplicate. The DMSO was then evaporated using a centrifugal vacuum evaporator. 500 μL of DMSO was added to dissolve the samples. A stirring bar was placed on each vial and sealed with a molded PTFE / silicone stopper. The solubility plate was then transferred to an Eppendorf Thermomixer Comfort plate shaker and shaken at 1100 rpm for 2 hours at 25°C. At the end of 2 hours, each compound should be completely dissolved. Take a 10 μL aliquot and add 990 μL of a mixture of water and acetonitrile containing an internal standard (1:1) to 10 μL. The concentration of the standard sample may vary depending on the LC / MS signal response.

[0707] All calculations were performed using Microsoft Excel. Samples were analyzed and quantified using LC / MS / MS based on standards of known concentrations. The solubility of the test compound was calculated as follows: [Sample] = (Area ratio Sample ×DF Sample ×[STD]) / Area ratio STD , DF represents the dilution factor.

[0708] The experimental results are shown in Table 6:

[0709] Table 6 Solubility test results of the disclosed compounds

[0710] The compounds disclosed herein have good solubility, which is beneficial for in vivo absorption and subsequent formulation development.

Claims

1. A compound represented by general formula (I), a pharmaceutically acceptable salt thereof or an isomer thereof, in, X 1 Select N, or CR 5 ; X 2 Select N, or CR 6 ; X 3 Select N, or CR 7 ; X 4 Select N, or CR 8 ; X 5 Selected from N, or C; X 6 Selected from N, or C; X 7 Selected from N, or C; The condition is that when X 2 Selected from CR 6 , and X 5 Selected from C, and X 6 Select N, and X 7 When selected from C, X 3 and X 4 Not at the same time CH; R 1 and R 2 are independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy C 1-4 Alkyl or deuterated C 1-4 alkyl; R 3 Selected from -H, -CN, -OH, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclic group, 3-6 membered heterocycloalkenyl, deuterated C 1-4 alkyl, or 5-6 membered heteroaryl; wherein the phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 3a Substituted, the R 3a Selected from halogen, C 1-4 Alkyl, or C 1-4 alkoxy; L is selected from C 2-4 Alkynylidene, C 1-4 Alkylene, or C 2-4 alkenylene; R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, and 4-6 membered heterocyclic groups are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH; R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, p-methoxybenzyl, C 1-4 Alkyl-C(O)-, C 3-6 Cycloalkyl, or halogenated C 1-4 Alkyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen; R 4 Selected from -H, halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl; R 5 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl; R 6 Selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy, -COOH, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl-S(O)2-, hydroxyl C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3-6 Cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, 5-6 membered heteroaryl, or -CD3; wherein the C 1-4 Alkyl, C 3-6 Cycloalkyl, C 5-6 Cycloalkenyl, C 3-6 Cycloalkyl-C 1-4 Alkyl, C 3-6 Cycloalkyl-O-, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, phenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 6a Substituted, the R 6a Selected from halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, or C 1-4 alkoxy; R d and R e are independently selected from -H, C 1-4 Alkyl, -S(O)2-N(R f )(R g ), -C(O)-aryl, -C(O)-NR f -aryl, or -(C=S)-NR f -aryl; R f and R g are independently selected from -H, or C 1-4 alkyl; Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 cycloalkenyl; and / or, R 3 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl or 5-6 membered heteroaryl; and / or, R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 Cycloalkenyl or 5-6 membered heteroaryl; The phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkenyl and C 5-6 Cycloalkenyl is optionally further substituted by one or more alkyl radicals selected from halogen, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 The alkyl-C(O)- group is substituted.

2. The compound according to claim 1, its pharmaceutically acceptable salt or its isomer, wherein X 1 Selected from CR 5 , X 2 Selected from N, X 5 Selected from C, X 6 Selected from N, X 7 Selected from C.

3. The compound according to claim 1, its pharmaceutically acceptable salt or its isomer, wherein X 1 、X 2 All selected from N, X 5 Selected from C, X 6 Selected from N, X 7 Selected from C.

4. The compound according to any one of claims 1 to 3, its pharmaceutically acceptable salt or its isomer, wherein: R 1 、R 2 Each independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH or -CD3; Preferably, R 1 、R 2 are selected from -CH3 or -CD3.

