Azaquinazoline ring derivative and use thereof

Abstract

The present invention provides an azaquinazoline ring derivative and a use thereof. Specifically, the present invention provides a compound of formula I, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotopic substitute thereof. The compound has excellent cancer Osaka thyroid (Cot) kinase inhibitory activity, and thus can be used for preventing or treating Cot kinase-mediated diseases or disorders.

Description

Azaquinazoline ring derivatives and uses thereof Technical Field

[0001] The present invention relates to a series of azaquinazoline ring derivatives and uses thereof, and particularly to compounds represented by formula I and pharmaceutically acceptable salts thereof. Background Art

[0002] The Cot (Osaka thyroid carcinoma) protein is a serine / threonine kinase that is a member of the MAP kinase kinase kinase (MAP3K) family. It is also known as "TPL2" (tumor progression locus), "MAP3K8" (mitogen-activated protein kinase kinase kinase 8), or "EST" (Ewing sarcoma transformant). Cot was originally discovered as an oncogene in human thyroid cancer cell lines and has since been shown to play an important role in regulating inflammatory pathways.

[0003] Cot is located downstream of receptors such as TNF-αR, IL-1R, and TLRs, promoting the production and signaling of inflammatory factors such as TNF-α and IL-1β by activating the MEK-ERK pathway. TNF-α is a proinflammatory cytokine and plays a key role in immune-related diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), inflammatory bowel disease (IBD), diabetes, sepsis, psoriasis, and transplant rejection. Cot can also promote the secretion of inflammatory mediators by neutrophils by activating p38α and p38δ.

[0004] Therefore, there is an urgent need in the art to provide drugs with better activity, better metabolic stability and better safety that can treat diseases related to Cot activity.

[0005] Summary of the Invention

[0006] The present invention aims to provide a class of Osaka thyroid carcinoma (Cot) kinase inhibitor compounds with novel structure and excellent inhibitory activity, and also provides the use thereof in treating diseases or conditions mediated by Osaka thyroid carcinoma (Cot) kinase.

[0007] In a first aspect, the present invention provides a compound of formula I, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substitution thereof,

[0008] in,

[0009] X 1 N or CR 15 ;

[0010] X 2 and X 3 are independently N or CH;

[0011] where X1 、X 2 and X 3 At least one is N;

[0012] R 1 For hydrogen, C 1-9 Alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 1 ;

[0013] R 2 is hydrogen, -C(O)-R 7 、-C(O)OR 7 、-C(O)N(R 7 )2、C 1-9 Alkyl, C 3-15 Cycloalkyl;

[0014] R 3 is a heterocyclic group or a heteroaryl group, wherein each heterocyclic group or heteroaryl group optionally has 1 to 4 Z 3 ;

[0015] R 4 is aryl, heterocyclic, or heteroaryl, wherein the aryl, heterocyclic, or heteroaryl is optionally substituted with 1 to 4 Z 4 ;

[0016] R 5 For hydrogen, halogen, -CN, -NO2, -OR 7 、-N(R 8 )(R 9 )、-S(O)-R 7 、-S(O)2R 7 、-S(O)2N(R 7 )2、-C(O)R 7 、-OC(O)-R 7 、-C(O)OR 7 、-OC(O)OR 7 、-OC(O)N(R 10 )(R 11 )、-C(O)N(R 7 )2、-N(R 7 )C(O)(R 7 ), C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-9 Alkylthio, C 1-6 Halogenated alkyl, C 3-15Cycloalkyl, aryl, heterocyclyl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-9 Alkylthio, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl and heteroaryl groups may be optionally substituted with 1 to 4 Z 5 ;

[0017] R 6 is hydrogen, -C(O)-R 7 、-C(O)OR 7 、-C(O)N(R 7 )2、C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 3-10 Heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 6 ;

[0018] Each R 7 are independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl, or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl and heteroaryl groups may be optionally substituted with 1 to 4 Z 7 ; R 8 and R 9 is independently hydrogen at each occurrence, -S(O)2-R 10 、-C(O)-R 10 、-C(O)OR 10 、-C(O)N(R 10 )(R 11) 、C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 8 ;

[0019] R 10 and R 11 is independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 9 ;

[0020] Every Z 1 、Z 2 、Z 3 、Z 4 、Z 5 、Z 6 、Z 7 、Z 8 and Z 9 are independently hydrogen, oxo, halogen, -NO2, -N3, -CN, thio, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-8 Halogenated alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl, -OR 12 、-C(O)R 12 、-C(O)OR 12 、-C(O)N(R 13 )(R 14 )、-N(R 13 )(R 14 )、-N(R 13 )2(R 14 ) + 、-N(R 12 )C(O)-R 12 、-N(R 12 )C(O)OR 12 、-N(R12 )C(O)N(R 13 )(R 14 )、-N(R 12 )S(O)2(R 12 )、-NR 12 S(O)2N(R 13 )(R 14 )、-NR 12 S(O)2O(R 12 )、-OC(O)R 12 、-OC(O)-N(R 13 )(R 14 )、-P(O)(OR 12 )2、-OP(O)(OR 12 )2、-CH2P(O)(OR 12 )2、-OCH2P(O)(OR 12 )2、-C(O)OCH2P(O)(OR 12 )2、-P(O)(R 12 )-(OR 12 )、-OP(O)(R 12 )(OR 12 )、-CH2P(O)(R 12 )(OR 12 )、-OCH2P(O)(R 12 )(OR 12 )、-C(O)OCH2P(O)-(R 12 )(OR 12 )、-P(O)(N(R 12 )2)2、-OP(O)(N(R 12 )2)2、-CH2P(O)(N(R 12 )2)2、-OCH2P(O)(N(R 12 )2)2、-C(O)OCH2P(O)(N(R 12 )2)2、-P(O)(N(R 12 )2)(OR 12 )、-OP(O)(N(R 12 )2)(OR 12 )、-CH2P(O)(N(R 12 )2)-(OR 12 )、-OCH2P(O)(N(R 12 )2)(OR 12 )、-C(O)OCH2P(O)(N(R 12 )2)(OR 12 )、-P(O)(R 12)(N(R 12 )2)、-OP(O)(R 12 )(N(R 12 )2)、-CH2P(O)(R 12 )(N(R 12 )2)、-OCH2P(O)(R 12 )(N(R 12 )2)、-C(O)OCH2P(O)-(R 12 )(N(R 12 )2),-SR 12 、-S(O)R 12 、-S(O)(NH)R 12 、-S(O)2R 12 Or -S(O)2N(R 13 )(R 14 ); wherein any alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, aryl, heteroaryl or heterocyclic group is optionally substituted by 1 to 4 substituents selected from the group consisting of oxo, halogen, thio, hydroxy, -NO2, -CN, -N3, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 1-9 Alkoxy, C 1-9 Alkyl hydroxyl, -COOH, -COOC 1-9 Alkyl, amino, C 1-9 Alkylamino, (C 1-9 Alkyl)2amino, C 1-8 Halogenated alkyl, C 1-9 Alkylcarbonyl, -CONH2, -CON(C 1-9 alkyl) 1-2 、C 1-9 Alkylsulfonyl, aryl, heteroaryl, heterocyclic;

[0021] Each R 12 are independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, aryl, heterocyclic, or heteroaryl, wherein any alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, or heterocyclic group optionally has 1 to 4 Z 7 group;

[0022] R 13 and R 14 Each occurrence is independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C3-15 Cycloalkyl, heterocyclic, aryl or heteroaryl; wherein any alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl or heterocyclic group optionally has 1 to 4 Z 8 group; or R 13 and R 14 Together with the nitrogen to which they are attached, they form a heterocyclic group, wherein the heterocyclic group is optionally substituted with 1 to 4 Z 8 group;

[0023] Each R 15 are independently H, halogen, -CN, -NO2, -OR 7 、-N(R 8 )(R 9 )、-S(O)-R 7 、-S(O)2R 7 、-S(O)2N(R 7 )2、-C(O)R 7 、-OC(O)-R 7 、-C(O)OR 7 、-OC(O)OR 7 、-OC(O)N(R 10 )(R 11 )、-C(O)N(R 7 )2、-N(R 7 )C(O)(R 7 ), C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-9 Alkylthio, C 1-6 Halogenated alkyl, C 3-15 cycloalkyl, aryl, heterocyclyl or heteroaryl.

[0024] Every Z 1 、Z 2 、Z 3 、Z 4 、Z 5 、Z 6 、Z 7 、Z 8 and Z 9 are independently hydrogen, oxo, halogen, -NO2, -N3, -CN, thio, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-8 Halogenated alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl, -OR 12 、-C(O)R 12 、-C(O)OR 12、-C(O)N(R 13 )(R 14 )、-N(R 13 )(R 14 )、-N(R 13 )2(R 14 ) + 、-N(R 12 )C(O)-R 12 、-N(R 12 )C(O)O-R 12 、-N(R 12 )C(O)N(R 13 )(R 14 )、-N(R 12 )S(O)2(R 12 )、-NR 12 S(O)2N(R 13 )(R 14 )、-NR 12 S(O)2O(R 12 )、-OC(O)R 12 、-OC(O)-N(R 13 )(R 14 )、-P(O)(OR 12 )2、-OP(O)(OR 12 )2、-CH2P(O)(OR 12 )2、-OCH2P(O)(OR 12 )2、-C(O)OCH2P(O)(OR 12 )2、-P(O)(R 12 )-(OR 12 )、-OP(O)(R 12 )(OR 12 )、-CH2P(O)(R 12 )(OR 12 )、-OCH2P(O)(R 12 )(OR 12 )、-C(O)OCH2P(O)-(R 12 )(OR 12 )、-P(O)(N(R 12 )2)2、-OP(O)(N(R 12 )2)2、-CH2P(O)(N(R 12 )2)2、-OCH2P(O)(N(R 12 )2)2、-C(O)OCH2P(O)(N(R 12 )2)2、-P(O)(N(R 12 )2)(OR 12)、-OP(O)(N(R 12 )2)(OR 12 )、-CH2P(O)(N(R 12 )2)-(OR 12 )、-OCH2P(O)(N(R 12 )2)(OR 12 )、-C(O)OCH2P(O)(N(R 12 )2)(OR 12 )、-P(O)(R 12 )(N(R 12 )2)、-OP(O)(R 12 )(N(R 12 )2)、-CH2P(O)(R 12 )(N(R 12 )2)、-OCH2P(O)(R 12 )(N(R 12 )2)、-C(O)OCH2P(O)-(R 12 )(N(R 12 )2),-SR 12 、-S(O)R 12 、-S(O)(NH)R 12 、-S(O)2R 12 Or -S(O)2N(R 13 )(R 14 ); wherein any alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, aryl, heteroaryl or heterocyclic group is optionally substituted by 1 to 4 substituents selected from the group consisting of oxo, halogen, thio, -NO2, -CN, -N3, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 1-9 Alkoxy, amino, C 1-9 Alkylamino, (C 1-9 Alkyl)2amino, C 1-8 Halogenated alkyl, C 1-9 Alkylcarbonyl, C 1-9 Alkylsulfonyl, aryl, heteroaryl, heterocyclic.

[0025] In another preferred embodiment, the compound of formula I has the structure shown in formula Ia:

[0026] where X 1 、X 2 、X 3 、R 1 -R 5 and R 6As described in this article.

[0027] In another preferred embodiment, the compound of formula I has the structure shown in formula Ib:

[0028] where X 1 、X 2 、X 3 、R 1 -R 5 and R 6 As described in this article.

[0029] In another preferred embodiment, the compound of formula I is a mixture of formula Ia and formula Ib. Preferably, the proportion of formula Ia in the mixture is >50%, more preferably ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, or even ≥98% or ≥99%.

[0030] In another preferred embodiment, wherein R 1 and R 2 There is only one hydrogen in it.

[0031] In another preferred embodiment, the compound has a structure shown in Formula II:

[0032] where X 1 、X 2 、X 3 、R 1 、R 3 、R 4 、R 5 and R 6 As defined herein.

[0033] In another preferred embodiment, the compound has the structure shown in Formula IIa:

[0034] where X 1 、X 2 、X 3 、R 1 、R 3 、R 4 、R 5 and R 6 As defined herein.

[0035] In another preferred embodiment, R 2 For hydrogen.

[0036] In another preferred embodiment, R 1 Selected from C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 3-10 Heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 substituents independently selected from the group consisting of halogen, -OH, -CN, -OR 12 、C 1-9 Alkyl and aryl groups.

[0037] In another preferred embodiment, R 1 Selected from C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Cycloalkyl, 3-7 membered heterocyclyl, C6 aryl or 5-7 membered heteroaryl, wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Cycloalkyl, 3-7 membered heterocyclyl, C6 aryl or 5-7 membered heteroaryl may be optionally substituted with 1 to 4 substituents independently selected from the group consisting of halogen, -OH, -CN, -OR 12 、C 1-9 Alkyl and C6 aryl.

[0038] In another preferred embodiment, R 1 Select from the following groups:

[0039] In another preferred embodiment, X 1 、X 2 and X 3 are independently N or CH, wherein X 1 、X 2 and X 3 At least one of them is N.

[0040] In another preferred embodiment, R 15 Each occurrence is independently H, CN, halogen, OH, C 1-9 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl.

[0041] In another preferred embodiment, X 1 、X 2 and X 3 There is only one N or X 1 、X 2 and X 3 There are two N.

[0042] In another preferred embodiment, X 1 、X 2 and X 3 There is only X 3 is N.

[0043] In another preferred embodiment, the compound has a structure shown in Formula III-1, III-2 or III-3:

[0044] Among them, each R 1 、R 3 、R 4 、R 5 and R 6 Independently as described herein.

[0045] In some embodiments, the compound of Formula I is represented by Formula III-1a, III-2a, and III-3a:

[0046] where R 1 、R 3 、R 4 、R 5 and R 6 Independently as described herein.

[0047] In another preferred embodiment, R 5 Selected from hydrogen, halogen, -CN, -OR 7 、-S(O)-R 7 、-S(O)2R 7 、-S(O)2N(R 7 )2、-C(O)R 7 、-C(O)N(R 7 )2、C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, aryl, heterocyclyl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, aryl, heterocyclyl or heteroaryl may be optionally substituted with 1 to 4 substituents independently selected from the group consisting of halogen, -CN, -OR 12 、C 1-9 Alkyl and aryl groups.

[0048] In another preferred embodiment, R 5 is selected from F, Cl, Br or CN.

[0049] In another preferred embodiment, R 6is selected from hydrogen, methyl, ethyl or cyclopropyl.

[0050] In another preferred embodiment, R 6 For hydrogen.

[0051] In another preferred embodiment, R 3 is a heteroaryl group, wherein each heteroaryl group optionally has 1 to 4 Z 3 .

[0052] In another preferred embodiment, R 3 Select from the following groups:

[0053] In another preferred embodiment, the compound has a structure shown in Formula IV-1, IV-2 or IV-3:

[0054] Among them, each Z 3 、R 1 、R 4 、R 5 and R 6 Independently as described herein.

[0055] In another preferred embodiment, the compound has a structure shown in formula IV-1a, IV-2a or IV-3a

[0056] where Z 3 、R 1 、R 4 、R 5 and R 6 Independently as defined herein.

[0057] In another preferred embodiment, Z 3 C 1-9 Alkyl, C 3-15 Cycloalkyl and heterocyclic groups; any C 1-9 Alkyl, C 3-15 Cycloalkyl and heterocyclyl are optionally selected from hydrogen, oxo, halogen, hydroxy, thio, -NO2, -CN, -N3, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 1-9 Alkoxy, C 1-9 Alkyl hydroxyl, -COOH, -COOC 1-9 Alkyl, amino, C 1-9 Alkylamino, (C 1-9 Alkyl)2amino, C 1-9 alkyl, C 1-9 Alkylcarbonyl, -CONH2, -CON(C 1-9 alkyl)1-2 、C 1-9 The alkylsulfonyl group is substituted by 1 to 4 substituents selected from the group consisting of alkylsulfonyl, aryl, heteroaryl, and heterocyclic groups.

[0058] In another preferred embodiment, Z 3 Selected from

[0059] In another preferred embodiment, Z 3 Selected from

[0060] In another preferred embodiment, Z 3 Selected from

[0061] In another preferred embodiment, R 3 Selected from

[0062] In another preferred embodiment, R 4 is a heterocyclic or heteroaryl group, and the heterocyclic or heteroaryl group is optionally substituted with 1 to 3 substituents independently selected from the following: -CN, halogen, -OR 12 、-C(O)-R 12 、C 1-9 Alkyl, C 1-9 Haloalkyl and heterocyclic groups.

[0063] In another preferred embodiment, R 4 is: pyrimidinyl, pyridinyl, pyrazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, isoquinolinonyl, phthalazinonyl, indazolyl, benzopyrimidinyl, pyridopyridinyl and imidazopyridinyl;

[0064] The H atoms in the structure are optionally substituted by substituents selected from the group consisting of: -CN, halogen, -OR 12 、-C(O)-R 12 、C 1-9 Alkyl, C 1-9 Haloalkyl and heterocyclic groups.

[0065] In another preferred embodiment, R 4 Selected from

[0066] Furthermore, R 4 Selected from

[0067] In another preferred embodiment, the compound has a structure of formula IV-1 or IV-1a, wherein Z 3 Selected from R 5 is halogen or cyano;

[0068] R 1 Select from the following groups: R 4 Select from the following groups: and R 6 is selected from hydrogen, methyl, ethyl or cyclopropyl.

[0069] In another preferred embodiment, X 1 and X 2 CH, X 3 is N, and R 5 For F, Cl and CN.

[0070] In another preferred embodiment, each X 1 、X 2 、X 3 、R 1 -R 6 and R 15 are independently the corresponding groups in the compounds described in the following table.

[0071] In another preferred embodiment, the compound is selected from:

[0072] In another preferred embodiment, the isotope substitute is a deuterated substitute.

[0073] In a second aspect, the present invention provides a pharmaceutical composition.

[0074] It comprises the compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substitution thereof, and a pharmaceutically acceptable carrier.

[0075] In another preferred embodiment, the pharmaceutical composition further comprises another therapeutic agent, such as an anti-inflammatory agent or an anti-cancer agent.

[0076] In a third aspect, the present invention provides the use of the compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substitution thereof, or the pharmaceutical composition described in the second aspect of the present invention in the preparation of a medicament for preventing or treating diseases or conditions mediated by Osaka thyroid carcinoma (Cot) kinase.

[0077] In another preferred embodiment, the disease or condition is selected from the following group: cancer, diabetes, and inflammatory diseases.

[0078] In another preferred embodiment, the disease or condition is selected from the group consisting of type 1 and type 2 diabetes, non-alcoholic fatty liver disease (NAFLD) and inflammatory diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), inflammatory bowel disease (IBD), sepsis, psoriasis, dysregulated TNF expression and transplant rejection; acute gout and ankylosing spondylitis, allergic diseases, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis and multiple sclerosis, atherosclerosis, bacterial infection, bone cancer pain and endometriosis. pain; BRAF-resistant melanoma, colon cancer, brainstem glioma or pituitary adenoma, burns, bursitis, cancer of the anal area, cancer of the endocrine system, cancer of the kidney or ureter (e.g., renal cell carcinoma, renal pelvis cancer), cancer of the penis, cancer of the small intestine, non-small cell lung cancer, thyroid cancer, cancer of the urethra, blood cancers such as acute myeloid leukemia, tongue cancer, cervical cancer, endometrial cancer, fallopian tube cancer, renal pelvis cancer, vaginal cancer, or vulvar cancer, chronic myeloid leukemia, chronic or acute leukemia, chronic pain, classic Bartter syndrome, common cold, conjunctivitis, coronary heart disease, skin or intraocular melanoma, dermatitis, dysmenorrhea, eczema, endometriosis, familial adenomatous polyposis, fibromyalgia, fungal infection, gout, Gynecological cancer, uterine sarcoma, fallopian tube cancer, headache, hemophilic arthropathy, Parkinson's disease, AIDS, herpes zoster, Hodgkin's disease, Huntington's disease, hyperprostaglandin E syndrome, influenza, iritis, juvenile arthritis, juvenile onset rheumatoid arthritis, juvenile rheumatoid arthritis, low back and neck pain, lymphocytic lymphoma, myofascial disorders, myositis, neuralgia, neurodegenerative disorders such as Alzheimer's disease, neuritis, neuropathic pain, vulvar cancer, Parkinson's disease, Johnson's disease, pediatric malignancies, pulmonary fibrosis, colorectal cancer, rhinitis, sarcoidosis, soft tissue sarcomas, scleritis, skin cancer, solid tumors in children, spinal cord tumors, sprains and strains, gastric cancer, stroke, subacute and chronic musculoskeletal pain syndromes such as bursitis, surgical or dental procedures, symptoms associated with influenza or other viral infections, synovitis, toothache, ulcers, uterine cancer, uterine sarcoma, uveitis, vasculitis, viral infections, viral infections (e.g., influenza), and wound healing.

[0079] In a fourth aspect, the present invention provides use of the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope thereof in the preparation of an Osaka thyroid carcinoma (Cot) kinase inhibitor.

[0080] In a fifth aspect, the present invention provides an intermediate compound represented by formula M1:

[0081] Among them, R 16 is Cl, Br, I, OTf or NH2;

[0082] And X 1 、X 2 、X 3 、R 1 、R 2 and R 5 As defined in the first aspect of the present invention.

[0083] In another preferred embodiment, the intermediate compound has a structure shown in formula M1a:

[0084] Among them, R 16 is Cl, Br, I, OTf or NH2;

[0085] And R 1 、R 2 and R 5 As defined in the first aspect of the present invention.

[0086] In the sixth aspect of the present invention, a method for inhibiting Cot activity is provided, which comprises contacting a cell with at least one compound according to the first aspect of the present invention or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substitute thereof or the pharmaceutical composition, wherein the contact is in vitro, ex vivo or in vivo.

[0087] In the seventh aspect of the present invention, a method for treating a disease or condition mediated by Osaka thyroid carcinoma (Cot) kinase in a human patient in need thereof is provided, comprising the steps of administering to the patient a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers or isotope thereof, or a composition as described in the second aspect of the present invention.

[0088] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0089] After extensive and in-depth research, numerous screenings and tests, the present inventors have provided a class of novel structurally and highly effective Osaka thyroid carcinoma (Cot) kinase inhibitor compounds, on the basis of which the present invention was completed.

[0090] the term

[0091] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0092] As used herein, the terms "comprising" or "including" may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0093] The prefix "C u-v " indicates that the following group has from u to v carbon atoms. For example, "C 1-9 "Alkyl" means an alkyl group having 1 to 9 carbon atoms. u-v "Includes all positive integers of C atoms between u and v, such as 1, 2, 3, 4, 5, 6, 7, 8, and 9 carbon atoms.

