PARP1 and ULK1 dual-target inhibitor, and preparation method therefor and use thereof

Abstract

The present invention belongs to the field of pharmaceuticals, and particularly relates to a PARP1 and ULK1 dual-target inhibitor, and a preparation method therefor and the use thereof. A compound as represented by formula I or a pharmaceutically acceptable salt, a solvate, an active metabolite, a polymorph, an ester, an optical isomer or a prodrug thereof, and a composition comprising the compound as represented by formula I involved in the present invention all show a very strong inhibitory effect on PARP1 and ULK1. The compounds prepared in the present invention have a good inhibitory effect on many tumors that develop primary resistance to PARP1 inhibitors due to the autophagy induced by PARP1 inhibitors, such as triple-negative breast cancer, liver cancer and ovarian cancer.

Description

A PARP1 and ULK1 dual-target inhibitor and its preparation method and application Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a PARP1 and ULK1 dual-target inhibitor, and a preparation method and application thereof. Background Art

[0002] Poly(ADP-ribose) polymerase (PARP) is a multifunctional post-translational protein modification enzyme present in most eukaryotic cells. PARP family has 18 isoforms. Among them, PARP-1 accounts for the largest proportion and plays a major role in DNA damage repair [Peraltaleal A, et al., Free Radical Biology and Medicine 2009, 47(1):13-26].

[0003] Among DNA damage, the most serious damage is single-strand break (SSB) and double-strand break (DSB), among which single-strand break is more common. If these breaks cannot be repaired in time and accurately, the genome will become unstable, leading to cancer or directly leading to cell death. In order to maintain normal physiological functions, cells have a variety of DNA damage detection and repair mechanisms. The repair of single-strand break damage mainly depends on PARP, among which PARP1 plays more than 90% of the function [Langelier M, et al., Nucleic Acids Research 2014, 42(12):7762-7775]. As for double-strand break, although the occurrence is rare, the consequences are serious. If it cannot be repaired in time, the cell DNA will become unstable and eventually lead to death. Homologous recombination (HR) is a high-fidelity, error-free repair method for DNA double-strand break repair. The proteins involved in this repair include BRCA, ATM, RAD51, etc. The most well-known is BRCA protein. People carrying familial hereditary BRCA1 / 2 mutations have a significantly increased risk of malignant tumors, especially breast cancer and ovarian cancer. BRCA1 / 2 are key proteins involved in the homologous recombination repair of double-strand breaks. BRCA1 / 2 mutations impair DNA double-strand break repair, leading to genomic instability and, subsequently, cancer. Due to the cell's survival mechanism, impaired double-strand break repair makes cells more dependent on PARP-driven single-strand break repair. Compared to tumor cells with intact BRCA function, BRCA-deficient tumor cells are 100 times more sensitive to PARP inhibitors. The successful use of PARP inhibitors in BRCA-deficient cancer patients is a classic example of "synthetic lethality" in the DNA damage repair response (DDR) [Bryant H E., Nature 2005, 434(7035):913-917; Farmer H, et al., Nature 2005, 434(7035):917-921]. Currently, the PARP inhibitors that have been successfully marketed include Olaparib, Rucaparib, Niraparib, and Talazoparib.

[0004] Autophagy is a conserved intracellular degradation process that determines which organelles, proteins, and invading microorganisms are degraded by lysosomes. This conserved process is required for cellular responses to nutrient deprivation and other stresses, in addition to being required for proper cell and tissue homeostasis during embryonic development and for defense against pathogens. Defects in the autophagy pathway have been implicated in several human pathologies, including infectious diseases, neurodegenerative disorders, and cancer.