5. The compound according to any one of claims 1 to 4, its pharmaceutically acceptable salt or its isomer, wherein: R 3 Selected from -H, -CN, -NH2, -F, -Br, -Cl, -CH3, -CH2CH3, -CH2CH(CH3)2, -CF3, -OCH3, -OCH2CH3, -CH=CH2, -N(CH3)2, -N(CH2CH3)2, -NHPMB, phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Ethylene, -C≡CR c , -OH, -CH(CH3)2, -CH2CH2CH3, -CH2CH(CH3)2, -OCH3, -CH2F, -CHF2, -CH=CH2, -CH=C(CH3)2, -CH=CHCH3, -C(CH3)=CH2, -NHCH3, -NHCH2CH3, -NHC(O)CH3, or-CD3,R c Selected from optionally one or more R ca The following groups are substituted: methyl, ethyl, cyclopropyl, cyclobutyl, isopropyl, oxetanyl, or azetidine, wherein R ca Selected from -F, -Cl, or -OH.

6. The compound according to claim 5, its pharmaceutically acceptable salt or its isomer, wherein -C≡CR c Selected from -C≡C-CH3, 7. The compound according to any one of claims 1 to 6, its pharmaceutically acceptable salt or its isomer, wherein: R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, cyclopropyl, methoxy, or -CH2CF3; Preferably, R 4 Selected from H.

8. The compound according to any one of claims 1-2 and 4-7, or a pharmaceutically acceptable salt or isomer thereof, wherein: R 5 Selected from -H, -Cl, -F, -CH3, -CN, -CF3, or -OCH3.

9. The compound according to any one of claims 1 to 8, its pharmaceutically acceptable salt or its isomer, wherein: R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -F, -Br, -Cl, -CH3, -CH2CH3, -OCH3, or -OCH2CH3; Preferably, R 7 and R 8 are independently selected from H or F.

10. The compound according to any one of claims 1 to 9, a pharmaceutically acceptable salt thereof, or an isomer thereof, which is selected from the structure represented by the following general formula: X 1 、X 2 、R 1 、R 2 、R 3 、R 4 、R 5 、R 7 and R 8 As described in any one of claims 1 to 9.

11. The compound according to any one of claims 1 and 4 to 9, its pharmaceutically acceptable salt or its isomer, wherein: X 1 Select from N;X 2 Selected from CR 6 ;X 5 Selected from C, X 6 Selected from N, X 7 Selected from C, X 3 Select N, or CR 7 , X 4 Select N, or CR 8 ; and X 3 、X 4 Not CH at the same time.

12. The compound according to claim 11, its pharmaceutically acceptable salt or its isomer, wherein R 1 、R 2 Each is independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH or -CD3; preferably, R 1 、R 2 are all selected from -CH3 or -CD3; R 3 CH3、-NH2、-OH、-F、-Cl、-Br、-CH(CH3)2、-CH2CH3、-CH2CH2CH3、-CH2CH(CH3)2、-OCH3、-CH2F、-CHF2、-CF3、-CH=CH2、-CH=C(CH3)2、-CH=CHCH3、-C(CH3)=CH2、-NHCH3、-NHCH2CH3、-NHC(O)CH3、 or -CD3; R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, cyclopropyl, or -CH2CF3; R 6 selected from -CH3, -H, -OH, -Cl, -F, -Br, -COOH, -CN, -CH2CH3, -CH(CH3)2, -CH2CH(CH3)2, -CF3, -CHF2, -CH2F, -CF2CH3, -CF(CH3)2, -OCH3, -OCH(CH3)2, -OCF3, -OCH2CF3, -N(CH3)2, -CH=CH2, -C(CH3)=CH2, -C≡CH, -S(O)2CH3, -CH2CH2CH=CH2, -CH2OH, -CH2CF3, or -CD3; X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ; R 7 and R 8 are independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3; and R 7 and R 8 Not at the same time H; Or, R 2 、R 8 The atoms connected to it together form C 5-6 Cycloalkenyl.