[0094] "Alkyl" refers to an unbranched or branched saturated hydrocarbon chain. As used herein, an alkyl group or a portion thereof having 1 to 20 carbon atoms (i.e., C 1-20 alkyl), 1 to 9 carbon atoms (i.e., C 1-9 alkyl), 1 to 6 carbon atoms (i.e., C 1-6 alkyl), or 1 to 4 carbon atoms (i.e., C 1-4 Alkyl). Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having a specific number of carbon atoms is named by chemical name or determined by molecular formula, all isomers having that number of carbon atoms are included; thus, for example, "butyl" includes n-butyl (i.e., -(CH2)3CH3), sec-butyl (i.e., -CH(CH3)CH2CH3), isobutyl (i.e., -CH2CH(CH3)2), and tert-butyl (i.e., -C(CH3)3); and "propyl" includes n-propyl (i.e., -(CH2)2CH3) and isopropyl (i.e., -CH(CH3)2).

[0095] "Alkenyl" refers to an alkyl group containing at least one carbon-carbon double bond and having from 2 to 20 carbon atoms (i.e., C 2-20alkenyl), 2 to 8 carbon atoms (i.e., C 2-8 alkenyl), 2 to 6 carbon atoms (i.e., C 2-6 alkenyl), or 2 to 4 carbon atoms (ie, C 2-4 Examples of the alkenyl group include ethenyl, propenyl, butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

[0096] "Alkynyl" refers to an alkyl group containing at least one carbon-carbon triple bond and having from 2 to 20 carbon atoms (i.e., C 2-20 Alkynyl), 2 to 8 carbon atoms (i.e., C 2-8 Alkynyl), 2 to 6 carbon atoms (i.e., C 2-6 Alkynyl), or 2 to 4 carbon atoms (i.e., C 2-4 The term "alkynyl" also includes those groups having one triple bond and one double bond.

[0097] "Alkoxy" refers to the group "alkyl-O-," where alkyl is as defined above. Examples of alkoxy include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2-dimethylbutoxy.

[0098] "Haloalkoxy" refers to an alkoxy group as defined above wherein one or more hydrogen atoms are replaced by a halogen.

[0099] "Alkylthio" refers to the group "alkyl-S-."

[0100] "Acyl" refers to the group -C(O)R, where R is hydrogen, alkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted as defined herein. Examples of acyl include formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethyl-carbonyl, and benzoyl.

[0101] "Acylamino" refers to a "C-acylamino" group (which refers to the group -C(O)NR y R z ) and "N-amido" groups (which refer to the group -NR y C(O)R z ), where R y and R z are independently selected from hydrogen, alkyl, aryl, haloalkyl, or heteroaryl; each of which may be optionally substituted.

[0102] "Amino" refers to the group -NR y R z , where R y and R zare independently selected from hydrogen, alkyl, haloalkyl, aryl, or heteroaryl; each of which may be optionally substituted.

[0103] "Amidino" refers to -C(NH)(NH2).

[0104] "Aryl" refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic) (including fused systems). As used herein, an aryl group has 6 to 20 ring carbon atoms (i.e., C 6-20 aryl), 6 to 12 carbon ring atoms (i.e., C 6-12 aryl), or 6 to 10 carbon ring atoms (i.e., C 6-10 Examples of aryl groups include phenyl, naphthyl, fluorenyl, and anthracenyl. However, aryl groups do not include or overlap in any way with heteroaryl groups as defined below. If one or more aryl groups are fused to a heteroaryl group, the resulting ring system is a heteroaryl group. If one or more aryl groups are fused to a heterocyclyl group, the resulting ring system is a heterocyclyl group.

[0105] "Azide" refers to -N3.

[0106] "Carbamoyl" refers to an "O-carbamoyl" group (which refers to the group -OC(O)NR y R z ) and "N-carbamoyl" groups (which refer to the group -NR y C(O)OR z ), where R y and R z are independently selected from hydrogen, alkyl, aryl, haloalkyl, or heteroaryl; each of which may be optionally substituted.

[0107] "Carboxyl" refers to -C(O)OH.

[0108] "Carboxyl ester" refers to -OC(O)R and -C(O)OR, where R is hydrogen, alkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted as defined herein.

[0109] "Cyano" or "nitrile" refers to the group -CN.

[0110] "Cycloalkyl" refers to a saturated or partially unsaturated cyclic alkyl group having a single or multiple rings (including fused, bridged, and spirocyclic ring systems). The term "cycloalkyl" includes cycloalkenyl groups (i.e., the cyclic group has at least one double bond). Cycloalkyl groups, as used herein, have from 3 to 20 ring carbon atoms (i.e., C 3-20 cycloalkyl), 3 to 15 ring carbon atoms (i.e., C 3-15 cycloalkyl), 3 to 12 ring carbon atoms (i.e., C3-12 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C 3-10 cycloalkyl), 3 to 8 ring carbon atoms (i.e., C 3-8 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C 3-6 Examples of the cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0111] "Guanidino" refers to -NHC(NH)(NH2).

[0112] "Hydrazino" refers to -NHNH2.

[0113] "Imino" refers to the group -C(NR)R, where each R is alkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted as defined herein.

[0114] "Halogen" or "halogenated" includes fluorine, chlorine, bromine, and iodine.

[0115] "Haloalkyl" refers to an unbranched or branched alkyl group as defined above in which one or more hydrogen atoms are replaced by a halogen. For example, when a residue is substituted with more than one halogen, it may be referred to by using a prefix corresponding to the number of halogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkyl groups substituted with two ("di") or three ("tri") halogen groups, which may be, but need not be, the same halogen. Examples of haloalkyl groups include difluoromethyl (-CHF2) and trifluoromethyl (-CF3).

[0116] "Heteroalkyl" refers to an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by the same or different heteroatom groups. The term "heteroalkyl" includes unbranched or branched saturated chains having carbon and heteroatoms. By way of example, 1, 2 or 3 carbon atoms can be independently replaced by the same or different heteroatom groups. Heteroatom groups include, but are not limited to, -NR-, -O-, -S-, -S(O)-, -S(O)2-, etc., where R is H, alkyl, aryl, cycloalkyl, heteroalkyl, heteroaryl or heterocyclic radical, each of which can be optionally substituted. Examples of heteroalkyl groups include -OCH3, -CHOCH3, -SCH3, -CH2SCH3, -NRCH3 and -CH2NRCH3, where R is hydrogen, alkyl, aryl, arylalkyl, heteroalkyl or heteroaryl, each of which can be optionally substituted. As used herein, a heteroalkyl group can contain 1-10 carbon atoms, 1-8 carbon atoms, or 1-4 carbon atoms; and 1-3 heteroatoms, 1-2 heteroatoms, or 1 heteroatom.

[0117] "Heteroaryl" refers to an aromatic group having a monocyclic, multiple or multiple fused rings, wherein one or more ring heteroatoms are independently selected from nitrogen, oxygen and sulfur. As described herein, a heteroaryl group may include 5 to 20 ring atoms (i.e., 5-20 membered heteroaryl), 5 to 12 ring atoms (i.e., 5-12 membered heteroaryl), or 6 to 10 ring atoms (i.e., 6-10 heteroaryl); wherein, 1 to 5 heteroatoms, 1 to 4 heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom, wherein the ring heteroatoms are independently selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include pyrimidinyl, purinyl, pyridinyl, pyridazinyl, benzothiazolyl and pyrazolyl. Examples of fused heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolyl, isoquinolyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[1,5-a]pyridinyl, and imidazo[1,5-a]pyridinyl, wherein the heteroaryl group may be attached through any ring of the fused system. Any aromatic ring with single or multiple fused rings containing at least one heteroatom is considered a heteroaryl group, regardless of attachment to the rest of the molecule (i.e., through any of the fused rings). Heteroaryl groups do not include or overlap with aryl groups as defined above.

[0118] "Heterocyclic radical" refers to a saturated or unsaturated cycloalkyl radical having one or more ring heteroatoms independently selected from nitrogen, oxygen and sulphur. The term "heterocyclic radical" includes heterocyclenyl (i.e., a heterocyclic radical having at least one double bond), bridged heterocyclic radical, fused heterocyclic radical and spiral-heterocyclic radical. A heterocyclic radical can be a monocyclic or polycyclic ring, wherein the polycyclic ring can be fused, bridged or spirocyclic. Any non-aromatic ring containing at least one heteroatom is considered to be a heterocyclic radical, regardless of connection (i.e., it can be combined by carbon atoms or heteroatoms). In addition, the term heterocyclic radical is intended to include any non-aromatic ring containing at least one heteroatom, which ring can be fused to an aryl or heteroaryl ring, regardless of connection to the rest of the molecule. The heterocyclic group used in this article can have 3 to 20 ring atoms (i.e., 3-20 membered heterocyclic groups), 4 to 12 ring atoms (i.e., 4-12 membered heterocyclic groups), 5 to 10 ring atoms (i.e., 5-10 membered heterocyclic groups), 6 to 7 ring atoms (i.e., 6-8 membered heterocyclic groups), or 3 to 6 ring atoms (i.e., 3-6 membered heterocyclic groups); wherein, there can be 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom, the ring heteroatoms being independently selected from nitrogen, sulfur or oxygen. The heterocyclic group may include one or more oxo and / or thio groups. Examples of heterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl, oxetanyl, dioxolane, azetidinyl, and morpholinyl. As used herein, the term "bridged-heterocyclic radical" refers to a 4 to 10 ring moiety that is connected to a 4 to 10 ring moiety having at least one heteroatom at two non-adjacent atoms of a heterocyclic radical with one or more (e.g., 1 or 2), wherein each heteroatom is independently selected from nitrogen, oxygen, and sulphur."bridged-heterocyclic radical" as used herein includes bicyclic and tricyclic ring systems. In addition, as used herein, the term "spiro-heterocyclic radical" refers to a ring system in which a 3- to 10-membered heterocyclic radical has one or more other rings, wherein the one or more other rings are 3- to 10-membered cycloalkyl or 3- to 10-membered heterocyclic radicals, wherein the single atom of the one or more other rings is also an atom of the 3- to 10-membered heterocyclic radical. Examples of spiro-heterocyclyl rings include bicyclic and tricyclic ring systems, such as 2-oxa-7-azaspiro[3.5]nonyl, 2-oxa-6-azaspiro[3.4]octyl, and 6-oxa-1-azaspiro[3.3]heptyl. Examples of fused heterocyclyl rings include, but are not limited to, 1,2,3,4-tetrahydroisoquinolinyl, 1-oxo-1,2,3,4-tetrahydroisoquinolinyl, 1-oxo-1,2-dihydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridinyl, indolinyl, and isoindolinyl, wherein the heterocyclyl group may be attached through any ring of the fused system.

[0119] "Hydroxy" or "hydroxyl" refers to the group -OH. "Hydroxyalkyl" refers to an unbranched or branched alkyl group as defined above in which one or more hydrogen atoms are replaced by a hydroxy group.

[0120] "Oxo" refers to the group (=O) or (O).

[0121] "Nitro" refers to the radical -NO2.

[0122] "Sulfonyl" refers to the group -S(O)2R, where R is alkyl, haloalkyl, heterocyclyl, cycloalkyl, heteroaryl, or aryl. Examples of sulfonyl are methylsulfonyl, ethylsulfonyl, phenylsulfonyl, and toluenesulfonyl. "Alkylsulfonyl" refers to the group -S(O)2R, where R is alkyl.

[0123] "Alkylsulfinyl" refers to the group -S(O)R, where R is alkyl.

[0124] "Thiocyanate" refers to -SCN.

[0125] "Mercapto" refers to the group -SR, where R is alkyl, haloalkyl, heterocyclyl, cycloalkyl, heteroaryl, or aryl.

[0126] "Thio" or "thioketo" refers to the group (=S) or (S).

[0127] Some commonly used alternative chemical names may be used. For example, divalent groups such as divalent "alkyl" and divalent "aryl" may also be referred to as "alkylene" or "alkylenyl" or "arylene" or "arylenyl," respectively. In addition, unless otherwise defined, when a combination of groups is referred to herein as a moiety (e.g., arylalkyl), the last mentioned group contains the atom that connects the moiety to the rest of the molecule.

[0128] The term "1 to n" may refer to any positive integer from 1 to n, such as 1, 2, 3, 4, ..., n.

[0129] The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where said event or circumstance does not occur. Additionally, the term "optionally substituted" means that any one or more hydrogen atoms on a designated atom or group may or may not be replaced with a moiety other than hydrogen.

[0130] The term "substituted" means that any one or more hydrogen atoms on a designated atom or group are replaced by a moiety other than hydrogen, provided that the normal valence of the designated atom is not exceeded. The one or more substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amide, amidino, aryl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, guanidino, halogen, haloalkyl, haloalkoxy, heteroalkyl, heteroaryl, heterocyclyl, hydroxyl, hydrazine, imino, oxo, nitro, alkylsulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, sulfhydryl, thioketone, or a combination thereof. It is contemplated that polymers or similar indefinite structures obtained by further defining substituents with additional substituents are not included herein (e.g., substituted aryl with substituted alkyl, the substituted alkyl itself substituted with substituted aryl, the substituted aryl further substituted with substituted heteroalkyl, etc.). Unless otherwise stated, the maximum number of consecutive substitutions in the compounds described herein is 3. For example, the continuous substitution of a substituted aryl by two other substituted aryls is limited to an aryl substituted by a substituted aryl (substituted aryl). Similarly, the above definition is intended to exclude unallowed substitution patterns (e.g., methyl substituted with 5 fluorines or heteroaryl with two adjacent oxygen ring atoms). These unallowed substitution patterns are well known to those skilled in the art. When used to modify a chemical group, the term "substituted" may describe other chemical groups defined herein. Unless otherwise stated, when a group is described as optionally substituted, any substituent of the group is itself unsubstituted. For example, in some embodiments, the term "substituted alkyl" refers to an alkyl group with one or more substituents, including hydroxyl, halogen, alkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl. In other embodiments, the one or more substituents may be further substituted by halogen, alkyl, haloalkyl, hydroxyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, and each of the above groups is substituted. In other embodiments, the substituents may be further substituted with halogen, alkyl, haloalkyl, alkoxy, hydroxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is unsubstituted.

[0131] Active ingredient

[0132] The present invention provides a compound of formula I:

[0133] where X 1 、X 2 、X 3 、R 4 、R 5 and R 6 As described in this article.

[0134] Also provided are pharmaceutically acceptable salts, tautomers, stereoisomers, mixtures of stereoisomers, or isotopic substitutes thereof of the compounds described herein.

[0135] In many cases, the compounds of the present invention are capable of forming acid and / or base salts due to the presence of amino and / or carboxyl groups or similar groups. "Pharmaceutically acceptable salts" or "physiologically acceptable salts" include, for example, salts formed with inorganic acids or with organic acids. In addition, if the compounds described herein are obtained as acid addition salts, the free base can be obtained by alkalizing a solution of the acid salt. Conversely, if the product is a free base, an addition salt, specifically a pharmaceutically acceptable addition salt, can be produced according to conventional procedures for preparing acid addition salts from base compounds by dissolving the free base in a suitable organic solvent and treating the solution with an acid. Those skilled in the art will be aware of various synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Inorganic acids from which salts can be derived include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. By way of example only, salts derived from inorganic bases include sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, such as alkylamines (i.e., NH2(alkyl)), dialkylamines (i.e., HN(alkyl)2), trialkylamines (i.e., N(alkyl)3), substituted alkylamines (i.e., NH2(substituted alkyl)), di(substituted alkyl)amines (i.e., HN(substituted alkyl)2), tri(substituted alkyl)amines (i.e., N(substituted alkyl)3), alkenylamines (i.e., NH2(alkenyl)), dienylamines (i.e., HN(alkenyl)2), trialnylamines (i.e., N(alkenyl)3), substituted alkenylamines (i.e., NH2(substituted alkenyl)), di(substituted

[0014] The present invention also provides a substituted alkenyl)amine (i.e., HN(substituted alkenyl)2), a tri(substituted alkenyl)amine (i.e., N(substituted alkenyl)3), a mono-, di- or tri-cycloalkylamine (i.e., NH2(cycloalkyl), HN(cycloalkyl)2, N(cycloalkyl)3), a mono-, di- or tri-arylamine (i.e., NH2(aryl), HN(aryl)2, N(aryl)3), or a mixed amine, etc. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(isopropyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.

[0136] Some compounds exist in tautomer forms. Tautomers are in equilibrium with each other. For example, compounds containing amides can exist in equilibrium with imidic acid tautomers. No matter which tautomer is shown, and no matter how the nature of the equilibrium between the tautomers is, those of ordinary skill in the art will understand that the compound comprises amides and imidic acid tautomers. Therefore, compounds containing amides are understood to include their imidic acid tautomers. Similarly, compounds containing imidic acid are understood to include their amide tautomers.

[0137] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers, and individual isomers (e.g., isolated enantiomers) are intended to be encompassed within the scope of the present invention. When compounds provided herein have defined stereochemistry (denoted as R or S, or indicated by dashed or wedge-shaped bonds), those skilled in the art will understand that those compounds are substantially free of other isomers (e.g., at least 80%, 90%, 95%, 98%, 99%, and up to 100% free of other isomers).

[0138] Any formula or structure given in this application is also intended to represent unlabeled forms of the compound as well as isotopically labeled forms. An isotopically labeled compound has a structure described by the formula given in this application except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as, but not limited to, 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 15 N. 18 F. 31 P. 32 P. 35 S. 36 Cl and 125 I. The present invention includes various isotopically labeled compounds of the present invention, for example, those labeled with radioactive isotopes such as 3 H. 13 C and 14 Such isotopically labeled compounds are useful in metabolism studies, reaction kinetic studies, detection or imaging techniques, such as nuclear magnetic resonance imaging (NMR), positron emission tomography (PET), or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays, or in radiotherapy of patients.

[0139] The present disclosure also includes "deuterated analogs" of compounds of Formula I in which 1 to n (where n is the number of hydrogens in the molecule) hydrogens attached to carbon atoms are replaced by deuterium. Such compounds exhibit enhanced resistance to metabolism and are therefore useful for increasing the half-life of any compound of Formula I when administered to a mammal, particularly a human. Such compounds are synthesized by methods known in the art, for example, by using starting materials in which one or more hydrogen atoms have been replaced by deuterium.

[0140] Deuterium-labeled or substituted therapeutic compounds of the present invention can have improved DMPK (drug metabolism and pharmacokinetics) properties, which relate to distribution, metabolism and excretion (ADME). Using heavier isotope (e.g., deuterium) substitution can provide some therapeutic advantages based on larger metabolic stability, such as increased half-life in vivo, reduced dosage requirements and / or improved therapeutic index. 18 F-labeled compounds can be used for PET or SPECT studies. Isotopically labeled compounds of the present invention and their prodrugs are generally prepared by utilizing the synthetic routes described below or the steps described in the examples and preparations, wherein non-isotopically labeled reagents are replaced with readily available isotopically labeled reagents. It is understood that deuterium in this application can be considered a substituent of a compound of Formula I.

[0141] The concentration of such heavier isotopes (specifically deuterium) can be defined by an isotopic enrichment factor. In the compounds of the present invention, any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise indicated, when a position is specifically designated as "H" or "hydrogen," it is understood that the position is a hydrogen with its natural abundance isotopic composition. Thus, in the compounds of the present invention, any atom specifically designated as deuterium (D) is meant to represent deuterium.

[0142] The present invention also provides the compound or its pharmaceutically acceptable hydrates, solvates, polymorphs, active metabolites and prodrugs.

[0143] A "solvate" is formed by the interaction of a solvent and a compound. Also provided are solvates of the salts of the compounds described herein. Also provided are hydrates of the compounds described herein.

[0144] In certain embodiments, prodrugs of the compounds described herein are provided. "Prodrug" refers to any compound that, when administered to a biological system, produces a drug substance or active ingredient due to a spontaneous chemical reaction, an enzyme-catalyzed chemical reaction, photolysis, and / or metabolic chemical reaction. A prodrug is thus a covalently modified analog or latent form of a therapeutically active compound. Non-limiting examples of prodrugs include ester moieties, quaternary ammonium moieties, ethylene glycol moieties, and the like.

[0145] Also provided herein are in vivo metabolic products of the compounds described herein. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, esterification, etc. of the administered compound, primarily due to enzymatic processes.

[0146] use

[0147] Experiments have shown that the compounds of the present invention can specifically inhibit Osaka thyroid carcinoma (Cot) kinase, and thus can be used to prevent or treat diseases or symptoms mediated by Osaka thyroid carcinoma (Cot) kinase.

[0148] As used herein, "treat" refers to an agent used to obtain a beneficial or desired result (including a clinical result). Beneficial or desired clinical results may include one or more of the following: a) inhibiting a disease or condition (e.g., reducing one or more symptoms caused by the disease or condition and / or alleviating the severity of the disease or condition); b) slowing or preventing the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and / or preventing or delaying the spread of the disease or condition (e.g., metastasis)); and / or c) alleviating the disease, i.e., causing regression of clinical symptoms (e.g., improving the disease state, providing partial or complete relief of the disease or condition, enhancing the effects of other drug therapies, delaying disease progression, improving quality of life, and / or prolonging survival).

[0149] "Prevention" or "prevention" refers to any treatment of a disease or condition so that clinical symptoms of the disease or condition do not appear. In some embodiments, the compound can be administered to a subject (including a human) at risk for a disease or condition or with a family history of the disease or condition.

[0150] The term "therapeutically effective amount" or "effective amount" of a compound described herein, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof, refers to an amount sufficient to achieve treatment when administered to a subject, to provide a therapeutic benefit, such as improving symptoms or slowing disease progression. For example, a therapeutically effective amount may be an amount sufficient to reduce the symptoms of a disease or condition responsive to inhibition of Cot activity. The therapeutically effective amount may vary depending on the subject and the disease or condition being treated, the subject's weight and age, the severity of the disease or condition, and the mode of administration, which can be readily determined by one of ordinary skill in the art.

[0151] The term "inhibit" refers to a decrease in the baseline activity of a biological activity or process. "Inhibition of the activity of COT" or variations thereof refers to a decrease in the activity of COT in the presence of a compound of the invention, directly or indirectly, relative to the activity of COT in the absence of a compound of the invention. "Inhibition of COT" refers to a decrease in the activity of COT in the presence of a compound described herein, directly or indirectly, relative to the activity of COT in the absence of a compound described herein. In some embodiments, inhibition of COT activity can be compared to the same subject before treatment, or compared to other subjects not receiving the treatment.

[0152] The methods described herein can be applied to in vivo or ex vivo cell colonies. "In vivo" means within a living individual, such as an animal or human body. In this case, the methods described herein can be used therapeutically in an individual. "Ex vivo" means outside a living individual. Examples of ex vivo cell colonies include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from an individual. Such samples can be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. Exemplary tissue samples include tumors and biopsies thereof. In this context, the compounds and compositions described herein can be used for various purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used ex vivo to determine the optimal dosing schedule and / or dosage of Cot selective inhibitors for specified indications, cell types, individuals, and other parameters. The information collected from such uses can be used for experimental purposes or in the clinic to set an in vivo treatment regimen. Other ex vivo uses to which the compounds and compositions described herein may be suitable are described below or should be clear to those skilled in the art. The selected compounds can be further characterized to check safety or tolerable doses in humans or non-human subjects. Such properties can be examined using methods generally known to those skilled in the art.