[0005] Autophagy is a multi-step pathway initiated by the ULK (unc-51-like autophagy-activating kinase) pre-initiation complex, which is composed of unc-51-like autophagy-activating kinase 1 (ULK1) or its homolog ULK2, autophagy-related protein 13 (ATG13), ATG101, and focal adhesion kinase (FAK) family interacting protein (FIP200) [Hosokawa N., et al., Autophagy 2009, 5:973-979; Ganley IG, et al., J Biol Chem 2009, 5:973-979]. Autophagy is activated by AMPK under stress conditions and inhibited by mTOR under non-stress conditions [Laplante M, et al., Cell 2012, 149:274-293]. AMPK activates autophagy through two mechanisms: first, it inhibits mTOR activity by phosphorylating the mTOR complex components raptor and TSC2; second, it activates ULK1 / 2 by directly phosphorylating multiple sites in the N-terminus of ULK1 / 2 to the mTOR phosphorylation site. Conversely, mTOR inhibits autophagy by directly phosphorylating and inhibiting ULK1 / 2 function.

[0006] As the only serine / threonine kinase in the autophagy signaling pathway, ULK1 / 2 plays a key role in autophagy by phosphorylating three proteins in the ULK1 / 2 pre-initiation complex (ATG13, ATG101 and FIP200)9-11 and the downstream Beclin1 initiation protein complex (Beclin1, VPS34, ATG9 and ATG16L1) [Papinski D, et al., Mol. Cell 2014, 53:471-483; Papinski D, et al., J Mol Biol 2016, 428:1725-1741; Egan D, et al., Mol Cell 2015, 59:285-297; Alsaadi R, EMBO Rep. 2019, 20:No.e46885].

[0007] Knocking out ULK1 or using small molecule ULK1 inhibitors can effectively inhibit autophagy, induce apoptosis, and inhibit tumor cell growth [Si-Tu Xue, et al., Autophagy 2020, 16(10): 1823-1837; Huiyu R, et al., J. Med. Chem. 2020, 63: 14609-14625].

[0008] Parp1 inhibitor-induced autophagy is the main cause of primary resistance to Parp1 inhibitors. The combined use of Parp1 inhibitors and autophagy inhibitors has shown strong synergistic anti-cancer effects in different tumors [Pai Bellare G, et al., Cancer 2021, 124: 1260-1274; Fu XT, et al., Cancer Cell Int. 2019, 19: 71; Luo T, et al., Autophagy 2016, 12: 1355-1371; Lu S, et al., Cell Death Dis. 2018, 9: 646; Liu Y, et al., AMB Expr 2019, 9: 108; Janice M SO, et al., Cancer 2020, 126: 894-907].

[0009] Therefore, developing a dual-target inhibitor that can simultaneously inhibit PARP1 and ULK1 will provide a new, safe and effective anti-cancer drug. Summary of the Invention

[0010] The technical problem to be solved by the present invention is to address the deficiencies of the existing technology and provide a novel compound that is a dual-target inhibitor of PARP1 and ULK1 and has not been reported in the literature, as well as pharmaceutically acceptable salts, solvates, active metabolites, polymorphs, esters, optical isomers or prodrugs thereof, the use of the compound in pharmaceutical manufacturing, and a method for using the compound of the present invention to prevent or treat diseases related to abnormal activity of PARP1 and ULK1 in humans or mammals.

[0011] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows:

[0012] The present invention discloses a compound represented by formula I or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof;

[0013] in,

[0014] R 1 Selected from halogen, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxy substituted C 1-6 Alkyl, alkoxy substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, halogenated C 3-6 Cycloalkyl or C 3-6 heterocycloalkyl;

[0015] R 2 Selected from-NR5 R 6 、-NHNR 5 R 6 、-NR 5 OR 6 、-OR 5 、-ONR 5 R 6 or -SR 5 ;in,

[0016] R 5 、R 6 are each independently selected from H, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted C 3-9 Heteroaryl, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 3-7 Heterocycloalkyl, or R 5 、R 6 with -NR 5 R 6 N in the form a 3-6 membered heterocyclic ring; wherein the substitution is selected from halogen, cyano, hydroxyl, amino, mono (C 1-3 alkyl)amino, di(C 1-3 Alkyl)amino, C 3-7 Cyclic amino group, C 3-7 Heterocyclic amino, C 4-7 Cyclic amide, C 1-3 Alkyl, halogenated C 1-3 Alkyl, C 1-3 Alkoxy or halogenated C 1-3 Alkoxy substitution;

[0017] R 3 Any one of the following groups:

[0018] Among them, R 4 Selected from H, halogen, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-3 Alkoxy or halogenated C 1-3 alkoxy;

[0019] L ring is selected from substituted or unsubstituted 5-6 membered aromatic rings, or substituted or unsubstituted 5-6 membered heteroaromatic rings; wherein the substitution is selected from halogen, cyano, hydroxyl, amino, mono(C 1-3 alkyl)amino, di(C 1-3 Alkyl)amino, C 1-3 Alkyl, halogenated C 1-3 Alkyl, C 1-3Alkoxy or halogenated C 1-3 Alkoxy substitution;

[0020] W is selected from N or CH.