13. The compound according to claim 11, its pharmaceutically acceptable salt or its isomer, wherein X 3 Selected from CR 7 , X 4 Selected from CR 8 ; R 1 and R 2 Selected from C 1-4 Alkyl, or deuterated C 1-4 Alkyl; preferably, R 1 、R 2 Selected from -CH3 or -CD3; R 3 Selected from C 1-4 Alkyl; preferably, R 3 Selected from -CH3 or -CH2CH3; R 4 Selected from -H; R 6 Selected from halogen, or halogenated C 1-4 Alkyl; preferably, R 6 Selected from -Cl, -Br, or -CF3; R 7 and R 8 are independently selected from -H or halogen, and R 7 and R 8 are not hydrogen at the same time; preferably, R 7 and R 8 are independently selected from -H or F, and R 7 and R 8 Not hydrogen at the same time.

14. The compound according to claim 11, its pharmaceutically acceptable salt or its isomer, wherein R 1 and R 2 are independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH, or -CD3; preferably, R 1 and R 2 are all selected from -CH3, or -CD3; R 3 and R 6 Together with the atoms to which they are attached, they form the following groups: R 4 Selected from -H, -Cl, -F, -Br, -CH3, -CH2CH3, or cyclopropyl; X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ; R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

15. The compound according to claim 11, its pharmaceutically acceptable salt or its isomer, wherein R 1 and R 2 Each is independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH or -CD3; preferably, R 1 and R 2 are all selected from -CH3, or -CD3; R 3 is selected from -H, -CH3, or -CH2CH3; R 4 、R 6 Together with the atoms to which they are attached, they form the following groups: X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ; R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

16. The compound according to any one of claims 1 and 11-15, or a pharmaceutically acceptable salt thereof, or an isomer thereof, having any of the following structures: Ring A is selected from phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl, the phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, and 5-6 membered heteroaryl may be further substituted by one or more selected from halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 Substitution of alkyl-C(O)-; in, X 3 、X 4 、R 1 、R 2 、R 3 、R 4 、R 6 、X 3 、X 4 、R 7 and R 8 As described in any one of claims 1 and 11-15.

17. A compound represented by general formula (II), a pharmaceutically acceptable salt thereof, or an isomer thereof, X 1 Select N, or CR 5 ; X 3 Select N, or CR 7 ; X 4 Select N, or CR 8 ; R 1 and R 2 are independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy C 1-4 Alkyl or deuterated C 1-4 alkyl; R 3 Selected from -CN, -OH, -N(R a )(R b ), halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -LR c , phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclic group, 3-6 membered heterocycloalkenyl, deuterated C 1-4 alkyl, or 5-6 membered heteroaryl; wherein the phenyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkenyl, and 5-6 membered heteroaryl are optionally substituted with one or more R 3a Substituted, the R 3a Selected from halogen, C 1-4 Alkyl, or C 1-4 alkoxy; L is selected from C 2-4 Alkynylidene, C 1-4 Alkylene, or C 2-4 alkenylene; R c Selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, or 4-6 membered heterocyclic group, the C 1-4 Alkyl, C 3-6 Cycloalkyl, and 4-6 membered heterocyclic groups are optionally substituted by one or more R ca Substituted, the R ca Selected from halogen, or -OH; R a and R b are independently selected from -H, C 1-4 Alkyl, phenyl, p-methoxybenzyl, C 1-4 Alkyl-C(O)-, C 3-6 Cycloalkyl, or halogenated C 1-4 Alkyl, or R a 、R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic group, wherein the 4-6 membered heterocyclic group is optionally substituted by halogen; R 4 、R 6 Together with the atoms to which they are connected, they form a phenyl group, a 5-6 membered heterocyclic alkenyl group, a C 5-6 cycloalkenyl, or 5-6 membered heteroaryl; the phenyl, 5-6 membered heteroaryl, 5-6 membered heterocycloalkenyl, and C 5-6 Cycloalkenyl is optionally further substituted by one or more alkyl radicals selected from halogen, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 substituted by an alkyl-C(O)- substituent; R 5 、R 7 and R 8 are independently selected from -H, -OH, -CN, -N(R d )(R e ), halogen, C 1-4 Alkyl, C 1-4 Alkoxy or halogenated C 1-4 alkyl.

18. The compound according to claim 17, its pharmaceutically acceptable salt or its isomer, wherein R 1 and R 2 Each independently selected from -CH3, -CH2CH3, -CH(CH3)2, -Cl, -F, -Br, -OCH3, -CH2OH, or -CD3; R 3 For -CH3, -CH2CH3; R 4 、R 6 Together with the atoms to which they are attached, they form the following groups: R 5 Selected from -H, -CN, halogenated C 1-3 Alkyl or halogen; R 7 and R 8 Each is independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -Br, -OCH3, or -OCH2CH3.