[0153] The compounds disclosed herein can be used to prepare a medicament for preventing or treating a disease or condition mediated by Cot kinase, or for a method of treating a disease or condition mediated by Cot. Non-limiting examples of diseases or conditions mediated by Cot include, but are not limited to, cancer, diabetes, and inflammatory diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), inflammatory bowel disease (IBD), sepsis, psoriasis, dysregulated TNF expression, and transplant rejection.

[0154] In some embodiments, the disease or condition mediated by Cot is a solid tumor. In a specific embodiment, the solid tumor is selected from pancreatic cancer, bladder cancer, colorectal cancer, breast cancer, prostate cancer, kidney cancer, hepatocellular carcinoma, lung cancer, ovarian cancer, cervical cancer, stomach cancer, esophageal cancer, head and neck cancer, melanoma, neuroendocrine cancer, CNS cancer, brain tumor (e.g., glioma, anaplastic oligodendroglioma, adult glioblastoma multiforme, and adult anaplastic astrocytoma), bone cancer, or soft tissue sarcoma. In some embodiments, the solid tumor is selected from non-small cell lung cancer, small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, kidney cancer, prostate cancer, or breast cancer.

[0155] In some embodiments, the disease or condition mediated by Cot is diabetes, which includes any metabolic disorder characterized by impaired insulin production and glucose tolerance. In some embodiments, diabetes includes type 1 and type 2 diabetes, gestational diabetes, prediabetes, insulin resistance, metabolic syndrome, impaired fasting glucose, and glucose intolerance. Type 1 diabetes is also known as insulin-dependent diabetes mellitus (IDDM). Type 2 diabetes is also known as non-insulin-dependent diabetes mellitus (NIDDM).

[0156] In some embodiments, the disease or illness mediated by Cot is an inflammatory disease or LPS-induced endotoxic shock. In some embodiments, the disease is an autoimmune disease. In a specific embodiment, the autoimmune disease is systemic lupus erythematosus (SLE), myasthenia gravis, rheumatoid arthritis (RA), acute disseminated encephalomyelitis, idiopathic thrombocytopenic purpura, multiple sclerosis (MS), inflammatory bowel disease (IBD), sepsis, psoriasis, Sjögren's syndrome, autoimmune hemolytic anemia, asthma or chronic obstructive pulmonary disease (COPD), ankylosing spondylitis, acute gout and ankylosing spondylitis, reactive arthritis, monoarticular arthritis, osteoarthritis, gouty arthritis, juvenile arthritis, juvenile onset rheumatoid arthritis, juvenile rheumatoid arthritis or psoriatic arthritis. In other embodiments, the disease is inflammation. In other embodiments, the disease is an excessive or destructive immune response, such as asthma, rheumatoid arthritis, multiple sclerosis, chronic obstructive pulmonary disease (COPD), and lupus.

[0157] In some embodiments, the disease or condition mediated by Cot is inflammatory bowel disease (IBD).As used herein, the term "inflammatory bowel disease" or "IBD" is a general term describing inflammatory conditions of the gastrointestinal tract, the most common forms of which are ulcerative colitis and Crohn's disease. Other forms of IBD that can be treated with the compounds, compositions, and methods disclosed herein include diversion colitis, ischemic colitis, infectious colitis, chemical colitis, microscopic colitis (including collagenous colitis and lymphocytic colitis), atypical colitis, pseudomembranous colitis, fulminant colitis, autistic enterocolitis (autistic enterocolitis), indeterminate colitis, Behçet's disease, gastroduodenal CD, jejunoileitis, ileitis, ileocolitis, Crohn's (granulomatous) colitis, irritable bowel syndrome, mucositis, radiation-induced enteritis, short bowel syndrome, celiac disease, gastric ulcer, diverticulitis, pouchitis, proctitis, and chronic diarrhea.

[0158] Treatment or prevention of IBD also includes improving or alleviating one or more symptoms of IBD. As used herein, the term "symptoms of IBD" refers to detected symptoms, such as abdominal pain, diarrhea, rectal bleeding, weight loss, fever, loss of appetite and other more serious complications, such as dehydration, anemia and malnutrition. A variety of these symptoms are subjected to quantitative analysis (e.g., weight loss, fever, anemia, etc.). Some symptoms are easily determined based on blood tests (e.g., anemia) or tests that detect the presence of blood (e.g., rectal bleeding). The term "wherein alleviating these symptoms" refers to a qualitative or quantitative alleviation of detectable symptoms, including, but not limited to, a detectable effect on the rate of recovery from the disease (e.g., rate of weight gain). Diagnosis is typically determined by endoscopic observation of the mucosa and pathological examination of endoscopic biopsy samples.

[0159] The course of IBD varies and is generally associated with intermittent periods of remission and exacerbation. Various methods have been described to characterize the disease activity and severity of IBD and the response to treatment of subjects with IBD. Treatment according to the methods of the present invention is generally applicable to subjects with IBD at any level or degree of disease activity.

[0160] In some embodiments, the diseases or conditions treated by administering a compound of the compositions described herein include acute gout and ankylosing spondylitis, allergic diseases, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis and multiple sclerosis, atherosclerosis, bacterial infections, bone cancer pain and pain caused by endometriosis, BRAF-resistant melanoma, brain stem glioma or pituitary adenoma, burns, bursitis, cancer of the anal region, cancer of the endocrine system, cancer of the kidney or ureter (e.g., renal cell carcinoma, renal pelvis cancer), cancer of the penis, cancer of the small intestine, cancer of the thyroid gland, cancer of the urethra, blood cancers such as acute myeloid leukemia, cancer of the tongue, cancer of the cervix, cancer of the endometrium, cancer of the fallopian tube, cancer of the renal pelvis, cancer of the vagina or vulva, chronic myeloid leukemia, chronic or acute leukemia, chronic pain, classic Bartter syndrome (e.g., Bartter syndrome), syndrome), common cold, conjunctivitis, coronary heart disease, skin or intraocular melanoma, dermatitis, dysmenorrhea, eczema, endometriosis, familial adenomatous polyposis, fibromyalgia, fungal infection, gout, gynecological cancer, uterine sarcoma, fallopian tube cancer, headache, hemophilic arthropathy, Parkinson's disease, AIDS, herpes zoster, Hodgkin's disease, Huntington's disease, hyperprostaglandin E syndrome, influenza, iritis, juvenile arthritis, juvenile-onset rheumatoid arthritis, juvenile rheumatoid arthritis, low back and neck pain, lymphocytic lymphoma, myofascial disorders, Myositis, neuralgia, neurodegenerative disorders such as Alzheimer's disease, neuroinflammatory disorders, neuropathic pain, vulvar cancer, Parkinson's disease, pediatric malignancies, pulmonary fibrosis, rectal cancer, rhinitis, sarcoidosis, soft tissue sarcoma, scleritis, skin cancer, solid tumors in children, spinal cord tumors, sprains and strains, gastric cancer, stroke, subacute and chronic musculoskeletal pain syndromes such as bursitis, surgical or dental procedures, symptoms associated with influenza or other viral infections, synovitis, toothache, ulcers, uterine cancer, uterine sarcoma, uveitis, vasculitis, viral infections, viral infections (e.g., influenza), and wound healing.

[0161] In some embodiments, the Cot-mediated disease or condition is alcoholic hepatitis. Alcoholic hepatitis is a clinical syndrome characterized by jaundice and liver failure in subjects with chronic and active alcohol abuse (see Akriviadis E. et al., Ann Gastroenterol. 2016 Apr-Jun; 29(2): 236-237). Alcoholic hepatitis can cause cirrhosis and hepatocellular fibrosis. Glucocorticoids (e.g., prednisolone) and phosphodiesterase inhibitors (e.g., pentoxifylline) can be used to treat alcoholic hepatitis. The compounds herein can be used as a single treatment or in combination with current alcoholic hepatitis treatments.

[0162] In some embodiments, the Cot-mediated disease or condition is systemic lupus erythematosus (SLE), lupus nephritis, lupus-related or other autoimmune diseases or SLE symptoms. Symptoms of systemic lupus erythematosus include joint pain, joint swelling, arthritis, fatigue, hair loss, mouth ulcers, swollen lymph nodes, sensitivity to sunlight, rash, headache, numbness, tingling, seizures, vision problems, personality changes, abdominal pain, nausea, vomiting, abnormal heart rhythm, coughing up blood and difficulty breathing, patchy skin color and Raynaud's phenomenon.

[0163] "Subject" refers to an animal, such as a mammal (including a human), that has been or will be the subject of treatment, observation, or experiment. The methods described herein can be used for human therapy and veterinary applications. In some embodiments, the subject is a mammal. In one embodiment, the subject is a human.

[0164] Combination therapy

[0165] In one embodiment, the compounds disclosed herein can be used in combination with one or more other therapeutic agents being used and / or developed to treat inflammatory disorders (e.g., IBD). The one or more additional therapeutic agents can be an α4β7 inhibitor, a steroid, an MMP-9 antibody, an S1P1 agonist, a TNF biologic, or any combination thereof.

[0166] In some embodiments, the one or more other therapeutic agents may be an α4β7 integrin inhibitor, or an agent that inhibits the expression and / or activity of the α4β7 integrin. The inhibitor may be a small molecule or a biologic. For example, the α4β7 integrin inhibitor may be natalizumab or vedolizumab.

[0167] In some embodiments, the one or more additional therapeutic agents may be steroids, including but not limited to corticosteroids. Corticosteroids can be administered by a variety of routes, including intravenous (i.e., methylprednisolone, hydrocortisone), oral (i.e., prednisone, prednisolone, budesonide, dexamethasone), or topical (i.e., enema, suppository, or foam formulation).

[0168] In some embodiments, the one or more other therapeutic agents may be a TNF inhibitor, or an agent that inhibits the expression and / or activity of TNF. The inhibitor may be a small molecule or a biologic. For example, the TNF inhibitor may be golimumab.

[0169] In some embodiments, the one or more additional therapeutic agents are being used and / or developed to treat ulcerative colitis (UC) and / or Crohn's disease (CD).

[0170] In some embodiments, the one or more additional therapeutic agents may be a JAK inhibitor, particularly a JAK-1 selective inhibitor. The inhibitor may be a small molecule or a biologic. For example, the JAK inhibitor may be filgotinib or GLPG0634 (a JAK modulator from Galápagos).

[0171] Pharmaceutical composition and administration method

[0172] The compound provided herein is usually administered in the form of a pharmaceutical composition. Therefore, a pharmaceutical composition is also provided herein, which comprises one or more compounds described herein or their pharmaceutically acceptable salts, tautomers, stereoisomers, stereoisomer mixtures, prodrugs or their isotope substitutes, and one or more pharmaceutically acceptable vehicles selected from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable carriers may include, for example, inert solid diluents and fillers, diluents (including sterile aqueous solutions and various organic solvents), penetration enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner known to the pharmaceutical field.

[0173] "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials that are suitable for use in preparing pharmaceutical compositions for veterinary or human medical use.

[0174] As used herein, "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings / coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. Such media and agents for pharmaceutically active substances are well known in the art. Unless any conventional media or agents are incompatible with the active ingredient, their use in therapeutic compositions is contemplated. Supplementary active ingredients may also be incorporated into the compositions.

[0175] The pharmaceutical composition can be administered in a single dose or multiple doses. The pharmaceutical composition can be administered by various methods, including, for example, rectal, buccal, intranasal, and transdermal routes. In certain embodiments, the pharmaceutical composition can be administered by intraarterial injection, intravenous, intraperitoneal, parenteral, intramuscular, subcutaneous, oral, topical, or inhaled form.

[0176] One mode of administration is parenteral, for example, by injection. Forms that can be incorporated into the pharmaceutical compositions described herein for administration by injection include, for example, aqueous or oily suspensions or emulsions containing sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or sterile aqueous solutions and similar pharmaceutical vehicles.

[0177] Oral administration can be another route for administering the compounds described herein. For example, administration can be via capsules or enteric-coated tablets. In preparing pharmaceutical compositions comprising at least one compound described herein, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof, the active ingredient is typically diluted with an excipient and / or encapsulated in a carrier that can be in the form of a capsule, pouch, paper, or other container. When an excipient is used as a diluent, it can be in the form of a solid, semisolid, or liquid material (as described above) that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the composition can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (in solid form or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.

[0178] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, sterile water, syrup and methylcellulose. The formulation may additionally include: lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners; and flavoring agents.

[0179] The composition comprising at least one compound as described herein or its pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue can be formulated so as to provide rapid, sustained or delayed release of the active ingredient after administration to the subject by adopting operations known in the art. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolution systems, which contain polymer-coated reservoirs or drug-polymer matrix formulations. Another formulation used in the method disclosed herein adopts a transdermal delivery device ("patch"). Such transdermal patches can be used to provide compounds as described herein for continuous or discontinuous infusion in controlled amounts.

[0180] For the preparation of solid compositions such as tablets, the principal active ingredient can be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analog thereof. When referring to these preformulation compositions as homogeneous compositions, the active ingredient is dispersed evenly throughout the composition so that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

[0181] The tablets or pills of the compounds described herein can be coated or otherwise mixed to provide a dosage form that has an advantage of prolonged action time or protection from the acidic conditions of the stomach. For example, a tablet or pill can include an inner dose and an outer dose component, the latter being an encapsulated form on the former. The two components can be separated by an enteric layer to resist disintegration in the stomach and to allow the inner component to enter the duodenum intact or to delay release. Many materials can be used as the enteric layer or coating, including many polymeric acids, and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0182] Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein. In some embodiments, the compositions are administered by oral or nasal respiratory routes for local or systemic effect. In other embodiments, the compositions in pharmaceutically acceptable solvents may be aerosolized using an inert gas. The aerosolized solution may be inhaled directly from the aerosolizing device or the aerosolizing device may be connected to a mask holder or an intermittent positive pressure breathing machine. The solution, suspension, or powder composition may preferably be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0183] Drug administration

[0184] For any specific subject, the specific dosage level of the compound of the present application will depend on a variety of factors, including the activity of the specific compound used in the subject undergoing therapy, age, body weight, overall health, sex, diet, administration time, route of administration and excretion rate, drug combination and the severity of the specific disease. For example, the dosage can be expressed as milligrams of the compound described herein per kilogram of subject body weight (mg / kg). A dosage of about 0.1 to 150 mg / kg may be appropriate. In some embodiments, about 0.1 to 100 mg / kg may be appropriate. In other embodiments, a dosage of 0.5 to 60 mg / kg may be appropriate. When adjusting the dosage between subjects with widely different body sizes, for example, when using the drug in children and adults or when converting the effective dose in non-human subjects (e.g., dogs) to a dosage suitable for human subjects, standardization according to the subject's weight is particularly applicable.

[0185] The daily dose can also be described as the total amount of a compound described herein administered per dose or per day. The daily dose of a compound of Formula I can be about 1 mg to 4,000 mg, about 2,000 to 4,000 mg / day, about 1 to 2,000 mg / day, about 1 to 1,000 mg / day, about 10 to 500 mg / day, about 20 to 500 mg / day, about 50 to 300 mg / day, about 75 to 200 mg / day, or about 15 to 150 mg / day.

[0186] When administered orally, the total daily dose for a human subject can be 1 mg to 1,000 mg, about 1,000-2,000 mg / day, about 10-500 mg / day, about 50-300 mg / day, about 75-200 mg / day, or about 100-150 mg / day.

[0187] The compounds of the present application or compositions thereof can be administered once, twice, three times or four times daily using any suitable mode described above. In addition, the administration or treatment of the compound can be continuous for multiple days; for example, for a treatment cycle, the treatment will generally last for at least 7 days, 14 days or 28 days. Treatment cycles are well known in cancer chemotherapy and are often alternating with rest periods of about 1 to 28 days, typically about 7 days or about 14 days, between multiple cycles. In other embodiments, the treatment cycles can also be continuous.

[0188] In one embodiment, the method comprises administering to a subject an initial daily dose of about 1 to 800 mg of a compound described herein and increasing the dose in increments until clinical efficacy is achieved. The dose may be increased in increments of about 5, 10, 25, 50, or 100 mg. The dose may be increased daily, every other day, twice a week, or once a week.

[0189] Preparation method, intermediates

[0190] The compounds of the present invention have no particular requirements for the preparation method and can be prepared according to their chemical structures by referring to commonly used organic synthesis methods in the art, or by referring to the methods described below in the present invention.

[0191] The present invention also provides a key intermediate that can be used to prepare the compound of the present invention:

[0192] Among them, R 16 is Cl, Br, I, OTf or NH2;

[0193] Among them, X 1 、X 2 、X 3 、R 1 、R 2 、R 5 and R 16As defined herein.

[0194] The main advantages of the present invention include:

[0195] 1. The present invention provides a novel class of Osaka thyroid carcinoma (Cot) kinase inhibitors.

[0196] 2. The compounds of the present invention have excellent Osaka thyroid carcinoma (Cot) kinase inhibitory activity and can specifically inhibit Osaka thyroid carcinoma (Cot) kinase, thereby preventing or / treating related diseases.

[0197] 3. The compounds of the present invention have good drugability and are very suitable for preparing drugs for preventing or / treating diseases or conditions mediated by Osaka thyroid carcinoma (Cot) kinase.

[0198] The present invention will be further described below in conjunction with specific implementation. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0199] Synthesis process

[0200] Abbreviation Definition

[0201] Certain abbreviations are defined as follows: “°C” means degrees Celsius; “Ac” means acetyl; “ATP” means adenosine triphosphate; “Boc” means tert-butyloxycarbonyl; “br” means broad; “BSA” means bovine serum albumin; “Cbz” means carbonylbenzyl; “ChEF” means chelate-enhanced fluorescence; “COPD” means chronic obstructive pulmonary disease; “Cot” means Osaka thyroid carcinoma; “d” means doublet; “DCM” means dichloromethane; “dd” means doublet of doublet; “DIPEA” means N,N-diisopropylethylamine; “DMF” means N,N-dimethylformamide; “DMSO” means dimethyl sulfoxide; “dppf” means 1,1′-bis(diphenylphosphino)ferrocene; “dt” means doublet of triplet; “DTT” means dithiothreitol; “EA” means ethyl acetate; “EGF” means iodine. “R” refers to epidermal growth factor receptor; “EGTA” refers to ethylene glycol bis(2-aminoethyl ether) tetraacetic acid; “eq” refers to equivalents; “ES / MS” refers to electrospray ionization mass spectrometry; “Et” refers to ethyl; “FA” refers to formic acid; “g” refers to grams; “HATU” refers to 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; “HEPES” refers to 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; “HPLC” refers to high pressure liquid chromatography; “Hz” refers to hertz; “IBD” refers to inflammatory bowel disease; “i-Pr” refers to isopropyl; “J” refers to coupling constant (MHz); “LPS” refers to lipopolysaccharide; “LTBA” refers to lithium tri-tert-butyloxyaluminum hydride; “MS” refers to mass spectrometry; “M” refers to molarity; “m” refers to multiplet; “M+H” refers to chromatographic markers. + ” refers to proton peak + hydrogen; “Me” refers to methyl; “mg” refers to milligram; “MHz” refers to megahertz; “min” refers to minute; “mL” refers to milliliter; “mM” refers to millimolar; “mmol” refers to millimolar; “MS” refers to mass spectrometry; “Ms” refers to methanesulfonyl; “nBu / Bu” refers to butyl; “NBS” refers to N-bromosuccinimide; “nL” refers to nanoliter; “nm” refers to nanometer; “NMR” refers to nuclear magnetic resonance; “Pd / C” refers to palladium / carbon; “PE” refers to petroleum ether; “ "Ph" refers to phenyl; "PS" refers to polystyrene; "q" refers to quartet; "rt" refers to room temperature; "s" refers to singlet; "t" refers to triplet; "t-Bu" refers to tert-butyl; "Tf" refers to trifluoromethanesulfonyl; "TFA" refers to trifluoroacetic acid; "TFAA" refers to trifluoroacetic anhydride; "THF" refers to tetrahydrofuran; "Tpl2" refers to tumor progression locus 2; "TNF" refers to tumor necrosis factor; "δ" refers to chemical shift (ppm); "μL" refers to microliter; "μM" refers to micromolar;

[0202] Process 1

[0203] In Scheme 1, Step A shows that compound 2 can be provided by reacting an appropriately substituted aldehyde 1 with an acetylene Grignard reagent, followed by acylation of the resulting alcohol with acetic anhydride.

[0204] Process 2

[0205] In Scheme 2, Step A describes the treatment of an appropriately substituted amine 3 with a diazo transfer reagent (e.g., imidazole-1-sulfonyl azide hydrochloride) to yield the corresponding compound 4. Step B shows that an appropriately substituted triazole 5 can be prepared by reacting compound 4 with methyl propiolate via standard 1,3-dipolar cycloaddition conditions. Step C describes the hydrolysis of compound 5 in a suitable inorganic base (e.g., NaOH, etc.) in a suitable solvent (e.g., THF, H2O, etc.) to yield compound 6. Step D describes the reaction of compound 6 with an inorganic base (e.g., sodium acetate, etc.) in a solvent (e.g., H2O, etc.) at an elevated temperature (e.g., about 75°C) in the presence of a brominating agent (e.g., NBS, etc.) to yield compound 7. Step E shows the preparation of compound 8 by the following method: compound 7 is reacted with compound 1 in a solvent (e.g., THF, etc.) in the presence of an organometallic reagent (e.g., isopropylmagnesium chloride lithium chloride complex, etc.) to yield compound 8. Step F shows that compound 8 is treated with an oxidizing agent (e.g., Dess-Martin reagent, etc.) to obtain compound 9. Step G describes the reaction of compound 9 with tert-butylsulfenamide in the presence of a titanium- or zirconium-based reagent in a suitable solvent (e.g., toluene, etc.) at a suitable temperature (e.g., about 75°C) to obtain compound 10. Step H describes the treatment of compound 10 with a reducing agent (e.g., LTBA, etc.) to obtain compound 11. Step I shows that compound 11 is reacted with an acid (e.g., concentrated hydrochloric acid, etc.) in a suitable solvent (e.g., methanol, etc.) at a suitable temperature (e.g., about 50°C) to obtain compound 12.

[0206] Process 3

[0207] In Scheme 3, Step A describes the reaction of compound 13 in the presence of a halogenating agent (e.g., NCS, etc.) in a suitable solvent (e.g., DMF, etc.) to produce compound 14. Step B shows the solvolysis reaction of compound 14 in a suitable ammonia solution (e.g., ammonia-methanol, etc.) to produce compound 15. Step C describes the condensation reaction of compound 15 with DMF·DMA at a suitable temperature (e.g., 100°C) and a suitable solvent (e.g., 1,4-dioxane, etc.) to produce compound 16. Step D describes the intramolecular cyclization reaction of compound 16 at a suitable temperature (e.g., 100°C) in the presence of a suitable base (e.g., cesium carbonate, etc.) and a suitable solvent (e.g., 1,4-dioxane, etc.) to produce compound 17. Step E shows the condensation reaction of compound E with an amine in a suitable condensing agent (e.g., PyBOP, etc.), a suitable base (e.g., DIPEA, etc.), and a suitable solvent (e.g., DMF, etc.) to produce compound 18. In step F, Buchwald coupling can be carried out in a suitable solvent (e.g., 1,4-dioxane) at a suitable temperature (e.g., about 120° C.) using a suitable catalyst (e.g., Xantphos Pd G2, etc.) and a suitable base (e.g., sodium tert-butoxide, etc.) to react compound 18 with compound 12 to obtain compound 19.

[0208] Intermediate preparation

[0209] Intermediate 1

[0210] Preparation of 1-(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)prop-2-en-1-yl acetate (I001)

[0211] Step 1:

[0212] 5-Formyl-2-methylisoquinolin-2-ium (I001b)

[0213] At room temperature, I001a (20 g, 127 mmol) was dissolved in MeCN (120 mL), and CH3I (36 g, 254 mmol) was slowly added. The mixture was stirred at room temperature for 12 hours. After completion of the reaction, the mixture was filtered, and the filter cake was washed with toluene (30 mL x 3) and dried at approximately 30°C to obtain compound I001b (34 g, 90% yield). The crude product was used directly in the next reaction without further purification. MS m / z (ESI): 172.2 [M+H] + .

[0214] Step 2:

[0215] 2-Methyl-1-oxo-1,2-dihydroisoquinoline-5-carbaldehyde (I001c)

[0216] I001b (34 g, 114 mmol) and potassium ferricyanide (75 g, 228 mmol) were dissolved in H₂O (300 mL). Under nitrogen, a KOH (25.5 g, 456 mmol) aqueous solution (200 mL) was slowly added to the reaction solution, followed by reaction at 5°C for 2 hours. After completion of the reaction, the mixture was filtered, and the resulting filter cake was washed with H₂O (100 mL x 3). The crude product was purified by silica gel column chromatography (PE:EA = 1:1) to afford compound I001c (13 g, 61.9% yield). MS m / z (ESI): 188.0 [M+H] + .

[0217] Step 3:

[0218] 1-(2-Methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)prop-2-en-1-yl acetate (I001)

[0219] I001c (4.0 g, 21.4 mmol) was dissolved in THF (120 mL). Ethynylmagnesium bromide (85.6 mL, 42.8 mmol) was slowly added with stirring under nitrogen displacement. The mixture was allowed to react at 0°C for 4 hours. Acetyl chloride (3.4 g, 42.8 mmol) was then added dropwise and allowed to react for 4 hours. After completion, the reaction was quenched with saturated aqueous sodium bicarbonate (60 mL) and extracted with EA (80 mL x 3). The combined organic phases were washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product, which was purified by silica gel column chromatography (PE:EA = 1:2) to afford compound I001 (4.3 g, 79.6% yield). MS m / z (ESI): 256.1 [M+H] + .

[0220] Intermediate 2

[0221] Preparation of (S)-5-{amino[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I002)

[0222] Step 1:

[0223] 1-Azidobicyclo[1.1.1]pentane (I002b)

[0224] I002a (5 g, 41.8 mmol) was dissolved in acetonitrile (60 mL), and DIPEA (10.8 g, 83.6 mmol) was added. ADMP (15.2 g, 54.4 mmol) was added in batches. The mixture was reacted at 20°C under nitrogen for 12 hours to obtain a solution of compound I002b, which was used directly in the next step.

[0225] Step 2:

[0226] Methyl [1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)acetate (I002c)

[0227] I001 (4 g, 15.6 mmol), copper sulfate pentahydrate (390 mg, 1.56 mmol), and sodium ascorbate (1.54 g, 7.8 mmol) were added to 2-methyltetrahydrofuran (100 mL). An acetonitrile solution of I002b was then added, and the mixture was allowed to react at 75°C under nitrogen for 24 hours. After completion, the reaction solution was concentrated, and DCM (100 mL) and water (50 mL) were added. The mixture was filtered, and the filtrate was separated. The aqueous phase was extracted twice with DCM. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by normal phase column chromatography (DCM:MeOH = 10:1) to afford compound I002c (3.2 g, 56.3% yield). MS m / z (ESI): 729.2 [2M+H] + .

[0228] Step 3:

[0229] 5-{[1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](hydroxy)methyl}-2-methylisoquinolin-1(2H)-one (I002d)

[0230] I002c (3.2 g, 8.79 mmol) was dissolved in THF (50 mL), and H₂O (10 mL) and NaOH (700 mg, 17.6 mmol) were added. The mixture was allowed to react at 20°C for 12 hours. After completion of the reaction, the pH was adjusted to 5-6 with 1.0 N₄Cl. The THF was concentrated to remove most of the aqueous phase, and the aqueous phase was extracted with DCM (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound I002d (2.4 g, 85.0% yield). MS m / z (ESI): 323.1 [M+H] + .

[0231] Step 4:

[0232] 5-[1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazole-4-carbonyl]-2-methylisoquinolin-1(2H)-one (I002e)

[0233] I002d (2.75 g, 8.56 mmol) was dissolved in anhydrous DCM (40 mL), and Dess-Martin reagent (5.44 g, 12.84 mmol) was added portionwise. The mixture was allowed to react at 30°C for 4 hours. After completion, the reaction was quenched with saturated aqueous sodium bicarbonate (50 mL) and extracted with DCM (50 mL x 2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (PE:EA = 2:1) to obtain compound I002e (2.60 g, 93% yield). 1 H NMR (400MHz, DMSO-d6) δ9.03 (s, 1H), 8.48 (d, J = 7.9Hz, 1H), 8.12 (d, J = 6.7Hz, 1H), 7.63 (t, J = 7. 8Hz,1H),7.54(d,J=7.7Hz,1H),6.77(d,J=7.6Hz,1H),3.53(s,3H),2.75(s,1H),2.40(s,6H).MS m / z(ESI):321.1[M+H] + .

[0234] Step 5:

[0235] (R,E)-N-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)methylene}-2-methylpropane-2-sulfamide (I002f)

[0236] I002e (2.6 g, 8.1 mmol) and (R)-(+)-tert-butylsulfenamide (2.94 g, 24.3 mmol) were dissolved in anhydrous toluene (50 mL) under nitrogen atmosphere. Tetraethyl titanate (5.54 g, 24.3 mmol) was added and reacted at 75°C for 16 hours. After completion of the reaction, the temperature was cooled to room temperature, concentrated, and purified by column chromatography (PE:EA = 1:1) to afford compound I002f (2.8 g, 81.7% yield). MS m / z (ESI): 424.1 [M+H] + .

[0237] Step 6:

[0238] (R)-N-{(S)-[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)methyl}-2-methylpropane-2-sulfonamide (1002g)

[0239] I002f (2.8 g, 6.61 mmol) was dissolved in anhydrous DCM (50 mL) under nitrogen protection, with the temperature maintained below 0°C. A solution of lithium tri-sec-butylborohydride in tetrahydrofuran (1.0 N, 13.2 mL, 13.2 mmol) was slowly added dropwise. The reaction was allowed to react at 0°C for 4 hours. After translation, saturated aqueous ammonium chloride was slowly added dropwise to quench the reaction. The aqueous phase was extracted twice with DCM. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound I002g (2.3 g, 82% yield). MS m / z (ESI): 426.1 [M+H] + .

[0240] Step 7:

[0241] (S)-5-{amino[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I002)

[0242] I002 g (2.3 g, 5.42 mmol) was dissolved in methanol (10 mL), and aqueous HCl (40 mL, 0.1 N) was added. The mixture was allowed to react at 50°C for 24 hours. After completion of the reaction, the temperature was lowered to 10°C, the pH was adjusted to 7, and the mixture was concentrated. The crude product was isolated and purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound I002 (1.05 g, 60.3% yield). 1 H NMR(400MHz,DMSO-d6)δ8.16 (d,J=7.8Hz,1H),7.89(d,J=6.4Hz,2H),7.52-7.41(m,2H),6.84(d,J=7.7Hz,1H),5.71(s,1H),3.49(s,3H),2.66(s,1H),2.28(s,6H).MS m / z(ESI):643.1[2M+H] + .

[0243] Intermediate 3

[0244] Preparation of 6-bromo-8-chloro-N-neopentylpyrido[3,4-d]pyrimidin-4-amine (I003)

[0245] Step 1:

[0246] 3-Amino-6-bromo-2-chloroisonicotinate (I003b)

[0247] Compound I003a (5 g, 21.6 mmol) was dissolved in anhydrous DMF (60 mL), and NCS (5.7 g, 43.3 mmol) was added. Stirring was continued at 20°C overnight. H2O (100 mL) was added to the reaction mixture for dilution, followed by extraction with EA (60 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product. Purification by silica gel column chromatography (PE:EA = 1:1) afforded compound I003b (5.2 g, 91.2% yield) as a yellow solid. MS m / z (ESI): 265 / 267 [M+H] + .

[0248] Step 2:

[0249] 3-Amino-6-bromo-2-chloroisonicotinamide (I003c)

[0250] Compound I003b (5.2 g, 19.6 mmol) was dissolved in ammonia-methanol (7.0 M, 100 mL) and the mixture was stirred overnight at 20°C in a sealed tube. The reaction was concentrated to afford compound I003c (4.5 g, 91.8% yield) as a yellow solid. MS m / z (ESI): 250 / 252 [M+H] + .

[0251] Step 3:

[0252] 3-Amino-6-bromo-2-chloro-N-[(dimethylamino)methylene]isonicotinamide (I003d)

[0253] Compound I003c (4.5 g, 18 mmol) was dissolved in anhydrous 1,4-dioxane (60 mL), and DMF-DMA (2.35 g, 19.8 mmol) was added. The reaction was stirred at 100°C under nitrogen for 1 hour. After completion of the reaction, the temperature was cooled to room temperature, the reaction solution was concentrated, washed with H2O (50 mL), and extracted with EA (100 mL). The organic phases were combined and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound I003d (3.5 g, 63.7% yield) as a yellow solid. MS m / z (ESI): 305 / 307 [M+H] + Step 4:

[0254] 6-Bromo-8-chloropyrido[3,4-d]pyrimidin-4-ol (I003e)

[0255] Compound I003d (2.5 g, 8.2 mmol) and cesium carbonate (8.0 g, 24.6 mmol) were dissolved in anhydrous 1,4-dioxane (100 mL) and stirred at 100°C under nitrogen for 4 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was poured into H2O (100 mL) and the pH was adjusted to 3-4 with 1.0 N dilute hydrochloric acid. A pale yellow solid precipitated, which was filtered and dried to obtain compound I003e (1.5 g, 71.4% yield). MS m / z (ESI): 260 / 262 [M+H] + .

[0256] Step 5:

[0257] 6-Bromo-8-chloro-N-neopentylpyrido[3,4-d]pyrimidin-4-amine (I003)

[0258] Compound I003e (1.4 g, 5.4 mmol), PyBOP (5.6 g, 10.8 mmol), and DIPEA (2.1 g, 16.2 mmol) were dissolved in DMF (50 mL). The system was cooled to approximately 0°C, and neopentylamine (1.8 g, 21.6 mmol) was added. The reaction was stirred overnight. H2O (50 mL) was added for dilution, and the mixture was extracted with EA (100 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was then purified by silica gel column chromatography (PE:EA = 1:1) to afford Compound I003 (700 mg, 39.4% yield) as a yellow solid. MS m / z (ESI): 329 / 331 [M+H] + .

[0259] Intermediate 4

[0260] Preparation of 1-(6-fluoro-2-methylpyridin-3-yl)prop-2-yn-1-yl acetate (I004)

[0261] Step 1:

[0262] (6-Fluoro-2-methylpyridin-3-yl)methanol (I004b)

[0263] Compound I004a (5 g, 32.25 mmol) was dissolved in THF (50 mL). BH3-Me2S (64.5 mL, 129.0 mmol) was added dropwise at 0°C. The mixture was allowed to react at 30°C for 16 h. After completion of the reaction, the temperature was lowered to 0°C, and MeOH (50 mL) was slowly added dropwise. Stirring was continued for 30 minutes. The mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound I004b (3.97 g, 88.2% yield). MS m / z (ESI): 142.1 [M+H] + .

[0264] Step 2:

[0265] 6-Fluoro-2-methylnicotinaldehyde (I004c)

[0266] Compound I004b (3.97 g, 28.12 mmol) was dissolved in THF (40 mL), and manganese dioxide (36.2 g, 422.3 mmol) was added portionwise. The mixture was stirred at 70°C under nitrogen for 16 hours. After completion of the reaction, the mixture was cooled to room temperature and filtered. The filter cake was washed with MeOH, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to obtain compound I004c (1.92 g, 49.1% yield). MS m / z (ESI): 140.1 [M+H] + .

[0267] Step 3:

[0268] 1-(6-Fluoro-2-methylpyridin-3-yl)prop-2-yn-1-yl acetate (I004)

[0269] Compound I004c (1.92 g, 13.8 mmol) was dissolved in THF (10 mL). Ethynylmagnesium bromide (55.2 mL, 27.6 mmol) was added dropwise at 0°C under nitrogen. The mixture was allowed to react at 0°C for 1 hour. Acetyl chloride (2.16 g, 27.6 mmol) was then added, and the reaction mixture was allowed to react at 30°C for 12 hours. After completion, the reaction mixture was poured into H2O (30 mL) and extracted with EA (3 x 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to obtain compound I004 (2.02 g, 70.8% yield). MS m / z (ESI): 208.1 [M+H] + .

[0270] Intermediate 5

[0271] Preparation of (S)-{1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methanamine (I005)

[0272] Step 1:

[0273] 1-Azido-1,1'-bicyclopropane (I005b)

[0274] Compound I005a (1.00 g, 7.46 mmol) was dissolved in acetonitrile (10 mL) under nitrogen atmosphere, with the reaction temperature maintained below 10°C. DIPEA (2.16 g, 16.4 mmol) was added dropwise to the reaction solution, stirred for 0.5 hour, and then 2-azido-1,3-dimethylimidazolium hexafluorophosphate (2.96 g, 10.4 mmol) was added. The reaction solution was slowly warmed to room temperature and stirred for 16 hours. After the reaction, the solution was used directly in the next step without further treatment to obtain an acetonitrile solution of compound I005b.

[0275] Step 2:

[0276] {1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methyl acetate (I005c)

[0277] Compound I005b (1.4 g, 6.76 mmol), 1-azido-1,1'-bicyclopropane (0.746 M, 10 mL), copper sulfate pentahydrate (845 mg, 3.38 mmol), and sodium ascorbate (669 mg, 3.38 mmol) were dissolved in 2-methyltetrahydrofuran (25 mL). After addition, the reaction mixture was stirred at 75°C under nitrogen overnight. After completion, the reaction was cooled to room temperature, water (20 mL) was added, and extraction with EA (20 mL x 3) was performed. The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (EA:PE = 2:1) to obtain compound I005c (1.5 g, 67.2% yield). MS m / z (ESI): 331.0 [M+H] + .

[0278] Step 3:

[0279] {1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methanol (I005d)

[0280] Compound I005c (1.50 g, 4.55 mmol) was dissolved in anhydrous THF (12 mL) at room temperature, and a solution of lithium hydroxide monohydrate (191 mg, 9.10 mmol) in water (2 mL) was added dropwise to the reaction mixture. The reaction was stirred at room temperature for 3 hours. After completion of the reaction, the pH of the reaction mixture was adjusted to 7-8 with 1.0 M hydrochloric acid solution. Water (10 mL) was added, and the mixture was extracted with EA (30 mL). The organic phases were combined, dried, and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain Compound I005d (1.20 g, 91.6% yield) as a yellow solid. MS m / z (ESI): 289.1 [M+H] + .

[0281] Step 4:

[0282] {1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methanone (I005e)

[0283] Compound I005d (1.20 g, 4.17 mmol) was dissolved in anhydrous DCM (20 mL), and Dess-Martin periodinane (3.54 g, 8.34 mmol) was added. The reaction system was stirred at room temperature overnight. After completion of the reaction, the reaction solution was filtered, and the filtrate was quenched with saturated aqueous sodium bicarbonate (10 mL) and extracted with DCM (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (PE:EA = 2:1) to obtain compound I005e (1.10 g, yield 92.2%). MS m / z (ESI): 287.2 [M+H] + .

[0284] Step 5:

[0285] (R,E)-N-{{1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methylene}-2-methylpropane-2-sulfenamide (I005f)

[0286] Compound I005e (1.10 g, 3.85 mmol) was dissolved in toluene (10 mL), and tetraethyl titanate (3.51 g, 15.4 mmol) and (R)-(+)-tert-butylsulfenamide (1.40 g, 11.6 mmol) were added. The reaction mixture was stirred at 75°C under nitrogen for 2 hours. After completion, the reaction mixture was cooled to room temperature and purified by silica gel column chromatography (PE:EA = 1:3) to obtain compound I005f (1.00 g, yield 66.8%). MS m / z (ESI): 390.2 [M+H] + .

[0287] Step 6:

[0288] (R)-N-{(S)-{1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methyl}-2-methylpropane-2-sulfenamide (1005 g)

[0289] Compound I005f (1.72 g, 4.42 mmol) was dissolved in anhydrous DCM (40 mL), the atmosphere was replaced with nitrogen, and a solution of lithium tri-sec-butylborohydride (1 M in THF, 8.9 mL, 8.84 mmol) was added at 0°C. The reaction system was stirred at 0°C for 2 hours. After completion of the reaction, saturated ammonium chloride (10 mL) was added to quench the reaction. The solid precipitated and was filtered, and the filtrate was extracted with DCM (30 mL x 3). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (DCM:MeOH = 100:1) to obtain compound I005g (1.30 g, yield 75.2%) as a white solid. MS m / z (ESI): 392.2 [M+H] + .

[0290] Step 7:

[0291] (S)-{1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methanamine (I005)

[0292] At room temperature, compound I005g (1.30g, 3.32mmol) was dissolved in anhydrous DCM (10mL). A solution of hydrogen chloride in dioxane (1mL, 4M) was added and stirred for 2 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to remove the solvent. The pH was adjusted to a weak base using saturated sodium bicarbonate and concentrated under reduced pressure to obtain the crude product. This was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound I005 as a white solid (900mg, 94.4% yield). MS m / z (ESI): 288.1 [M+H] +.

[0293] Intermediate 6

[0294] Preparation of (S)-[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methanamine (I006)

[0295] Step 1:

[0296] Methyl [1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)acetate (I006a)

[0297] Compound I004 (3 g, 14.5 mmol), copper sulfate pentahydrate (1.8 g, 7.25 mmol), and sodium ascorbate (1.43 g, 7.25 mmol) were added to 2-methyltetrahydrofuran (30 mL). A solution of I002b in acetonitrile (29 mmol, 35 mL, approximately 0.85 M) was then added. The mixture was reacted at 75°C under nitrogen for 24 hours. After completion, the reaction solution was concentrated, and DCM (100 mL) and H₂O (50 mL) were added. The mixture was filtered, and the filtrate was separated. The aqueous phase was extracted twice with DCM. The combined organic layers were washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (PE:EA = 1:1) afforded compound I006a (2.95 g, 64.3% yield) as a yellow solid. MS m / z (ESI): 317.0 [M+H] + .

[0298] Step 2:

[0299] [1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methanol (I006b)

[0300] Compound I006a (2.95 g, 9.33 mmol) was dissolved in THF (30 mL), and a solution of sodium hydroxide (746 mg, 18.6 mmol) in water (10 mL) was added. The mixture was allowed to react at 20°C for 12 hours. After completion, the pH was adjusted to 5-6 with dilute HCl. The THF was concentrated to remove the aqueous layer, and the aqueous layer was extracted with DCM (3 x 20 mL). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to afford Compound I006b (2.2 g, 86.0% yield) as a yellow solid. MS m / z (ESI): 275.1 [M+H] + .

[0301] Step 3:

[0302] [1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methanone (I006c)

[0303] Compound I006b (2.2 g, 8.03 mmol) was dissolved in anhydrous DCM (40 mL), and Dess-Martin reagent (5.1 g, 12.05 mmol) was added portionwise. The mixture was allowed to react at 30°C for 4 hours. After completion, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted twice with DCM. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (PE:EA = 2:1) to afford compound I006c (2.0 g, 91.5% yield). MS m / z (ESI): 273.1 [M+H] + .

[0304] Step 4:

[0305] (S,E)-N-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methylene}-2-methylpropane-2-sulfonamide (I006d)

[0306] Compound I006c (2.0 g, 7.35 mmol) and (R)-(+)-tert-butylsulfenamide (2.66 g, 22.0 mmol) were dissolved in anhydrous toluene (20 mL) under nitrogen atmosphere. Tetraethyl titanate (5.02 g, 22.0 mmol) was added and allowed to react at 75°C for 16 hours. After completion of the reaction, the temperature was cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound I006d (1.5 g, 54.4% yield) as a white solid. MS m / z (ESI): 376.1 [M+H] + .

[0307] Step 5:

[0308] (S)-N-{(S)-[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methyl}-2-methylpropane-2-sulfonamide (I006e)

[0309] Compound I006d (1.5 g, 4 mmol) was dissolved in anhydrous DCM (20 mL) under nitrogen protection, with the temperature maintained below 0°C. A solution of lithium tri-sec-butylborohydride in tetrahydrofuran (1.0 N, 8 mL, 8 mmol) was slowly added dropwise. The reaction was allowed to react at 0°C for 4 hours. After completion, saturated aqueous ammonium chloride was slowly added dropwise to quench the reaction. The aqueous layer was extracted twice with DCM. The combined organic layers were washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford Compound I006e (1.3 g, 86.2% yield) as a yellow solid. MS m / z (ESI): 378.1 [M+H] + .

[0310] Step 6:

[0311] (S)-[1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methanamine (I006)

[0312] Compound I006e (1.3 g, 3.45 mmol) was dissolved in DCM (15 mL) and a dioxane hydrochloride solution (4.0 N, 2 mL) was added dropwise. The mixture was allowed to react at 20°C for 2 hours. After completion, the reaction mixture was concentrated, and the crude product was diluted with water (10 mL). The pH was adjusted to approximately 7 and then concentrated. The crude product was purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound I006 (650 mg, 69% yield). MS m / z (ESI): 274.1 [M+H] + .

[0313] Intermediate 7

[0314] Preparation of (S)-5-{amino[1-(1-(difluoromethyl)cyclopropyl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I007)

[0315] Step 1:

[0316] Methyl {1-[1-(difluoromethyl)cyclopropyl]-1H-1,2,3-triazol-4-yl}(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)acetate (I007a)

[0317] At room temperature, I001 (1.0 g, 3.92 mmol), copper sulfate pentahydrate (490 mg, 1.96 mmol), and sodium ascorbate (380 mg, 1.96 mmol) were added to 2-methyltetrahydrofuran (20 mL). A solution of 1-azido-1-(difluoromethyl)cyclopropane (5.88 mmol) in acetonitrile was then added. The reaction mixture was stirred at 75°C under nitrogen for 12 hours. After completion, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain the crude product. The product was diluted with water (20 mL) and extracted with EA (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The product was purified by silica gel column chromatography (DCM:MeOH = 3:1) to afford compound I007a (600 mg, 39.5% yield). MS m / z (ESI): 389.1 [M+H] + .