19. The compound according to claim 17 or 18, or a pharmaceutically acceptable salt thereof or an isomer thereof, wherein: X 1 Selected from CR 5 ;X 3 Selected from CR 7 ;X 4 Selected from CR 8 ; R 1 and R 2 are independently selected from C 1-4 Alkyl, preferably -CH3; R 3 Selected from C 1-4 Alkyl, preferably -CH3; R 4 、R 6 Together with the atoms to which it is connected, it forms a 5-6 membered heteroaryl group, preferably a pyridyl group, more preferably R 5 is selected from -H or halogen, preferably H or F, more preferably H; R 7 and R 8 are each independently selected from -H.

20. The compound according to any one of claims 17 to 19, or a pharmaceutically acceptable salt thereof or an isomer thereof, having any of the following structures: Ring B is independently selected from phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, or 5-6 membered heteroaryl, the phenyl, 5-6 membered heterocycloalkenyl, C 5-6 Cycloalkenyl, and 5-6 membered heteroaryl are optionally further substituted by one or more selected from halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, hydroxyl C 1-6 Alkyl, and C 1-6 Substitution of alkoxy groups; in, R 3 、R 4 、R 5 、R 6 、X 3 、X 4 、R 7 and R 8 As described in any one of claims 17 to 19.

21. The compound according to any one of claims 1 and 4 to 9, its pharmaceutically acceptable salt or its isomer, wherein: X 1 Selected from N, X 5 Selected from N, X 2 Selected from CR 6 , X 6 Selected from C, X 7 Selected from C.

22. The compound according to any one of claims 1 and 4 to 9, its pharmaceutically acceptable salt or its isomer, wherein: X 1 Selected from N, X 2 Selected from CR 6 , X 5 Selected from C, X 6 Selected from C, X 7 Selected from N.

23. The compound according to claim 21 or 22, its pharmaceutically acceptable salt or its isomer, wherein R 1 and R 2 are independently selected from -CH3, -CH2CH3, or -CH(CH3)2; preferably, R 1 and R 2 are all selected from -CH3; R 3 Selected from -CH3; R 4 Selected from -H; R 6 Selected from -CH3; X 3 Select N, or CR 7 ;X 4 Select N, or CR 8 ; R 7 and R 8 Each independently selected from -H, -OH, -CN, -NH2, -CH3, -CH2CH3, -Cl, -F, -OCH3, or -OCH2CH3; Or, R 2 、R 8 Together with the atoms it is connected to form C 5-6 Cycloalkenyl.

24. The compound according to any one of claims 21 to 23, a pharmaceutically acceptable salt thereof, or an isomer thereof, having any of the following structures: in, R 1 、R 2 、R 3 、R 4 、R 6 、X 3 、X 4 As described in any one of claims 21 to 23.

25. [Corrected 02.02.2024 in accordance with Rule 91] A compound as described below, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein the compound is selected from any of the following structures:

26. A pharmaceutical composition comprising the compound according to any one of claims 1 to 25, a pharmaceutically acceptable salt or an isomer thereof, and one or more pharmaceutically acceptable carriers.

27. Use of the compound according to any one of claims 1 to 25, a pharmaceutically acceptable salt thereof, or an isomer thereof in the preparation of a medicament for preventing and / or treating a PKMYT1-mediated tumor disease; the PKMYT1-mediated tumor disease is preferably a PKMYT1-mediated tumor disease caused by CCNE1 overexpression and / or FBXW7 inactivation mutation.

28. A method for treating a tumor disease mediated by PKMYT1, comprising administering to a subject a therapeutically or prophylactically effective amount of a compound according to any one of claims 1 to 25, a pharmaceutically acceptable salt thereof, or an isomer thereof; the PKMYT1-mediated tumor disease is preferably a PKMYT1-mediated tumor disease caused by CCNE1 overexpression and / or FBXW7 inactivation mutation.

29. The use according to claim 27 or the method according to claim 28, wherein the neoplastic disease is selected from one or more of ovarian cancer, breast cancer, gastric cancer, esophageal cancer, lung cancer, endometrial cancer, and colorectal cancer.

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