[0318] Step 2:

[0319] 5-{{{1-[1-(Difluoromethyl)cyclopropyl]-1H-1,2,3-triazol-4-yl}hydroxy}methyl}-2-methylisoquinolin-1(2H)-one (I007b)

[0320] At room temperature, compound I007a (600 mg, 1.55 mmol) was dissolved in THF (20 mL). A solution of lithium hydroxide monohydrate (195 mg, 4.65 mmol) in water (2 mL) was added, and the reaction mixture was stirred at room temperature overnight. After completion, the pH of the solution was adjusted to 7-8 using 1.0 N hydrochloric acid. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 3:1) to obtain compound I007b (500 mg, 93.2% yield) as a yellow solid. MS m / z (ESI): 347.0 [M+H] + .

[0321] Step 3:

[0322] 5-{1-[1-(difluoromethyl)cyclopropyl]}-1H-1,2,3-triazole-4-carbonyl-2-methylisoquinolin-1(2H)-one (I007c)

[0323] Compound I007b (500 mg, 1.4 mmol) was dissolved in anhydrous DCM (10 mL) at room temperature, and Dess-Martin reagent (1.2 g, 2.8 mmol) was added. The reaction mixture was stirred at room temperature overnight. After completion of the reaction, the reaction mixture was filtered, and the filtrate was quenched with saturated aqueous sodium bicarbonate (10 mL) and extracted with DCM (20 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. Purification by silica gel column chromatography (PE:EA = 1:2) afforded compound I007c (350 mg, 72.9% yield) as a yellow solid. MS m / z (ESI): 345.1 [M+H] + .

[0324] Step 4:

[0325] (R,E)-N-{{1-[1-(difluoromethyl)cyclopropyl]-1H-1,2,3-triazol-4-yl}(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)methylene}-2-methylpropane-2-sulfonamide (I007d)

[0326] Compound I007c (350 mg, 1.0 mmol) was dissolved in tetraethyl titanate (10 mL), and (R)-2-methylpropane-2-sulfonamide (360 mg, 3.0 mmol) was added. The reaction mixture was stirred at 75°C overnight under nitrogen. After completion, the reaction solution was cooled to room temperature, and DCM (10 mL) was added. The product was concentrated to obtain a crude product, which was purified by silica gel column chromatography (PE:EA = 1:3) to obtain compound I007d (330 mg, 73.8% yield) as a yellow solid. MS m / z (ESI): 448.2 [M+H] + .

[0327] Step 5:

[0328] (R)-N-{{(S)-1-[1-(difluoromethyl)cyclopropyl]-1H-1,2,3-triazol-4-yl}(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)methyl}-2-methylpropane-2-sulfonamide (I007e)

[0329] Compound I007d (330 mg, 0.74 mmol) was dissolved in anhydrous DCM (10 mL), the atmosphere was purged with nitrogen, and a solution of lithium tri-sec-butylborohydride (1 M in THF, 1.5 mL, 1.48 mmol) was added at 0°C. The reaction system was stirred at 0°C for 2 hours. After completion, saturated ammonium chloride (10 mL) was added to quench the reaction. A solid precipitated and was filtered. The filtrate was extracted with DCM (30 mL). The organic phases were combined, dried, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound I007e (280 mg, 84.3% yield) as a yellow oil. MS m / z (ESI): 450.0 [M+H] + .

[0330] Step 6:

[0331] (S)-5-{Amino[1-(1-(difluoromethyl)cyclopropyl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I007)

[0332] Compound I007e (280 mg, 0.62 mmol) was dissolved in DCM (10 mL), and a solution of hydrochloric acid in dioxane (1 mL, 4 M) was added. The mixture was stirred for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove the organic solvent. The pH was adjusted to a weak base using saturated sodium bicarbonate. The crude product was then concentrated under reduced pressure. This was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound I007 (200 mg, 93.8% yield) as a yellow solid. MS m / z (ESI): 691.2 [2M+H] + .

[0333] Intermediate 8

[0334] Preparation of 6-bromo-8-chloro-N-(3,3-dimethyltetrahydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidin-4-amine (I008)

[0335] Step 1:

[0336] Compound I003e (300 mg, 1.15 mmol), 3,3-dimethyloxan-4-amine (600 mg, 4.61 mmol), and triethylamine (460 mg, 4.61 mmol) were dissolved in DMF (15 mL) and stirred at 0°C under nitrogen for 30 minutes. PyBOP (1.2 g, 2.3 mmol) was added, and the mixture was stirred at 0°C for 2 hours. After completion of the reaction, the mixture was diluted with H2O (20 mL) and extracted with EA (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (PE:EA = 1:1) to obtain Compound I008 (150 mg, 35.2% yield) as a yellow solid. MS m / z (ESI): 371.0 / 373.0 [M+H] + .

[0337] Intermediate 9

[0338] 5-{(1S)-amino[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I009)

[0339] Step 1:

[0340] Methyl (2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]acetate (I009c)

[0341] Compound I009a (1.0 g, 8.3 mmol) was dissolved in acetonitrile (10 mL), and DIPEA (2.35 g, 18.2 mmol) was added. The mixture was stirred for 10 minutes. AMDP (3.07 g, 10.8 mmol) was added in portions under nitrogen protection, and the mixture was stirred at 25°C for 24 hours. Compound I001 (1.6 g, 6.3 mmol), copper sulfate pentahydrate (784 mg, 3.15 mmol), sodium ascorbate (623 mg, 3.15 mmol), and 2-methyltetrahydrofuran (40 mL) were then added, and the mixture was reacted at 75°C for 24 hours under nitrogen. The mixture was then concentrated under reduced pressure, and DCM (100 mL) and water (50 mL) were added. The mixture was filtered, and the filtrate was separated. The aqueous phase was extracted twice with DCM. The organic layers were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound I009c (1.1 g, 36% yield). MS m / z (ESI): 365.1 [M+H] + .

[0342] Step 2:

[0343] 5-{Hydroxy[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I009d)

[0344] Compound I009c (1.1 g, 3.0 mmol) was dissolved in THF (10 mL) and H₂O (5 mL), and lithium hydroxide monohydrate (240 mg, 6 mmol) was added. The mixture was allowed to react at 20°C for 12 hours. After completion, the pH was adjusted to 5 with dilute HCl and concentrated under reduced pressure. The aqueous layer was extracted three times with DCM. The combined organic layers were washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to afford Compound I009d (780 mg, 80.0% yield) as a yellow solid. MS m / z (ESI): 323.1 [M+H] + .

[0345] Step 3:

[0346] 2-Methyl-5-[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazole-4-carbonyl]isoquinolin-1(2H)-one (I009e)

[0347] Compound I009d (780 mg, 2.4 mmol) was dissolved in anhydrous DCM (20 mL), and Dess-Martin reagent (2.0 g, 4.8 mmol) was added portionwise. The mixture was allowed to react at 30°C for 4 hours. After completion, the reaction was washed twice with saturated sodium bicarbonate solution. The aqueous layer was extracted twice with DCM. The organic layers were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE:EA = 2:1) to afford compound I009e (550 mg, 71% yield). MS m / z (ESI): 321.1 [M+H] + .

[0348] Step 4:

[0349] (R)-2-Methyl-N-{(E)-(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methylene}propane-2-sulfenamide (I009f)

[0350] Compound I009e (550 mg, 1.71 mmol) and (R)-(+)-tert-butylsulfenamide (623 mg, 5.15 mmol) were added to anhydrous toluene (10 mL), followed by tetraethyl titanate (1.17 g, 5.15 mmol). Under nitrogen, the mixture was reacted at 75°C for 16 hours. After completion, the reaction was cooled to room temperature, silica gel was added, and the mixture was concentrated. The mixture was purified by silica gel column chromatography (PE:EA = 1:3) to obtain compound I009f (350 mg, 48% yield). MS m / z (ESI): 424.1 [M+H] + .

[0351] Step 5:

[0352] (R)-2-Methyl-N-{(1S)-(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}propane-2-sulfenamide (1009 g)

[0353] Compound I009f (350 mg, 0.83 mmol) was dissolved in anhydrous DCM (10 mL) under nitrogen atmosphere. The temperature was lowered to -0°C, and a solution of lithium tri-sec-butylborohydride in tetrahydrofuran (1.65 mL, 1.65 mmol, 1.0 mol / L) was slowly added dropwise. The mixture was allowed to react at 0°C for 4 hours. After completion, saturated aqueous ammonium chloride was added dropwise to quench the reaction. The aqueous layer was extracted twice with DCM. The combined organic layers were washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound I009g (290 mg, 82% yield). MS m / z (ESI): 426.1 [M+H] + .

[0354] Step 6:

[0355] 5-{(1S)-amino[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (I009)

[0356] Compound I009g (290 mg, 0.68 mmol) was dissolved in DCM (10 mL), and a 4.0 M HCl / 1.4-dioxane solution (3 mL) was added. The mixture was allowed to react at 25°C for 4 hours. After completion of the reaction, the pH of the reaction solution was adjusted to approximately 7 with saturated sodium bicarbonate solution. The mixture was extracted with DCM (20 mL x 3). The organic layers were combined, washed with saturated sodium chloride solution (20 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (MeOH:DCM = 0-10%) to obtain compound I009 (160 mg, 73% yield). MS m / z (ESI): 643.3 [2M+H]+ .

[0357] Intermediate 10

[0358] Preparation of 6-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (I010)

[0359] Step 1:

[0360] 3-Amino-2,6-dichloroisonicotinamide (I010b)

[0361] Compound I010a (4 g, 18.10 mmol) was dissolved in 7.0 M ammonia-methanol solution (80 mL) and stirred at room temperature for 2 days. After completion of the reaction, the mixture was concentrated to give the crude product. EA (15 mL) was added, stirred at 25°C for 10 minutes, and filtered to give I010b (3.70 g, 99% yield), a white solid. MS m / z (ESI): 207.0 [M+H] + .

[0362] Step 2:

[0363] 3-Amino-2,6-dichloro-N-[(dimethylamino)methylene]isonicotinamide (I010c)

[0364] Compound I010b (3.70 g, 17.96 mmol) was dissolved in 1,4-dioxane (30 mL), and DMF-DMA (3.23 g, 27.53 mmol) was slowly added. The mixture was stirred at 80°C under a nitrogen atmosphere for 1 hour. After the reaction was complete, the mixture was cooled and concentrated to obtain the crude yellow solid product I010c (3.60 g, 76.77% yield), which was used directly in the next step. MS m / z (ESI): 262.1 [M+H] + .

[0365] Step 3:

[0366] 6,8-Dichloropyrido[3,4-d]pyrimidin-4-ol (I010d)

[0367] Compound I010c (3.6 g, 13.79 mmol) was dissolved in 1,4-dioxane (40 mL) at room temperature, and cesium carbonate (13.48 g, 41.37 mmol) was added with stirring. The reaction mixture was stirred at 100°C for 2 hours. After completion, the reaction was diluted with water (50 mL) and extracted with EA (30 mL x 3). The organic phases were combined, washed with water (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound I010d (2.8 g, 94% yield) as a white solid. MS m / z (ESI): 217.0 [M+1] + .

[0368] Step 4:

[0369] 6,8-Dichloro-N-neopentylpyrido[3,4-d]pyrimidin-4-amine (I010e)

[0370] Compound I010d (843 mg, 3.90 mmol), neopentylamine (1.36 g, 15.61 mmol), and DIPEA (1.51 g, 11.70 mmol) were dissolved in DMF (15 mL) and stirred at 0°C under nitrogen for 30 minutes. PyBOP (4.06 g, 7.80 mmol) was added, and the reaction was continued at 0°C with stirring for 2 hours. After completion, the reaction was diluted with water (20 mL) and extracted with EA (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (PE:EA = 1:1) to obtain the white solid product I010e (70 mg, yield 6.41%). MS m / z (ESI): 286.0 [M+H] + .

[0371] Step 5:

[0372] 6-Chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (I010)

[0373] At room temperature, I010e (310 mg, 1.64 mmol) and zinc cyanide (96.18 mg, 0.82 mmol) were dissolved in dioxane (8 mL). XantPhos Pd G2 (142.19 mg, 0.16 mmol) was added, and the reaction mixture was stirred at 120°C under nitrogen for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (MeOH:DCM = 0-15%) to obtain compound I010 (210 mg, 46% yield). MS m / z (ESI): 276.1 [M+H] + .

[0374] Intermediate 11

[0375] 8-Chloro-N 4 -neopentylpyrido[3,4-d]pyrimidine-4,6-diamine (I011)

[0376] Step 1:

[0377] Tert-butyl [8-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl]carbamate (I011a)

[0378] Compound I003 (2 g, 6.09 mmol), NH2Boc (748 mg, 6.39 mmol), Cs2CO3 (3.95 g, 12.18 mmol), and XantPhos PdG2 (532 mg, 0.6 mmol) were dissolved in 1,4-dioxane (20 mL) and stirred at 90°C under nitrogen for 6 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:CH3OH = 0-10%) to obtain compound I011a (1.7 g, 76% yield). MS m / z (ESI): 366.1 [M+H] + .

[0379] Step 7:

[0380] 8-Chloro-N 4 -neopentylpyrido[3,4-d]pyrimidine-4,6-diamine (I011)

[0381] Compound I011a (1.7 g, 4.65 mmol) was dissolved in DCM (20 mL), followed by the addition of TFA (20 mL). The mixture was stirred at 20°C under nitrogen for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, diluted with DCM (30 mL) and H₂O (30 mL), and adjusted to pH 7 with saturated sodium bicarbonate solution. The mixture was extracted with DCM (50 mL x 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (DCM:EA = 1:1) to obtain compound I011 (0.95 g, 77% yield). MS m / z (ESI): 266.1 [M+H] + .

[0382] Intermediate 12

[0383] Preparation of 1-(8-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)prop-2-yn-1-yl acetate (I012)

[0384] Step 1:

[0385] 5-Formyl-2-methylisoquinolin-2-ium (I012b)

[0386] To a 250 mL single-necked flask, 5-bromo-8-fluoroisoquinoline (5.00 g, 22.11 mmol) and acetonitrile (40 mL) were added. The temperature was raised to 30°C, and iodomethane (2.75 mL, 44.23 mmol) was added. The mixture was stirred at 30°C for 18 hours. Toluene (10 mL) was added, stirred, and filtered. The filter cake was rinsed with toluene (10 mL) and dried to afford compound I012b (5.00 g, 94.37% yield).

[0387] Step 2:

[0388] 5-Bromo-8-fluoro-2-methylisoquinolin-1(2H)-one (I012c)

[0389] To a 50 mL single-necked vial, compound I012b (5.00 g, 20.74 mmol), DBU (4.68 mL, 31.11 mmol), and THF (100 mL) were added, followed by pentacyclic trimethylphenylhydrazine triazole (1.07 g, 3.40 mmol). Stirring was continued at 25°C under oxygen protection for 18 hours. After completion, the reaction was quenched with water and extracted with EA. The organic phase was concentrated and purified by silica gel column chromatography (PE:EA = 8:1) to afford compound I012c (5.00 g, 94.15% yield).

[0390] Step 3:

[0391] 8-Fluoro-2-methyl-5-vinylisoquinolin-1(2H)-one (I012d)

[0392] Compound I012c (5.00 g, 19.52 mmol), potassium vinyl fluoroborate (3.95 g, 29.28 mmol), Pd(dppf)Cl2 (0.71 g, 0.97 mmol), and H2O (10 mL) were dissolved in dioxane (50 mL). Potassium carbonate (8.10 g, 58.57 mmol) was added, and the mixture was reacted at 100°C under nitrogen for 18 hours. After completion, H2O (100 mL) was added to the reaction solution, which was extracted with DCM. The organic phase was concentrated and purified by silica gel column chromatography (PE:EA = 5:1) to afford compound I012d (3.10 g, 78.21% yield).

[0393] Step 4:

[0394] 8-Fluoro-2-methyl-1-oxo-1,2-dihydroisoquinoline-5-carbaldehyde (I012e)

[0395] Compound I012d (3.10 g, 15.25 mmol), H₂O (10 mL), and potassium osmate (0.12 g, 0.38 mmol) were dissolved in THF (40 mL). Sodium periodate (9.79 g, 45.76 mmol) was added and reacted at 25°C under nitrogen for 18 hours. The reaction solution was poured into water (100 mL) and extracted with EA (50 mL x 3). The combined organic phases were washed with water (50 mL) and then with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE:EA = 1:1) to afford compound I012e (1.92 g, 61.34% yield) as a white solid.

[0396] Step 5:

[0397] 8-Fluoro-5-(1-hydroxy-2-yn-1-yl)-2-methylisoquinolin-1(2H)-one (I012f)

[0398] Compound I012e (1.91 g, 9.26 mmol) was dissolved in THF (20 mL), and ethynylmagnesium chloride (13.89 mL, 13.89 mmol) was added at 0°C. The mixture was reacted under nitrogen at 0°C for 1 hour. After completion, the reaction solution was poured into water (100 mL) and extracted with EA (50 mL x 3). The organic phases were combined, washed with water (50 mL), then saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered to yield compound I012f (1.60 g, 59.78% yield).

[0399] Step 6:

[0400] 1-(8-Fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)prop-2-yn-1-yl acetate (I012)

[0401] Compound I012f (1.60 g, 5.53 mmol) and triethylamine (3.46 mL, 24.91 mmol) were added to DCM (20 mL). Acetyl chloride (1.18 mL, 16.61 mmol) was added at 0°C. The reaction was allowed to proceed for 0.5 hours at 0°C, then the temperature was raised to 25°C and the reaction continued for 18 hours. The reaction solution was quenched with water (200 mL) and extracted with EA. The organic phase was concentrated and purified by silica gel column chromatography (PE:EA = 3:1) to afford Compound I012 (1.41 g, 92.55% yield) as a white solid.

[0402] Intermediate 13

[0403] Preparation of 1-(1-methyl-2-oxo-1,2-dihydroquinolin-5-yl)prop-2-yn-1-yl acetate (I013)

[0404] Step 1:

[0405] 5-Formyl-1-methylquinolin-1-ium (I013b)

[0406] Compound I013a (4.00 g, 25.5 mmol) was dissolved in acetonitrile (25 mL), and iodomethane (0.746 M, 10 mL) was added. After the addition was complete, the reaction mixture was stirred at room temperature under nitrogen protection overnight. After completion of the reaction, the mixture was filtered, and the filter cake was rinsed with toluene and dried to afford Compound I013b (6.70 g, 87.9% yield) as an orange solid. MS m / z (ESI): 172.2 [M+H] + Step 2:

[0407] 1-Methyl-2-oxo-1,2-dihydroquinoline-5-carbaldehyde (I013c)

[0408] Compound I013b (6.70 g, 22.4 mmol) and potassium hexacyanoferrate (14.7 g, 44.8 mmol) were dissolved in H2O (80 mL). Under nitrogen, a 20 mL aqueous solution of potassium hydroxide (5.02 g) was added dropwise at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 2 hours. After the reaction was complete, EA (100 mL x 3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was separated and purified by silica gel column chromatography (PE:EA = 1:3) to obtain compound I013c (1.5 g, 35.8% yield) as a white solid. MS m / z (ESI): 188.1 [M+H] + .

[0409] Step 3:

[0410] 5-(1-Hydroxy-2-yn-1-yl)-1-methylquinolin-2(1H)-one (I013d)

[0411] Compound I013c (1.50 g, 8.02 mmol) was dissolved in THF (10 mL). Under nitrogen, ethynylmagnesium bromide (0.746 M, 10 mL) was added dropwise at 0°C. After addition, the reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction was quenched with a saturated aqueous solution of ammonium chloride (10 mL). DCM (20 mL x 3) was added for extraction. The organic phases were combined, dried over anhydrous ammonium sulfate, and filtered. The filtrate was concentrated under reduced pressure to yield compound I013d (1.87 g, crude product). MS m / z (ESI): 214.1 [M+H] + .

[0412] Step 4:

[0413] 1-(1-Methyl-2-oxo-1,2-dihydroquinolin-5-yl)prop-2-yn-1-yl acetate (I013e)

[0414] Compound I013d (1.87 g, 8.78 mmol), 4-dimethylaminopyridine (107 mg, 0.878 mmol), and triethylamine (2.66 g, 26.3 mmol) were dissolved in DCM (20 mL). Under nitrogen, acetic anhydride (1.80 g, 17.6 mmol) was added dropwise at 0°C. After addition, the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, H2O (20 mL) was added and extracted with DCM (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (EA:PE = 1:2) to afford compound I013e (1.9 g, 84.6% yield) as a white solid. MS m / z (ESI): 256.0 [M+H] + Step 5:

[0415] Methyl [1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](1-methyl-2-oxo-1,2-dihydroquinolin-5-yl)acetate (I013f)

[0416] Compound I013e (1.90 g, 7.45 mmol), 1-azidobicyclo[1.1.1]pentane (100 mg / mL, 13 mL, 11.2 mmol), copper sulfate pentahydrate (933 mg, 3.73 mmol), and sodium ascorbate (739 mg, 3.73 mmol) were dissolved in 2-methyltetrahydrofuran (30 mL). After addition, the reaction mixture was stirred at 75°C under nitrogen overnight. After cooling to room temperature, water (20 mL) was added and extracted with EA (30 mL x 3). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (DCM:CH3OH = 50:1) to afford Compound I013f (2.20 g, 80.9% yield) as a brown oil. MS m / z (ESI): 365.2 [M+H] + .

[0417] Step 6:

[0418] 5-{[1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](hydroxy)methyl}-1-methylquinolin-2(1H)-one (1013g)

[0419] At room temperature, compound I013f (2.20 g, 6.04 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL), and an aqueous solution (4 mL) of lithium hydroxide monohydrate (508 mg, 12.1 mmol) was added dropwise to the reaction solution. The reaction was stirred at room temperature for 3 hours. The reaction mixture was adjusted to pH 7-8 with 1M hydrochloric acid solution. Water (10 mL) was added and extracted with EA (30 mL). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (DCM:CH3OH=1:20) to obtain compound I013g (1.91 g, yield 98.0%) as a yellow oil. MS m / z (ESI): 323.2 [M+H] + .

[0420] Step 7:

[0421] 5-[1-(Bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazole-4-carbonyl]-1-methylquinolin-2(1H)-one (I013h)

[0422] Compound I013g (1.91 g, 5.93 mmol) was dissolved in anhydrous DCM (20 mL), and Dess-Martin periodinane (5.05 g, 11.9 mmol) was added. The reaction system was stirred at room temperature overnight. Filtered, the filtrate was quenched with saturated aqueous sodium bicarbonate solution (10 mL), and extracted with DCM (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (PE:EA = 2:1) to obtain compound I013h (1.80 g, yield 93.7%) as a yellow solid. MS m / z (ESI): 321.1 [M+H] + .

[0423] Step 8:

[0424] (S,E)-N-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](1-methyl-2-oxo-1,2-dihydroquinolin-5-yl)methylene}-2-methylpropane-2-sulfamide (I013i)

[0425] Compound I013h (1.80 g, 5.63 mmol) was dissolved in toluene (20 mL), and tetraethyl titanate (5.13 g, 22.5 mmol) and (S)-2-methylpropane-2-sulfonamide (2.04 g, 16.9 mmol) were added. The reaction mixture was stirred at 75°C under nitrogen for 16 hours. The reaction mixture was cooled to room temperature and purified by silica gel column chromatography (PE:EA = 1:4) to obtain compound I013i (1.50 g, 63.0% yield) as a yellow solid. MS m / z (ESI): 424.2 [M+H] + .

[0426] Step 9:

[0427] (S)-N-{(S)-[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](1-methyl-2-oxo-1,2-dihydroquinolin-5-yl)methyl}-2-methylpropane-2-sulfamide (I013j)

[0428] Compound I013i (1.50 g, 3.54 mmol) was dissolved in anhydrous DCM (40 mL), the atmosphere was replaced with nitrogen, and a solution of lithium tri-tert-butylaluminum hydride (1 M / THF, 7.1 mL, 7.08 mmol) was added at 0°C. The reaction system was stirred at 0°C for 2 hours. Saturated ammonium chloride (10 mL) was added to quench the mixture, and a solid precipitated. The solid was filtered, and the filtrate was extracted with DCM (30 mL x 3). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to obtain compound I013j (1.30 g) as a white solid, which was used in the next step without further purification. MS m / z (ESI): 426.2 [M+H] + .

[0429] Step 10:

[0430] (S)-5-{amino[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-1-methylquinolin-2(1H)-one (I013)

[0431] Compound I013j (1.50 g, 3.52 mmol) was dissolved in anhydrous DCM (20 mL) at room temperature, and a solution of hydrogen chloride in dioxane (2 mL, 4 M) was added. The mixture was stirred and reacted for 2 hours. The reaction solution was concentrated under reduced pressure to remove the organic solvent, and the pH was adjusted to a weak base using saturated sodium bicarbonate. The crude product was then concentrated under reduced pressure to obtain the product, which was purified by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound I013 (500 mg, 44.2% yield) as a yellow solid. MS m / z (ESI): 322.1 [M+H] + .

[0432] Intermediate 14

[0433] Preparation of 1-([1,2,4]triazolo[3,4-a]isoquinolin-7-yl)prop-2-yn-1-yl acetate (I014)

[0434] Step 1:

[0435] (Z)-5-Bromo-1-hydrazino-1,2-dihydroisoquinoline (I014b)

[0436] Compound I014a (10.00 g, 41.23 mmol) was dissolved in EtOH (100 mL). Hydrazine hydrate (7.58 mL, 123.71 mmol) was added to the reaction mixture, and the mixture was stirred at 80°C for 18 hours. After completion of the reaction, the reaction mixture was spin-dried to obtain crude product I014b (9.80 g, 99.82% yield) as an orange solid, which was used in the next step without further purification. MS m / z (ESI): 240.2 [M+H] + .

[0437] Step 2:

[0438] 7-Bromo-[1,2,4]triazolo[3,4-a]isoquinoline (I014c)

[0439] Compound I014b (9.80 g, 41.16 mmol) was dissolved in DMF (100 mL). Imidazole hydrochloride (0.43 g, 4.11 mmol) was added to the reaction mixture and reacted at 135°C for 18 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (DCM:CH3OH = 20:1) to obtain compound I014c (10.2 g, 99.89% yield) as a white solid. MS m / z (ESI): 250.2 [M+H] + .

[0440] Step 3:

[0441] 7-vinyl-[1,2,4]triazolo[3,4-a]isoquinoline (I014d)

[0442] Compound I014c (8.50 g, 34.26 mmol) was dissolved in a mixture of 1,4-dioxane (100 mL) and H₂O (20 mL). Potassium vinyl trifluoroborate (6.88 g, 51.39 mmol), potassium carbonate (14.21 g, 102.78 mmol), and Pd(dppf)Cl₂ (1.25 g, 1.71 mmol) were added to the reaction mixture. The mixture was reacted at 100°C under nitrogen for 18 hours. After completion, the reaction mixture was concentrated and purified by silica gel column chromatography (DCM:CH₃OH = 10:1) to afford compound I014d (6.50 g, 97.18% yield) as a white solid. MS m / z (ESI): 196.2 [M+H] + .

[0443] Step 4:

[0444] [1,2,4]triazolo[3,4-a]isoquinoline-7-carboxylic acid (I014e)

[0445] Compound I014d (5.00 g, 25.61 mmol) was dissolved in a mixture of THF (40 mL) and H₂O (10 mL). Sodium periodate (16.43 g, 76.83 mmol) and potassium(VI) osmate (0.47 g, 1.28 mmol) were added to the reaction solution, and the reaction mixture was allowed to react at 25°C for 18 hours. After completion, the reaction solution was concentrated and purified by silica gel column chromatography (DCM:CH₃OH=10:1) to obtain compound I014e (5.00 g, yield 99.01%) as a white solid. MS m / z (ESI): 198.2 [M+H] + .

[0446] Step 5:

[0447] 1-([1,2,4]triazolo[3,4-a]isoquinolin-7-yl)prop-2-yn-1-ol (I014f)

[0448] Compound I014e (4.50 g, 22.81 mmol) was dissolved in THF (40 mL). The reaction mixture was cooled to 0°C and ethynylmagnesium chloride (34.22 mL, 34.22 mmol) was added. The reaction mixture was allowed to react at 0°C for 1 hour. The reaction mixture was slowly poured into H2O (100 mL) and extracted with EA (20 mL x 3). The organic phases were combined, washed with H2O (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered to dryness to obtain crude product I014f (4.50 g, 98.01% yield) as a yellow solid. The crude product was used directly in the next step. MS m / z (ESI): 224.2 [M+H] + .

[0449] Step 6:

[0450] 1-([1,2,4]triazolo[3,4-a]isoquinolin-7-yl)prop-2-yn-1-yl acetate (I014)

[0451] Compound I014f (4.50 g, 20.15 mmol) was dissolved in DMF (30 mL). Triethylamine (8.40 mL, 60.47 mmol) was added to the reaction solution. The reaction solution was cooled to 0°C and acetyl chloride (2.86 mL, 40.31 mmol) was added. The reaction was allowed to react at 0°C for 2 hours. The reaction solution was concentrated and purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound I014 (1.00 g, yield 21.04%) as a yellow solid. MS m / z (ESI): 266.2 [M+H] + .

[0452] Intermediate 15

[0453] Preparation of 1-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)prop-2-yn-1-yl acetate (I015)

[0454] Step 1:

[0455] 8-Bromo-2H-benzo[b][1,4]oxazin-3(4H)-one (I015b)

[0456] At room temperature, compound I015a (6.0 g, 32.0 mmol) and K2CO3 (11.0 g, 80.0 mmol) were dissolved in acetonitrile (150 mL) and water (150 mL). Chloroacetyl chloride (5.4 g, 48.0 mmol) was added dropwise under nitrogen and cooled to 0°C. After the addition, the reaction mixture was heated to 100°C and stirred for 16 hours. After the reaction was complete, water (100 mL) was added and extracted with EA (300 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (PE:EA = 3:1) to afford I015b (4.5 g, 61.6% yield) as a brown solid. MS m / z (ESI): 228.0 [M+H] + .

[0457] Step 2:

[0458] 8-Vinyl-2H-benzo[b][1,4]oxazin-3(4H)-one (I015c)

[0459] Compound I015b (4.5 g, 19.7 mmol), potassium carbonate (8.2 g, 59.1 mmol), potassium trifluoro(vinyl)borate (3.9 g, 30.0 mmol), and Pd(dppf)Cl2 (1.4 g, 2.0 mmol) were dissolved in 1,4-dioxane (100 mL) and water (10 mL) at room temperature. The reaction mixture was stirred at 100°C under nitrogen for 10 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE:EA = 1:5) to afford I015c (3.0 g, 88.2% yield) as a yellow solid. MS m / z (ESI): 176.1 [M+H] + .

[0460] Step 3:

[0461] 3-Oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-8-carboxylic acid (I015d)

[0462] Compound I015c (3.0 g, 17.1 mmol) and potassium osmate dihydrate (630.0 mg, 1.7 mmol) were dissolved in tetrahydrofuran (100 mL) and water (10 mL). The mixture was stirred at 0°C for 30 min, followed by the addition of sodium periodate (10.9 g, 51.3 mmol). The reaction was stirred at 0°C overnight. After the reaction was complete, water (50 mL) was added, and the mixture was extracted with EA (200 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (PE:EA = 2:1) to afford I015d (2.0 g, 66.7% yield) as a yellow solid. MS m / z (ESI): 178.1 [M+H] + .

[0463] Step 4:

[0464] 8-(1-Hydroxy-2-yl-1-yl)-2H-benzo[b][1,4]oxazin-3(4H)-one (I015e)

[0465] Compound I015d (2.0 g, 11.3 mmol) was dissolved in tetrahydrofuran (50 mL), the atmosphere was replaced with nitrogen, and ethynylmagnesium bromide (80 mL, 40.0 mmol, 0.5 M / THF) was added at 0°C. The mixture was stirred for 2 hours. After the reaction was complete, water (50 mL) was added and extracted with EA (200 mL). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to afford crude product I015e (1.3 g, 56.5% yield). MS m / z (ESI): 204.1 [M+H] + .

[0466] Step 5:

[0467] 1-(3-Oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)prop-2-yn-1-yl acetate (I015)

[0468] Compound I015e (1.3 g, 6.4 mmol) was dissolved in anhydrous dichloromethane (20 mL), and 4-dimethylaminopyridine (80.0 mg, 0.64 mmol), triethylamine (1.9 g, 19 mmol), and acetic anhydride (1.3 g, 12.8 mmol) were added. The reaction mixture was stirred at 0°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE:EA = 1:1) to obtain I015 as a yellow solid (1.0 g, yield 62.5%). MS m / z (ESI): 246.1 [M+H] + .

[0469] Intermediate 16

[0470] Preparation of 5-{1-{[8-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl]amino}propyl-2-yn-1-yl}-2-methylphthalazin-1(2H)-one (I016)

[0471] Step 1:

[0472] 4-Bromo-3-hydroxyisobenzofuran-1(3H)-one (I016b)

[0473] 2,2,6,6-Tetramethylpiperidine (44.1 g, 313 mmol) was dissolved in tetrahydrofuran (150 mL). n-Butyl lithium (125 mL, 313 mmol) was added dropwise at -60°C, and the reaction was stirred for 1 hour. A solution of I016a (30.0 g, 149 mmol) in tetrahydrofuran (20 mL) was added dropwise to the reaction solution, and the reaction was continued for 1 hour. DMF (46 mL, 596 mmol) was added dropwise, and the temperature was raised to room temperature for half an hour. The reaction solution was diluted with H2O (100 mL) and extracted with EA (60 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA = 5:1) to obtain compound I016b (9.50 g, 27.8% yield). MS m / z (ESI): 213.0 [M+H] + .

[0474] Step 2:

[0475] 5-Bromophthalazin-1(2H)-one (I016c)

[0476] Compound I016b (9.50 g, 41.5 mmol) was added to a mixture of hydrazine hydrate (7.6 mL, 125 mmol) and acetic acid (50 mL) and allowed to react at 80°C for 16 hours. After completion of the reaction, H₂O (100 mL) was added to the reaction solution. After cooling, a solid precipitated and was filtered. The filter cake was washed with H₂O (15 mL) and dried under reduced pressure without further purification to yield compound I016c (10.2 g). MS m / z (ESI): 227.1 [M+H] + .

[0477] Step 3:

[0478] 5-Bromo-2-methylphthalazin-1(2H)-one (I016d)

[0479] Compound I016c (12.0 g, 53.3 mmol), potassium carbonate (22.0 g, 160 mmol), and iodomethane (30.2 g, 213 mmol) were dissolved in DMF (100 mL) and reacted at 25°C for 4 hours. The reaction solution was extracted with EA (40 mL x 3). The organic phases were combined, washed with H2O (20 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (DCM:CH3OH = 20:1) to obtain compound I016d (12.0 g, yield 94.2%). MS m / z (ESI): 239.1 [M+H] + .

[0480] Step 4:

[0481] 2-Methyl-5-vinylphthalazin-1(2H)-one (I016e)

[0482] Compound I016d (5.20 g, 21.8 mmol), potassium vinyl trifluoroborate (4.38 g, 32.7 mmol), potassium carbonate (9.03 g, 65.4 mmol), and Pd(dppf)Cl2 (0.783 g, 1.07 mmol) were added to a mixture of dioxane (50 mL) and H2O (10 mL) and stirred at 100°C for 3 hours. After completion, the reaction solution was quenched with sodium bicarbonate (30 mL) and extracted with DCM (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The product was then purified by silica gel column chromatography (PE:EA = 5:1) to afford compound I016e (3.00 g, 75.7% yield). MS m / z (ESI): 187.0 [M+H] + .

[0483] Step 5:

[0484] 2-Methyl-1-oxo-1,2-dihydrophthalazine-5-carbaldehyde (I016f)

[0485] Compound I016e (3.00 g, 16.0 mmol) was dissolved in THF (50 mL) and H₂O (200 mL). Sodium periodate (10.3 g, 48.00 mmol) was added to the reaction mixture at 0°C. After stirring for 30 min, potassium (VI) osmate (589 mg, 1.60 mmol) was added, and the reaction mixture was allowed to react at 25°C for 18 hours. The reaction mixture was extracted three times with EA (50 mL). The organic phase was dried, concentrated, and purified by silica gel column chromatography (DCM:CH₃OH = 5:1) to obtain compound I016f (2.50 g, yield 83.1%). MS m / z (ESI): 189.0 [M+H] + .

[0486] Step 6:

[0487] 5-(1-Hydroxypropyl-2-yn-1-yl)-2-methylphthalazin-1(2H)-one (1016g)

[0488] Compound I016f (2.50 g, 13.2 mmol) was dissolved in THF (60 mL). Ethynylmagnesium bromide (53 mL, 26.4 mmol) was added at 0°C under nitrogen protection. The temperature was slowly raised to room temperature and the reaction was allowed to react for 1 hour. After completion, the reaction solution was quenched with saturated ammonium chloride solution (20 mL) and extracted with EA (20 mL x 3). The organic phases were combined, washed with H2O (20 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude compound I016g (2.83 g), which was used directly in the next step without purification. MS m / z (ESI): 215.1 [M+H] + .

[0489] Step 7:

[0490] 1-(2-Methyl-1-oxo-1,2-dihydrophthalazin-5-yl)propyl-2-yn-1-yl acetate (I016h)

[0491] Compound I016g (2.83 g, 13.2 mmol) was dissolved in DCM (30 mL), and triethylamine (4.00 g, 39.6 mmol) and 4-dimethylaminopyridine (2.01 mL, 28.30 mmol) were added. Acetic anhydride (2.47 mL, 26.4 mmol) was added at 0°C, and the mixture was stirred at room temperature for 1 hour. The reaction solution was quenched with H2O (30 mL) and extracted with EA (30 mL x 3). The organic phase was dried, concentrated, and purified by silica gel column chromatography (PE:EA = 10:1) to obtain compound I016h (3.10 g, yield 91.7%). MS m / z (ESI) 257.0 [M+H] + .

[0492] Step 8:

[0493] 5-{1-{[8-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl]amino}propyl-2-yn-1-yl}-2-methylphthalazin-1(2H)-one (I016)

[0494] Compound I016h (2.00 g, 7.81 mmol) and compound I003 (1.38 g, 5.21 mmol) were dissolved in isopropanol (10 mL). Copper(II) trifluoromethanesulfonate (283 mg, 0.781 mmol) and 2,6-bis((4S,5R)-4,5-dihydro-4,5-diphenyloxazol-2-yl)pyridine (408 mg, 0.781 mmol) were then added. Under nitrogen protection and an ice-salt bath, DIPEA (3.37 g, 26.1 mmol) was added to the reaction solution. The reaction solution was slowly warmed to room temperature and stirred for 18 hours. After completion of the reaction, the reaction mixture was concentrated and the crude product was purified by silica gel column chromatography (DCM:CH3OH = 5:1) to afford compound I016 (1.05 g, 41.5% yield). MS m / z (ESI): 462.0 [M+H] + .

[0495] Intermediate 17

[0496] 8-Chloro-N 4 Preparation of -(3,3,3-trifluoro-2,2-dimethylpropyl)pyrido[3,4-d]pyrimidine-4,6-diamine (I017)

[0497] Step 1:

[0498] 3,3,3-Trifluoro-2,2-dimethylpropionamide (I017b)

[0499] Compound I017a (15.6 g, 100 mmol) was dissolved in DCM (150 mL), and CDI (24.3 g, 150 mmol) was added. The mixture was stirred at 25°C for 30 minutes, followed by the addition of 30 mL of aqueous ammonia. The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was separated and the organic phase was washed once with 0.1N HCl solution (70 mL), 0.1N NaOH solution (70 mL), and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford Compound I017b (7.5 g, 48.1% yield) as a white solid. MS m / z (ESI): 156.1 [M+H] + .

[0500] Step 2:

[0501] 3,3,3-Trifluoro-2,2-dimethylpropane-1-amine hydrochloride (I017c)

[0502] Compound I017b (7.5 g, 48.3 mmol) was dissolved in anhydrous THF (100 mL), cooled to 0°C, and under nitrogen, lithium aluminum tetrahydride (3.6 g, 96 mmol) was added portionwise. After complete addition, the mixture was allowed to react at 0°C for 1 hour, then slowly heated to 25°C and reacted for 4 hours. After completion, the reaction was cooled to 0°C and quenched by the dropwise addition of water (5 mL). A 15% sodium hydroxide solution (15 mL) was then added dropwise. The mixture was filtered, and the filtrate was added with a 1N HCl solution (4 mL). The mixture was concentrated to afford Compound I017c (6.8 g, 79% yield) as a white solid.

[0503] Step 3:

[0504] 6-Bromo-8-chloro-N-(3,3,3-trifluoro-2,2-dimethylpropyl)pyrido[3,4-d]pyrimidin-4-amine (I017d)

[0505] Compound I003e (400 mg, 1.53 mmol), compound I017c (814 mg, 4.59 mmol), and PyBOP (1.59 g, 3.06 mmol) were dissolved in anhydrous DMF (5 mL). The temperature was cooled to 0°C. DIPEA (987 mg, 7.65 mmol) was added under nitrogen and allowed to react for 5 hours. After completion of the reaction, the reaction solution was directly purified by reverse-phase column chromatography to obtain compound I017d (140 mg, 23% yield) as a yellow solid. MS m / z (ESI): 384.6 [M+H] + .

[0506] Step 4:

[0507] Tert-butyl {8-chloro-4-[(3,3,3-trifluoro-2,2-dimethylpropyl)amino]pyrido[3,4-d]pyrimidin-6-yl}carbamate (I017e)

[0508] Compound I017d (1.05 g, 2.73 mmol), tert-butyl carbamate (351 mg, 3.00 mmol), Xantphos Pd G2 (243 mg, 0.273 mmol), and cesium carbonate (1.78 g, 8.19 mmol) were dissolved in 1,4-dioxane (11 mL) and stirred at 90°C under nitrogen for 16 hours. After completion, the reaction was cooled to room temperature and quenched with water (50 mL). The mixture was extracted with EA (50 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1) to obtain Compound I017e (1.00 g, 87.26% yield) as a pale yellow solid. MS m / z (ESI): 420.1 [M+H] + .

[0509] Step 5:

[0510] 8-Chloro-N 4 -(3,3,3-trifluoro-2,2-dimethylpropyl)pyrido[3,4-d]pyrimidine-4,6-diamine (I017)

[0511] Compound I017e (1.00 g, 2.38 mmol) was added to a 1,4-dioxane / HCl (4N, 10 mL) solution and reacted at room temperature for 6 hours. The mixture was then concentrated under reduced pressure and diluted with DCM (10 mL) and H2O (10 mL). The pH was adjusted to approximately 7 with saturated sodium bicarbonate solution. Extraction was performed with DCM (20 mL x 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE:EA = 1:1) to obtain Compound I017 (600 mg, yield 78.85%) as a yellow solid. MS m / z (ESI): 320.1 [M+H] + .

[0512] Preparation Example

[0513] Example 1

[0514] Preparation of (S)-5-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl][(8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl)amino]methyl}-2-methylisoquinolin-1-(2H)-one (E001)

[0515] To a solution of compound I003 (200 mg, 0.6 mmol) and compound I002 (97 mg, 0.3 mmol) in 1,4-dioxane (10 mL) were added XantPhos Pd-G2 (53 mg, 0.06 mmol) and sodium tert-butoxide (115 mg, 1.2 mmol). The mixture was heated to 120°C under nitrogen for 3 hours. After completion, the reaction was cooled to room temperature, filtered, and concentrated to obtain the crude product, which was then purified by prep-HPLC (Waters 2767, SunFire Prep C18 OBD™, 19x250 mm and 10 μm columns; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 20-80%; retention time: 5.3-10.5 min, 16 min total) to afford compound E001 (15 mg, 4.4% yield) as a yellow solid. MS m / z(ESI):570.3[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.24(s,1H),8.19(d,J=7.9Hz,1H),8.02(d,J=7.3Hz,3H),7.77(d,J=7.4Hz,1H),7.53-7.44(m,2H),7.2 0(s,1H),6.82(dd,J=17.7,8.0Hz,2H),3.53(s,1H),3.49(s,3H),3.37(d,J=5.8Hz,1H),2.67(s,1H),2.30(s,6H),0.90(s,9H).

[0516] Example 2

[0517] Preparation of (S)-6-{[(1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl)(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)methyl]amino}-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (E002)

[0518] Compound E001 (60 mg, 0.1 mmol), zinc cyanide (35 mg, 0.3 mmol), and XantPhos Pd-G2 (9 mg, 0.01 mmol) were dissolved in anhydrous 1,4-dioxane (10 mL). The reaction was heated to 120°C under nitrogen for 3 hours. After completion of the reaction, the mixture was cooled to room temperature, filtered, and concentrated to obtain the crude product, which was then purified by prep-HPLC (Waters 2767, column: SunFire Prep C18 OBD™, 19*250 mm, 10 μm; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 48-58%; retention time: 9.2-10.0 min, total 16 min) to afford compound E002 (9 mg, 16% yield) as a yellow solid. MS m / z (ESI): 561.3 [M+H] + . 1 H NMR (400MHz, MeOD) δ8.31(d,J=8.0Hz,1H),8.23(s,1H),7.84(s,1H),7.72(d,J=7.4Hz,1H),7.48(t,J=7.8Hz,1H),7.39(d,J=7.7H z,1H),7.29(s,1H),6.97(s,1H),6.90(d,J=7.7Hz,1H),3.60(s,3H),3.51(d,J=4.6Hz,2H),2.67(s,1H),2.37(s,6H),0.96(s,9H).

[0519] Example 3

[0520] (S)-N 6 -{{1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methyl}-8-chloro-N 4 Preparation of neopentylpyrido[3,4-d]pyrimidine-4,6-diamine (E003)

[0521] Compound I005 (120 mg, 0.418 mmol), compound I003 (275 mg, 0.835 mmol), XantPhos PdG2 (74.2 mg, 0.0835 mmol), and sodium tert-butoxide (160 mg, 1.67 mmol) were added to dioxane (3 mL) at room temperature and heated to 120°C under nitrogen for 2 hours. After completion, the reaction was cooled to room temperature, concentrated under reduced pressure, and purified by preparative HPLC (Waters 2767 / QDA, column: SunFire C18, 19x250 mm, 10 μm; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 46-49%; retention time: 8.3-11.2 min, 20 min total) to afford compound E003 (8.98 mg, 4.01% yield). MS m / z(ESI):536.2[M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.24(s,1H),8.19(s,1H),8.06-7.96(m,3H),7.14( s,1H),6.97(dd,J=8.3,3.0Hz,1H),6.40(d,J=8.2Hz,1H),3.49(d,J=6.6Hz,1 H),3.40(d,J=6.0Hz,1H),2.60(s,3H),1.55-1.49(m,1H),1.16(t,J=6.0Hz,2 H),0.96(d,J=1.6Hz,2H),0.92(s,9H),0.50-0.44(m,2H),0.31-0.27(m,2H).

[0522] Example 4

[0523] Preparation of (S)-6-{{{1-{[1,1'-bis(cyclopropane)]-1-yl}-1H-1,2,3-triazol-4-yl}(6-fluoro-2-methylpyridin-3-yl)methyl}amino}-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (E004)

[0524] To a solution of E003 (60 mg, 0.112 mmol) in dioxane (10 mL) at room temperature were added XantPhos Pd-G2 (9.96 mg, 0.0112 mmol) and zinc cyanide (26.2 mg, 0.224 mmol). The mixture was then heated to 120°C and reacted under nitrogen for 16 hours. After completion, the reaction was cooled to room temperature and concentrated under reduced pressure. The mixture was purified by preparative HPLC (Waters 2767 / Qda, column: Pursuit XRs 10 C18 250*21.2 mm, 10 μm; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 63-73%; retention time: 8.4-9.2 min, 16 min total) to afford compound E004 (10.05 mg, 17.05% yield). MS m / z(ESI):527.2[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.39(t,J=6.2Hz,1H),8.29(s,1H),8.22(d,J=7.8Hz,1 H),8.19(s,1H),8.00(t,J=8.3Hz,1H),7.56(s,1H),6.98(dd,J=8.4,2.9Hz,1H ),6.45(d,J=7.8Hz,1H),3.54-3.43(m,2H),2.59(s,3H),1.56-1.44(m,1H),1. 16(q,J=5.5Hz,2H),0.99-0.89(m,11H),0.55-0.42(m,2H),0.33-0.24(m,2H).

[0525] Example 5

[0526] (S)-N6-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methyl}-8-chloro-N 4 Preparation of neopentylpyridinyl[3,4-d]pyrimidine-4,6-diamine (E005)

[0527] To a solution of I006 (80 mg, 0.293 mmol) and I003 (193 mg, 0.585 mmol) in dioxane (10 mL) were added XantPhos Pd-G2 (52.0 mg, 0.0585 mmol) and sodium tert-butoxide (112 mg, 1.17 mmol) at room temperature. The mixture was then heated to 120°C and reacted under nitrogen for 2 hours. After completion, the reaction was cooled to room temperature and concentrated under reduced pressure. Purification was performed by preparative HPLC (Waters 2767 / QDA, column: SunFire C18, 19x250 mm, 10 μm; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 47-47%; retention time: 7.8-10.7 min, 16 min total) to afford compound E005 (3.59 mg, 2.35% yield). MS m / z(ESI):522.2[M+H] + . 1 H NMR(400MHz,MeOD)δ8.22(s,1H),7.97-7.88(m,2H),6.92(s,1H),6.87(dd,J=8.4,2.4 Hz,1H),6.53(s,1H),3.51(s,2H),2.69(s,1H),2.63(s,3H),2.39(s,6H),0.98(s,9H).

[0528] Example 6

[0529] Preparation of (S)-6-{{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl](6-fluoro-2-methylpyridin-3-yl)methyl}amino}-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (E006)

[0530] To a solution of I006 (60 mg, 0.220 mmol) and I010 (121 mg, 0.439 mmol) in dioxane (10 mL) were added BrettPhos Pd-G4 (40.4 mg, 0.0439 mmol) and sodium tert-butoxide (84.3 mg, 0.878 mmol) at room temperature, and the temperature was raised to 120°C and the reaction was carried out under nitrogen for 2 hours. After the reaction was complete, the temperature was lowered, the mixture was concentrated under reduced pressure, and the mixture was purified by preparative HPLC (Waters 2767 / QDA, column type: Pursuit XRs 10 C18 250*21.2 mm, 10 μm; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 62-68%; retention time: 7.6-10 min, total 16 min) to obtain compound E006 (11.03 mg, yield 9.80%). MS m / z (ESI): 513.3 [M+H] + . 1 H NMR(400MHz,DMSO-d6) δ8.40(t,J=6.4Hz,1H),8.28(s,1H),8.23(d,J=7.8Hz,1H),8.09(s,1H),7.97(t,J=8.3Hz,1H),7.55(s,1H),6.98(dd,J=8.5,3.0Hz,1H) ,6.48(d,J=7.8Hz,1H),3.50(dd,J=13.0,6.4Hz,1H),3.39(dd,J=13.0,6.1Hz,1H),2.69(s,1H),2.58(s,3H),2.32(s,6H),0.93(s,9H).

[0531] Example 7

[0532] Preparation of (S)-5-{{[8-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl]amino}{1-[1-(difluoromethyl)cyclopropyl]-1H-1,2,3-triazol-4-yl}methyl}-2-methylisoquinolin-1-(2H)-one (E007)

[0533] Compound I007 (30 mg, 0.087 mmol) and compound I003 (42 mg, 0.13 mmol) were added to dioxane (10 mL), followed by Xantphos-Pd-G2 (7 mg, 0.008 mmol) and sodium tert-butoxide (16 mg, 0.174 mmol). The mixture was reacted at 120°C under nitrogen for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by preparative HPLC (Waters 2767 / Qda, column: XBridge XBridge C18, 19*250 mm and 10 μm; mobile phase A: 0.05% NH3·H2O / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 52%-59%; retention time: 8.2-8.7 min, 16 min total) to afford compound E007 (12.19 mg, 23.5% yield). MS m / z(ESI):594.1[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.23(s,1H),8.19(d,J=9.0Hz,2H),8.03-7.94(m,2H),7.76(d,J=7.3Hz,1H),7.53-7.44(m,2H),7.15(s,1H),6.87 (d,J=7.7Hz,1H),6.80(d,J=8.3Hz,1H),6.11(t,J=54.1Hz,1H),3.49(s,3H),3.46(s,1H),3.38(d,J=5.9Hz,1H),1.49(s,4H),0.90(s,9H).

[0534] Example 8

[0535] Preparation of (S)-6-{{{1-[1-(difluoromethyl)cyclopropyl]-1H-1,2,3-triazol-4-yl}(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)methyl}amino}-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (E008)

[0536] Compound E007 (60 mg, 0.101 mmol) and zinc cyanide (47.3 mg, 0.404 mmol) were dissolved in dioxane (1.5 mL), and XantPhos Pd G2 (13.5 mg, 0.0152 mmol) was added. The reaction liquid was protected with nitrogen and stirred at 120°C for 4 hours. After completion of the reaction, the product was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 0-95%) and then by preparative HPLC (Waters 2767 / Qda, column type: Pursuit XRs 10 C18, 250*21.2 mm, 10 μm; mobile phase A: 0.1% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 49%-54%; retention time: 9.1-10.3 min, total 16 min) to obtain solid compound E008 (10.79 mg, yield 18.3%). MS m / z (ESI): 585.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.37(t,J=6.0Hz,1H),8.29(d,J=10.3Hz,2H),8.23-8.12(m,2H),7.77(dd,J=7.5,0.9Hz,1H),7.58(s,1H),7.5 4-7.41(m,2H),6.86(t,J=9.0Hz,2H),6.10(m,1H),3.50(s,3H),3.47(d,J=6.3Hz,1H),3.41(d,J=6.2Hz,1H),1.48(s,4H),0.92(s,9H).

[0537] Example 9

[0538] Preparation of 5-{(S)-[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]{{8-chloro-4-{[(R)-3,3-dimethyltetrahydro-2H-pyran-4-yl]amino}pyridin[3,4-d]pyrimidin-6-yl}aminomethyl}}-2-methylisoquinolin-1-(2H)-one (E009)

[0539] At room temperature, compound I008 (100 mg, 0.27 mmol) and compound I002 (86 mg, 0.27 mmol) were dissolved in 1,4-dioxane (2.5 mL), and XantPhos Pd G2 (24 mg, 0.027 mmol) and sodium tert-butoxide (52 mg, 0.52 mmol) were added. The mixture was stirred at 120°C under nitrogen for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. The crude product was first purified by silica gel column chromatography (DCM:MeOH = 50:1) to obtain a yellow solid racemate (70 mg, 42.4% yield). Compound E009 (18.39 mg) was then separated by SFC. MS m / z (ESI): 612.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.25(s,1H),8.17(d,J=8.1Hz,1H),8.08-8.03(m,2H),7.82(d,J=7.4Hz,1H),7.51( d,J=7.7Hz,1H),7.46(t,J=7.7Hz,1H),7.39(d,J=9.4Hz,1H),7.12(s,1H),6.91(d,J=7.6Hz,1H),6.82(d,J= 8.7Hz, 1H), 4.47 (d, J = 8.4Hz, 1H), 3.93 (d, J = 7.8Hz, 1H), 3.49 (s, 3H), 3.43 (d, J = 11.3Hz, 2H), 3.17 (d, J = 11. 2Hz,1H),2.68(s,1H),2.31(s,6H),1.88(d,J=8.0Hz,1H),1.55(d,J=12.0Hz,1H),0.96(s,3H),0.64(s,3H).

[0540] Example 10

[0541] Preparation of 5-{(1S)-[(8-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl)amino][1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylisoquinolin-1(2H)-one (E010)

[0542] Compound I003 (36.92 mg, 0.11 mmol) and compound I009 (30 mg, 0.09 mmol) were dissolved in 1,4-dioxane (2 mL), and XantPhos Pd G2 (8.27 mg, 0.009 mmol) and t-BuONa (26.81 mg, 0.28 mmol) were added. The mixture was stirred at 120°C under nitrogen for 2 hours. After completion, the reaction solution was filtered, concentrated, and purified by preparative HPLC (Waters 2767 / Qda, column: XBridge C18 19*250 mm and 10 μm; mobile phase A: 0.05% FA / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 40%-52%; retention time: 7.3-9.2 min, total 16 min) to afford compound E010 (6.78 mg, 12.78% yield) as a yellow solid. MS m / z(ESI):570.1[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.23 (s, 1H), 8.18 (d, J = 7.3Hz, 1H), 7.97 (s, 2H), 7.76 (s, 1H), 7.54-7.41 (m, 2H), 7.16(s,1H),6.84(s,1H),6.74(d,J=7.4Hz,1H),4.21(s,1H),3.49(s,5H),1.64(s,2H),1.23(s,1H),1.07(s,1H),0.86(m,11H).

[0543] Example 11

[0544] Preparation of 6-{{(1S)-(2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)[1-(spiro[2.2]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}amino}-4-(neopentylamino)pyrido[3,4-d]pyrimidine-8-carbonitrile (E011)

[0545] Compound E010 (40 mg, 0.07 mmol) and zinc cyanide (41 mg, 0.35 mmol) were added to dioxane (2.5 mL), followed by Xantphos-Pd-G2 (6 mg, 0.007 mmol). The mixture was reacted at 120°C under nitrogen for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by preparative HPLC (Waters 2767 / Qda, column: XBridge C18 19*250 mm and 10 μm; mobile phase A: 0.05% NH3H2O / H2O, mobile phase B: MeCN; flow rate: 20 mL / min; gradient: 54%-61%; retention time: 8.5-8.9 min, total 16 min) to afford compound E011 (7.84 mg, 20.1% yield) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.34(s,1H),8.28(s,1H),8.24-8.18(m,2H),7.96(s,1H),7.76(t,J=6.5Hz,1H),7.57(s,1H),7.51-7.44(m,2H),6.81(d, J=7.5Hz,2H),4.22(s,1H),3.50(s,4H),3.41-3.36(m,1H),1.64(s,2H), 1.09(d,J=9.2Hz,1H),0.96(s,2H),0.92(s,9H),0.80(d,J=8.3Hz,1H).MS m / z(ESI):561.3[M+H] + .

[0546] Example 12

[0547] Preparation of (S)-5-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]{{8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl}amino}methyl}-8-fluoro-2-methylisoquinolin-1(2H)-one (E012)

[0548] Step 1:

[0549] 5-{1-{[8-chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl]amino}prop-2-yn-1-yl}-8-fluoro-2-methylisoquinolin-1(2H)-one (E012a)

[0550] Compound I011 (700 mg, 2.64 mmol), compound I012 (1.1 g, 4.03 mmol), and Cu(OTf)2 (190 mg, 0.53 mmol) were dissolved in isopropanol (10 mL). DIPEA (1.7 g, 13.2 mmol) and 2,6-bis((4R,5S)-4,5-diphenyl-4,5-dihydrooxazol-2-yl)pyridine (276 mg, 0.53 mmol) were added to the reaction solution. The atmosphere was replaced with nitrogen and the reaction was continued at 25°C for 18 hours. After completion of the reaction, the reaction solution was concentrated and purified by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain Compound E012a (950 mg, 75% yield) as a yellow solid.

[0551] Step 2:

[0552] (S)-5-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]{{8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl}amino}methyl}-8-fluoro-2-methylisoquinolin-1(2H)-one (E012)

[0553] To a solution of compound E012a (550 mg, 1.15 mmol) in 2-methyltetrahydrofuran (10 mL) at room temperature were added copper sulfate pentahydrate (167 mg, 0.67 mmol) and sodium ascorbate (132 mg, 0.67 mmol). Then, a solution of 1-azido-bicyclo[1.1.1]pentane in acetonitrile (100 mg / mL, 1.7 mL, 1.72 mmol) was added, and the reaction was stirred at 75°C under nitrogen for 16 hours. After the reaction, the product was concentrated under reduced pressure and purified by silica gel column chromatography (PE:EA = 100%) to obtain a crude product. The crude product was then separated by SFC (column type: Daicel ChiralPak AD, 40 mm ID x 250 mm, 10 μm; mobile phase A: Supercritical CO2, mobile phase B: MeOH, A:B = 0:50; flow rate: 60 mL / min; column temperature: 38°C; wavelength: 210 nm) to obtain compound E012 (239 mg, 35% yield) as a yellow solid. MS m / z (ESI): 588.4 [M+H] + . 1H NMR (400MHz, MeOD-d4) δ8.21 (s, 1H), 7.84 (s, 1H), 7.68-7.65 (m, 1H), 7.43 (d, J = 7.6Hz, 1H), 7.15-7. 11(m,1H),6.88-6.79(m,3H),3.55(s,3H),3.52-3.45(m,2H),2.67(s,1H),2.36(s,6H),0.94(s,9H).

[0554] Example 13

[0555] Preparation of (S)-5-{{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl][8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl]amino}methyl}-1-methylquinolin-2(1H)-one (E013)

[0556] To a solution of compound I013 (70 mg, 0.2 mmol) and I003 (130 mg, 0.4 mmol) in 1,4-dioxane (10 mL) at room temperature were added XantPhos Pd-G2 (55 mg, 0.06 mmol) and sodium tert-butoxide (85 mg, 1.0 mmol). The reaction was then heated to 70°C and allowed to react under nitrogen for 2 hours. After completion of the reaction, the temperature was lowered, the mixture was filtered, and the filtrate was concentrated and purified by prep-HPLC (Waters 2767 / QDA, column: SunFire C18, 19*250 mm, 10 μm; mobile phase A: 0.05% NH3H2O / H2O, mobile phase B: ACN; flow rate: 20 mL / min; gradient: 49-49%; retention time: 6.7-8.1 min; total 16 min) to afford E013 (11.99 mg, 5.4% yield) as a yellow solid. MS m / z(ESI):570.3[M+H] + . 1 H NMR (400MHz, MeOD-d4) δ8.31(d,J=9.8Hz,1H),8.21(s,1H),7.84(s,1H),7.65-7.54(m,2H),7.39(d,J=7.2Hz,1H),6.96(s,1H),6.8 7(s,1H),6.71(d,J=9.8Hz,1H),3.76(s,3H),3.52(d,J=13.4Hz,1H),3.44(d,J=13.1Hz,1H),2.67(s,1H),2.36(s,6H),0.94(s,9H).

[0557] Example 14

[0558] (S)-N 6 -{[1,2,4]triazolo[3,4-a]isoquinolin-7-yl[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-8-chloro-N 4 Preparation of neopentylpyrido[3,4-d]pyrimidine-4,6-diamine (E014)

[0559] Step 1:

[0560] N 6 -(1-([1,2,4]triazolo[3,4-a]isoquinolin-7-yl)prop-2-yn-1-yl)-8-chloro-N 4 -Neopentylpyrido[3,4-d]pyrimidine-4,6-diamine (E014a)

[0561] Compound I011 (700 mg, 2.64 mmol), I014 (700 mg, 2.64 mmol), and Cu(OTf)2 (190 mg, 0.53 mmol) were dissolved in isopropanol (10 mL). DIPEA (1.7 g, 13.2 mmol) and 2,6-bis((4R,5S)-4,5-diphenyl-4,5-dihydrooxazol-2-yl)pyridine (276 mg, 0.53 mmol) were added to the reaction solution. The atmosphere was replaced with nitrogen and the reaction was continued at 25°C for 18 hours. After completion, the reaction solution was concentrated and purified by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound E014a (300 mg, 24% yield) as a yellow solid.

[0562] Step 2:

[0563] (S)-N 6 -{[1,2,4]triazolo[3,4-a]isoquinolin-7-yl[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-8-chloro-N 4 -Neopentylpyrido[3,4-d]pyrimidine-4,6-diamine (E014)

[0564] To a solution of compound E014a (300 mg, 0.628 mmol) in 2-methyltetrahydrofuran (10 mL) at room temperature were added copper sulfate pentahydrate (80 mg, 0.32 mmol) and sodium ascorbate (63.4 mg, 0.32 mmol). A solution of 1-azido-bicyclo[1.1.1]pentane in acetonitrile (100 mg / mL, 1 mL, 0.94 mmol) was then added, and the mixture was stirred at 75°C under nitrogen for 16 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA = 100%) to obtain the crude product. The crude product was then subjected to chiral separation to afford compound E014 (118 mg, 40% yield) as a yellow solid. MS m / z (ESI): 580.2 [M+H] + . 1 H NMR(400MHz,MeOD-d4)δ9.07(s,1H),8.48(d,J=7.9Hz,1H),8.18(d,J=7.6Hz,1H),8.11(s,1H),7.80(s,1H),7.72(d,J=7.5Hz,1H),7.62(t,J =7.8Hz,1H),7.53(d,J=7.6Hz,1H),6.96(s,1H),6.82(s,1H),3.38(q,J=13.3Hz,2H),3.21(dt,J=3.2,1.6Hz,6H),2.57(s,1H),0.82(s,9H).

[0565] Example 15

[0566] Preparation of (S)-8-{{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl][8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl]amino}methyl}-2H-benzo[b][1,4]oxazin-3(4H)-one (E015)

[0567] Step 1:

[0568] 8-{{1-[8-Chloro-4-(neopentylamino)pyrido[3,4-d]pyrimidin-6-yl]amino}prop-2-yn-1-yl}-2H-benzo[b][1,4]oxazin-3(4H)-one (E015a)

[0569] At room temperature, compound I015 (1.0 g, 4.0 mmol) and I011 (700.0 mg, 2.7 mmol) were dissolved in isopropanol (20 mL). The ligand 2,6-bis((4R,5S)-4,5-diphenyl-4,5-dihydrooxazol-2-yl)pyridine (210.0 mg, 0.4 mmol) and copper trifluoromethanesulfonate (145.0 mg, 0.4 mmol) were added. The temperature was cooled to 0°C under nitrogen, and DIPEA (1.4 g, 10.8 mmol) was added dropwise. The reaction was stirred at 0°C for 10 hours. After completion of the reaction, the reaction solution was concentrated and purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound E015a (550.0 mg, 45.8% yield) as a yellow solid. MS m / z (ESI): 451.2 [M+H] + .

[0570] Step 2:

[0571] (S)-8-{{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl][8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl]amino}methyl}-2H-benzo[b][1,4]oxazin-3(4H)-one (E015)

[0572] Compound E015a (550.0 mg, 1.2 mmol), copper sulfate pentahydrate (150.0 mg, 0.6 mmol), and sodium ascorbate (120.0 mg, 0.6 mmol) were dissolved in 2-methyltetrahydrofuran (20 mL), and 1-azidobicyclo[1.1.1]pentane (1.8 mL, 1.8 mmol, 1 M) was added. The reaction mixture was stirred at 75°C overnight under nitrogen. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE:EA = 1:2) to obtain the crude product. The crude product was then purified by prep-HPLC (A: 0.05% NH3H2O / H2O, B: ACN; flow rate: 20 mL / min; gradient: 45-50%; retention time: 6.5-7.5 min; total 16 min) to obtain compound E015 (195.7 mg, 29.12% yield). MS m / z(ESI):560.3[M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.72(s,1H),8.24(s,1H),8.07(t,J=6.2Hz,1H),7.9 8(s,1H),7.67(d,J=8.9Hz,1H),7.14(t,J=3.1Hz,2H),6.92(t,J=7.8Hz,1H), 6.82(dd,J=7.8,1.3Hz,1H),6.54(d,J=8.8Hz,1H),4.59(s,2H),3.55(dd,J=1 3.0, 6.6Hz, 1H), 3.32 (d, J = 5.9Hz, 1H), 2.68 (s, 1H), 2.31 (s, 6H), 0.92 (s, 9H).

[0573] Example 16

[0574] Preparation of (S)-5-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]{[8-chloro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl]amino}methyl}-2-methylphthalic acid-1(2H)-one (E016)

[0575] To a solution of compound I016 (1.05 g, 2.27 mmol) in 2-methyltetrahydrofuran (10 mL) at room temperature were added copper sulfate pentahydrate (285 mg, 1.14 mmol) and sodium ascorbate (226 mg, 1.14 mmol). A solution of 1-azido-bicyclo[1.1.1]pentane in acetonitrile (100 mg / mL, 5 mL, 4.54 mmol) was then added, and the mixture was stirred at 75°C under nitrogen for 16 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (EA / PE = 100%) to obtain the crude product, which was then purified by preparative HPLC (Gilson GX-281 column: XSelect XHS C18, 19*250 mm and 10 μm; mobile phase A: 10 mmol NH 3` Yellow solid compound E016 (498 mg, 37.8% yield) was obtained by purification and separation using H2O, mobile phase B: ACN; flow rate: 20 mL / min; gradient: 49% to 54%; retention time: 9-10 min, total 18 min. MS m / z (ESI): 572.1 [M+H] + . 1H NMR(400MHz,MeOD-d6)δ8.70(s,1H),8.32(d,J=8.0Hz,1H),8.23(s,1H),7.98-7.93(m,2H),7.80(t,J=7.6 Hz,1H),7.05(s,1H),6.94(s,1H),3.82(s,3H),3.56-3.46(m,2H),2.68(s,1H),2.38(s,6H),0.95(s,9H).

[0576] Example 17

[0577] Preparation of (S)-5-{{1-[bicyclo[1.1.1]pentan-1-yl]-1H-1,2,3-triazol-4-yl}{[8-fluoro-4-(neopentylamino)pyridin[3,4-d]pyrimidin-6-yl]amino}methyl}-2-methylisoquinolin-1-(2H)-one (E017)

[0578] At room temperature, compound E001 (100 mg, 0.175 mmol) and CsF (264 mg, 1.75 mmol) were dissolved in NMP (1.5 mL). The reaction mixture was stirred at 165°C under nitrogen for 1 hour. After completion of the reaction, the mixture was lyophilized and purified by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain the racemic form of compound E017 (21 mg, 42.4% yield) as a yellow solid. Compound E017 (13.32 mg) was then resolved by SFC (15-25% MeOH DEA C2 AS). MS m / z (ESI): 554.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=8.0Hz,2H),8.00-7.90(m,3H),7.75(d,J=7.4Hz,1H),7.52-7.44(m,2H),7.05(s,1H),6.80 (d,J=7.8Hz,1H),6.74(d,J=8.5Hz,1H),3.48(d,J=8.1Hz,4H),3.37(d,J=5.8Hz,1H),2.67(s,1H),2.30(s,6H),0.91(s,9H).

[0579] Example 18

[0580] Preparation of (S)-5-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]{{8-chloro-4-[(3,3,3-trifluoro-2,2-dimethylpropyl)amino]pyridin[3,4-d]pyrimidin-6-yl}amino}methyl}-2-methylphthalazin-1(2H)-one (E018)

[0581] Step 1:

[0582] 5-{1-{{8-chloro-4-[(3,3,3-trifluoro-2,2-dimethylpropyl)amino]pyrido[3,4-d]pyrimidin-6-yl}amino}propyl-2-yn-1-yl}-2-methylphthalazin-1(2H)-one (E018a)

[0583] Compound I017 (320 mg, 1.00 mmol) and compound I016h (384 mg, 1.50 mmol) were dissolved in isopropanol (5 mL), followed by the addition of copper(II) trifluoromethanesulfonate (54.3 mg, 0.150 mmol) and 2,6-bis((4R,5S)-4,5-diphenyl-4,5-dihydrooxazol-2-yl)pyridine (78.3 mg, 0.150 mmol). Under nitrogen protection and an ice-salt bath, DIPEA (645 mg, 5.00 mmol) was added to the reaction solution. The reaction solution was slowly warmed to room temperature and stirred for 18 hours. After completion of the reaction, the solution was concentrated and the crude product was purified by silica gel column chromatography (DCM:CH3OH = 5:1) to afford compound E018a (220 mg, 42.64% yield) as a yellow solid. MS m / z (ESI): 516.0 [M+H] + .

[0584] Step 2:

[0585] (S)-5-{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]{{8-chloro-4-[(3,3,3-trifluoro-2,2-dimethylpropyl)amino]pyridin[3,4-d]pyrimidin-6-yl}amino}methyl}-2-methylphthalazin-1(2H)-one (E018)

[0586] To a solution of compound E018a (220 mg, 0.426 mmol) in 2-methyltetrahydrofuran (10 mL) at room temperature were added copper sulfate pentahydrate (53.3 mg, 0.213 mmol) and sodium ascorbate (42.2 mg, 0.213 mmol). Then, a solution of 1-azido-bicyclo[1.1.1]pentane in acetonitrile (100 mg / mL, 1 mL, 0.852 mmol) was added, and the reaction was stirred at 75°C under nitrogen for 16 hours. After completion of the reaction, the product was concentrated under reduced pressure and purified by silica gel column chromatography (EA / PE = 100%) to obtain the crude product. This was then purified and isolated by preparative HPLC (Waters 2767 / Qda column: XBridge XBridge C18, 19*250 mm and 10 μm; mobile phase A: 0.03% mmol NH3H2O / H2O, mobile phase B: ACN; flow rate: 20 mL / min; gradient: 45% to 49%; retention time: 8.5-10.4 min, 16 min total) to obtain compound E018 (34.4 mg, 12.9% yield) as a yellow solid. MS m / z (ESI): 625.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.73(s,1H),8.32-8.24(m,2H),8.21(d,J=8.1Hz,1H),8.13(s,2H),8.00(d,J=7.7Hz,1H),7.82(t,J=7.7Hz,1H),7.12 (s,1H),6.92(d,J=8.0Hz,1H),3.90(dd,J=13.7,6.7Hz,1H),3.72(s,3H),3.70-3.65(m,1H),2.69(s,1H),2.31(s,6H),1.10(d,J=3.6Hz,6H).

[0587] Example 19

[0588] Preparation of (S)-5-{{{8-chloro-4-[(3,3,3-trifluoro-2,2-dimethylpropyl)amino]pyrido[3,4-d]pyrimidin-6-yl}amino}[1-(3-methylbicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylphthalazin-1(2H)-one (E019)

[0589] Step 1:

[0590] 1-Azido-3-methylbicyclo[1.1.1]pentane (E019b)

[0591] Compound E019a (100 mg, 0.75 mmol) and DIPEA (192 mg, 1.5 mmol) were dissolved in acetonitrile (5 mL). AMDP (300 mg, 1.04 mmol) was added portionwise and stirred at room temperature for 16 hours. After the reaction, the mixture was used directly in the next step without further treatment.

[0592] Step 2:

[0593] (S)-5-{{{8-Chloro-4-[(3,3,3-trifluoro-2,2-dimethylpropyl)amino]pyrido[3,4-d]pyrimidin-6-yl}amino}[1-(3-methylbicyclo[1.1.1]pentan-1-yl)-1H-1,2,3-triazol-4-yl]methyl}-2-methylphthalazin-1(2H)-one (E019)

[0594] Compound I016 (200 mg, 0.38 mmol) and E019b (3.2 mL, 0.48 mmol, 0.15 mmol / mL) were dissolved in 2-methyltetrahydrofuran (5 mL), and copper sulfate pentahydrate (48 mg, 0.19 mmol) and sodium L-ascorbate (38 mg, 0.19 mmol) were added. The reaction mixture was stirred at 75°C under nitrogen for 16 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM:CH3OH = 0-10%) to obtain the racemic form of solid compound E012 (80 mg, 33.05% yield). Compound E019 (57.82 mg) was then resolved by SFC (15% MEOH DEA C4 AS). MS m / z (ESI): 639.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.73(s,1H),8.33-8.18(m,4H),8.11(s,1H),8.00(d,J=7.3Hz,1H),7.81(t,J=7.8Hz,1H),7.13(s,1H) ,6.93(d,J=7.9Hz,1H),3.89(d,J=6.8Hz,1H),3.72(s,3H),3.70-3.65(m,1H),2.18(s,6H),1.31(s,3H),1.10(d,J=3.6Hz,6H).

[0595] The following compounds were prepared according to the examples and procedures described herein using appropriate starting materials and appropriate protecting group chemistries as required.

[0596] Comparative Example 1

[0597] The structure of compound (S)-5-{{[1-(bicyclo[1.1.1]pentan-1-yl)-1H-imidazol-4-yl][(8-chloro-4-(neopentylamino)quinazolin-6-yl)]amino}methyl}-2-methylisoquinolin-1(2H)-one is:

[0598] The synthetic route of the compound of Comparative Example 1 refers to the synthetic route of the compound numbered I-27 in the specification of International Patent No. WO2018 / 005435.

[0599] Biological test data

[0600] Test Example 1: High-throughput Cot biochemical assay

[0601] Human Cot enzyme activity was measured using PhosphoSens (AssayQuant), based on the chelation enhanced fluorescence (ChEF) principle. In this assay, Cot catalyzes the phosphorylation of a peptide substrate containing an 8-hydroxyquinoline derivative side chain. After the peptide substrate is phosphorylated, the phosphate group is chelated with the 8-hydroxyquinoline side chain near the phosphorylation site by a magnesium ion, and the chelated state can be detected by fluorescence. Purified recombinant human Cot catalytic domain (30-397 amino acids) was purchased from Carna Biosciences. Compounds dissolved in DMSO were delivered to Proxi white low-capacity 384-well plates at the desired concentration using an Echo 650 acoustic liquid dispenser. 5 μL of Cot enzyme dissolved in reaction buffer was transferred to the assay plate. The reaction buffer was 50 mM HEPES, pH 7.5, 0.01% Brij-35, 10 mM MgCl2, 1.2 mM DTT, 0.5 mM EGTA, 50 μM ATP, and 0.2 mg / mL BSA. After incubation at room temperature for 15 minutes, 5 μL of the peptide substrate dissolved in the reaction buffer was added. The Ensight reader (PerkinElmer) was then used with excitation at 485 nm and emission at 535 nm to read the values. The percentage of inhibition was calculated as follows:

[0602] % inhibition = (maximum signal - compound signal) / (maximum signal - minimum signal) × 100

[0603] “+++” represents IC 50 <10nM, “++” represents IC 50 ≥10nM and IC 50 ≤100nM, “+” represents IC50 >100nM.

[0604] Test Example 2: Cot monocyte TNFα cell-based assay

[0605] Cryopreserved human monocytes (TPCS) were thawed and diluted to 0.125×10 in RPMI medium containing Glutamax (10 mM HEPES, 1× Pen-Strep, 55 μM β-mercaptoethanol, 1 mM sodium pyruvate) and 10% FBS. 6 cells / mL and recovered at 37°C for 2 hours. The cell suspension was then plated on a black 384-well clear bottom plate (Corning) at a density of 5000 cells / well. The plate was pre-spotted with the test compound and serially diluted in DMSO with a final DMSO concentration of 0.1%. The plated cells were treated with the compound at 37°C for 1 hour. The cells were then stimulated with 50pg / mL of LPS (Sigma), excluding the outer row of the plate for unstimulated cell control wells. The cells were incubated at 37°C for another 4 hours, and then the cells were centrifuged to take 15μL of cell supernatant and analyzed for TNFα content using an ELISA human TNFα detection system (R&D).

[0606] “+++” represents IC 50 <10nM, “++” represents IC 50 ≥10nM and IC 50 ≤100nM, “+” represents IC 50 >100nM, “ND” means not measured.

[0607] As shown in Table 1, the compounds of Formula I are inhibitors of Cot (Osaka thyroid carcinoma) kinase.

[0608] Table 1

[0609] The experimental results in Table 1 above show that the compounds disclosed in the present invention are effective inhibitors of Osaka thyroid carcinoma (Cot) kinase and have significant inhibitory activity on TNFa production, which suggests that the compounds of the present invention can be used to treat Cot-related diseases such as inflammation.

[0610] Test Example 3: Pharmacokinetic Evaluation in Mice

[0611] The in vivo pharmacokinetic properties of the compounds were tested using CD-1 (ICR) mice (male, 6-8 weeks old, weighing 32-42 g, Vital River, Beijing) as follows:

[0612] The pharmacokinetic profiles of the compounds following intravenous and oral administration in rodents were tested using standard protocols. Candidate compounds were formulated as clear solutions and administered to mice as single intravenous (IV) and oral (PO) injections. The vehicle consisted of 5% DMSO + 10% Solutol + 85% ddH2O. Oral administration groups fasted overnight (10-14 hours) prior to dosing. Whole blood samples were collected within 24 hours of dosing (0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours for oral administration; 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours for intravenous administration). Blood samples were collected from the submandibular vein or other appropriate means. Approximately 30 μL of blood was collected per time point, anticoagulated with K2-EDTA, and kept on ice until centrifugation. Blood samples were centrifuged at 4°C at 6800g for 6 minutes to obtain plasma. Plasma was stored at -80°C prior to measuring blood drug concentrations. LC-MS / MS was used to quantitatively analyze the blood drug concentration and calculate the pharmacokinetic parameters, such as peak concentration (C max ), distribution volume (V ss ) clearance (Cl), half-life (T 1 / 2 ), area under the drug-time curve (AUC 0-last ), bioavailability (F), etc. The test results are shown in Table 2.

[0613] Table 2 Pharmacokinetic data of the compounds of the present invention

[0614] As shown in Table 2, the compounds of the present disclosure exhibited favorable exposure, clearance, and bioavailability within the administered concentrations, doses, and timeframes. Upon intravenous administration, compound E019 exhibited a clearance (CL) of 8.97 mL / min / kg, significantly superior to the CL of Comparative Example 1 (27.05 mL / min / kg). Upon oral administration, the plasma exposures of compounds E018 and E019 were 7660.90 hr*ng / mL and 13343.43 hr*ng / mL, respectively, representing increases of 3.5-fold and 6-fold, respectively, compared to those of Comparative Example 1. Furthermore, the oral bioavailability of compounds E018 and E019 was more than 2-fold higher than that of Comparative Example 1. This demonstrates that the compounds of the present disclosure exhibit favorable pharmacokinetic properties and significantly outperform Comparative Example 1.

[0615] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substituted product thereof, in, X 1 N or CR 15 ; X 2 and X 3 are independently N or CH; Where X 1 , X 2 and X 3 At least one is N; R 1 For hydrogen, C 1-9 Alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 1 ; R 2 is hydrogen, -C(O)-R 7 、-C(O)OR 7 、-C(O)N(R 7 2. C 1-9 Alkyl, C 3-15 Cycloalkyl; R 3 is a heterocyclic group or a heteroaryl group, wherein each heterocyclic group or heteroaryl group optionally has 1 to 4 Z 3 ; R 4 is aryl, heterocyclic, or heteroaryl, wherein the aryl, heterocyclic, or heteroaryl is optionally substituted with 1 to 4 Z 4 ; R 5 For hydrogen, halogen, -CN, -NO2, -OR 7 、-N(R 8 )(R 9 )、-S(O)-R 7 、-S(O)2R 7 、-S(O)2N(R 7 )2. -C(O)R 7 、-OC(O)-R 7 、-C(O)OR 7 、-OC(O)OR 7 、-OC(O)N(R 10 )(R 11 )、-C(O)N(R 7 )2、-N(R 7 )C(O)(R 7 ), C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-9 Alkylthio, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-9 Alkylthio, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl and heteroaryl may be optionally substituted with 1 to 4 Z 5 ; R 6 is hydrogen, -C(O)-R 7 、-C(O)OR 7 、-C(O)N(R 7 2. C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 3-10 The heterocyclic, aryl or heteroaryl groups may be optionally substituted with 1 to 4 Z 6 ; Each R 7 are independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl, or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl and heteroaryl may be optionally substituted with 1 to 4 Z 7 ; R 8 and R 9 is independently hydrogen at each occurrence, -S(O)2-R 10 、-C(O)-R 10 、-C(O)OR 10 、-C(O)N(R 10 )(R 11) , C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 8 ; R 10 and R 11 is independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein each C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl may be optionally substituted with 1 to 4 Z 9 ; Every Z 1 , Z 2 , Z 3 , Z 4 , Z 5 , Z 6 , Z 7 , Z 8 and Z 9 are independently hydrogen, oxo, halogen, -NO2, -N3, -CN, thio, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-8 Haloalkyl, C 3-15 Cycloalkyl, heterocyclyl, aryl or heteroaryl, -OR 12 、-C(O)R 12 、-C(O)OR 12 、-C(O)N(R 13 )(R 14 )、-N(R 13 )(R 14 )、-N(R 13 )2(R 14 ) + 、-N(R 12 )C(O)-R 12 、-N(R 12 )C(O)OR 12 、-N(R 12 )C(O)N(R 13 )(R 14 )、-N(R 12 )S(O)2(R 12 ),-NR 12 S(O)2N(R 13 )(R 14 ),-NR 12 S(O)2O(R 12 )、-OC(O)R 12 、-OC(O)-N(R 13 )(R 14 )、-P(O)(OR 12 )2.-OP(O)(OR 12 )2、-CH2P(O)(OR 12 )2、-OCH2P(O)(OR 12 )2、-C(O)OCH2P(O)(OR 12 )2、-P(O)(R 12 )(OR 12 )、-OP(O)(R 12 )(OR 12 )、-CH2P(O)(R 12 )(OR 12 )、-OCH2P(O)(R 12 )(OR 12 )、-C(O)O-CH2P(O)(R 12 )(OR 12 )、-P(O)(N(R 12 )2)2、-OP(O)(N(R 12 )2)2、-CH2P(O)(N(R 12 )2)2、-OCH2P(O)-(N(R 12 )2)2、-C(O)OCH2P(O)(N(R 12 )2)2、-P(O)(N(R 12 )2)(OR 12 )、-OP(O)(N(R 12 )2)-(OR 12 )、-CH2P(O)(N(R 12 )2)(OR 12 )、-OCH2P(O)(N(R 12 )2)(OR 12 )、-C(O)OCH2P(O)(N(R 12 )2)(OR 12 )、-P(O)(R 12 )(N(R 12 )2)、-OP(O)(R 12 )(N(R 12 )2)、-CH2P(O)(R 12 )(N(R 12 )2)、-OCH2P(O)(R 12 )(N(R 12 )2)、-C(O)OCH2P(O)(R 12 )(N(R 12 )2)-SR 12 、-S(O)R 12 、-S(O)(NH)R 12 、-S(O)2R 12 Or -S(O)2N(R 13 )(R 14 ); wherein any alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, aryl, heteroaryl or heterocyclic group is optionally substituted by 1 to 4 substituents selected from the group consisting of oxo, halogen, hydroxyl, thio, -NO2, -CN, -N3, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, C 1-9 Alkoxy, C 1-9 Alkyl hydroxyl, -COOH, -COOC 1-9 Alkyl, amino, C 1-9 Alkylamino, (C 1-9 Alkyl)2amino, C 1-8 Haloalkyl, C 1-9 Alkylcarbonyl, -CONH2, -CON(C 1-9 alkyl) 1-2 C 1-9 Alkylsulfonyl, aryl, heteroaryl, heterocyclic; Each R 12 are independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, aryl, heterocyclic group, or heteroaryl group, wherein any alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl or heterocyclic group optionally has 1 to 4 Z 7 Group; R 13 and R 14 Each occurrence is independently hydrogen, C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-15 Cycloalkyl, heterocyclic, aryl or heteroaryl; any alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl or heterocyclic group optionally has 1 to 4 Z 8 group; or R 13 and R 14 Together with the nitrogen to which they are attached, they form a heterocyclic group, wherein the heterocyclic group is optionally substituted with 1 to 4 Z 8 groups; and Each R 15 are independently H, halogen, -CN, -NO2, -OR 7 、-N(R 8 )(R 9 )、-S(O)-R 7 、-S(O)2R 7 、-S(O)2N(R 7 )2. -C(O)R 7 、-OC(O)-R 7 、-C(O)OR 7 、-OC(O)OR 7 、-OC(O)N(R 10 )(R 11 )、-C(O)N(R 7 )2、-N(R 7 )C(O)(R 7 ), C 1-9 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-9 Alkylthio, C 1-6 Haloalkyl, C 3-15 Cycloalkyl, aryl, heterocyclyl or heteroaryl.

2. The compound according to claim 1, characterized in that R 1 Select from the following group:

3. The compound according to claim 1, characterized in that The compound has a structure shown in formula III-1, III-2 or III-3: Among them, each R 1 , R 3 , R 4 , R 5 and R 6 Independently as defined in claim 1.

4. The compound according to claim 1, characterized in that The compound has a structure shown in Formula IV-1, IV-2 or IV-3: Among them, each Z 3 , R 1 , R 4 , R 5 and R 6 Independently as defined in claim 1.

5. The compound according to claim 1, characterized in that The compound has a structure shown in Formula IV-1a, IV-2a or IV-3a: Where Z 3 , R 1 , R 4 , R 5 and R 6 Independently as defined in claim 1.

6. The compound according to claim 1, characterized in that R 3 Select from the following group:

7. The compound according to claim 1, characterized in that Z 3 Selected from 8. The compound according to claim 1, characterized in that R 4 Selected from 9. The compound according to claim 4, characterized in that The compound has a structure of formula IV-1, wherein Z 3 Selected from R 5 is halogen or cyano; R 1 Select from the following group: R 4 Select from the following group: and R 6 is selected from hydrogen, methyl, ethyl or cyclopropyl.

10. The compound according to claim 1, characterized in that The compound is selected from:

11. A pharmaceutical composition comprising the compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substitution thereof, and a pharmaceutically acceptable carrier.

12. Use of the compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, stereoisomer, stereoisomer mixture or isotope substitution thereof, or the pharmaceutical composition according to claim 2 in the preparation of a medicament for preventing or treating a disease or condition mediated by Osaka thyroid carcinoma (Cot) kinase.

13. The use according to claim 12, characterized in that The disease or condition is selected from the group consisting of cancer, diabetes, and inflammatory diseases.

14. The use according to claim 12, characterized in that The disease or condition is selected from the group consisting of type 1 and type 2 diabetes, non-alcoholic fatty liver disease (NAFLD) and inflammatory diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), inflammatory bowel disease (IBD), sepsis, psoriasis, dysregulated TNF expression and transplant rejection; acute gout and ankylosing spondylitis, allergic diseases, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis and multiple sclerosis, atherosclerosis, bacterial infection, bone cancer pain and pain caused by endometriosis; BR AF-resistant melanoma, colon cancer, brain stem glioma or pituitary adenoma, burns, bursitis, cancer of the anal region, cancer of the endocrine system, cancer of the kidney or ureter (e.g., renal cell carcinoma, renal pelvis), cancer of the penis, cancer of the small intestine, non-small cell lung cancer, cancer of the thyroid gland, cancer of the urethra, blood cancers such as acute myeloid leukemia, tongue cancer, cancer of the cervix, endometrium, fallopian tube, renal pelvis, vagina, or vulva, chronic myeloid leukemia, chronic or acute leukemia, chronic pain, classic Bartter syndrome syndrome), common cold, conjunctivitis, coronary heart disease, skin or intraocular melanoma, dermatitis, dysmenorrhea, eczema, endometriosis, familial adenomatous polyposis, fibromyalgia, fungal infection, gout, gynecological tumors, uterine sarcoma, fallopian tube cancer, headache, hemophilic arthropathy, Parkinson's disease, AIDS, herpes zoster, Hodgkin's disease, Huntington's disease, high prostaglandin E syndrome, influenza, iritis, juvenile arthritis, juvenile onset rheumatoid arthritis, juvenile rheumatoid arthritis, low back and neck pain, lymphocytic lymphoma, myofascial disorder, Myositis, neuralgia, neurodegenerative disorders such as Alzheimer's disease, neuritis, neuropathic pain, vulvar cancer, Parkinson's disease, pediatric malignancies, pulmonary fibrosis, colorectal cancer, rhinitis, sarcoidosis, soft tissue sarcoma, scleritis, skin cancer, solid tumors in children, spinal cord tumors, sprains and strains, gastric cancer, stroke, subacute and chronic musculoskeletal pain syndromes such as bursitis, surgical or dental procedures, symptoms associated with influenza or other viral infections, synovitis, toothache, ulcers, uterine cancer, uterine sarcoma, uveitis, vasculitis, viral infections, viral infections (e.g., influenza), and wound healing.

15. An intermediate compound represented by formula M1 in, R 16 Cl, Br, I, OTf and NH2; And X 1 , X 2 , X 3 , R 1 , R 2 and R 5 As defined in claim 1.

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