[0021] Specifically, preferably, R 1 Selected from halogen, C 1-3 Alkyl, halogenated C 1-3 Alkyl, C 3-6 Cycloalkyl or C 3-6 Heterocycloalkyl.

[0022] Specifically, preferably, R 2 Selected from-NR 5 R 6 OR 5 ;in,

[0023] R 5 、R 6 are each independently selected from H, substituted or unsubstituted C6 aryl, substituted or unsubstituted C 3-5 Heteroaryl, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocycloalkyl, or R 5 、R 6 with -NR 5 R 6 N in the form a 3-6 membered heterocyclic ring; wherein the substitution is selected from halogen, cyano, hydroxyl, amino, mono (C 1-3 alkyl)amino, di(C 1-3 Alkyl)amino, C 4-7 Cyclic amide, C 1-3 Alkyl, halogenated C 1-3 Alkyl, C 1-3 Alkoxy or halogenated C 1-3 Alkoxy substitution.

[0024] Specifically, preferably, R 3 Any one selected from the following groups:

[0025] Among them, R 4 is selected from H or halogen.

[0026] Specifically, preferably, the L ring is selected from a substituted or unsubstituted 5-6 membered aromatic ring, or a substituted or unsubstituted 5-6 membered heteroaromatic ring; wherein the substitution is selected from halogen substitution.

[0027] Specifically, preferably, W is selected from N.

[0028] Specifically, preferably:

[0029] R 1 Selected from halogenated C 1-3 alkyl;

[0030] R 2 Selected from-NR 5 R 6 ;in,

[0031] R 5 、R 6 are each independently selected from H, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocycloalkyl; wherein the substitution is selected from amino or pyrrolidone;

[0032] R 3 Any one selected from the following groups:

[0033] Among them, R 4 Selected from H or fluorine;

[0034] The L ring is selected from a substituted or unsubstituted 6-membered aromatic ring, or an unsubstituted 6-membered heteroaromatic ring; wherein the substitution is selected from fluorine substitution;

[0035] W is selected from N;

[0036] Specifically, it is further preferred that:

[0037] R 1 Selected from trifluoromethyl;

[0038] R 2 Any one selected from the following groups:

[0039] R 3 Any one selected from the following groups:

[0040] The L ring is selected from any one of the following groups:

[0041] W is selected from N.

[0042] Specifically, more preferably, the compound is selected from any of the following structures:

[0043] Furthermore, the present invention provides a method for preparing the above-mentioned compound, characterized in that it comprises the following steps:

[0044] Step 1: Compound A-1 undergoes nucleophilic substitution with compound B to obtain compound A-2;

[0045] Step 2: Compound A-2 is coupled with compound C to generate compound I;

[0046] Wherein, the compound B is selected from amine, hydroxylamine, hydrazine, alcohol or thiol; the compound C is

[0047] in,

[0048] R 1 Selected from halogen, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxy substituted C 1-6 Alkyl, alkoxy substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, halogenated C 3-6 Cycloalkyl or C 3-6 heterocycloalkyl;

[0049] R 2 Selected from-NR 5 R 6 、-NHNR 5 R 6 、-NR 5 OR 6 、-OR 5 、-ONR 5 R 6 or -SR 5 ;in,

[0050] R 5 、R 6 are each independently selected from H, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted C 3-9 Heteroaryl, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 3-7 Heterocycloalkyl, or R 5 、R 6 with -NR 5 R 6 N in the form a 3-6 membered heterocyclic ring; wherein the substitution is selected from halogen, cyano, hydroxyl, amino, mono (C 1-3 alkyl)amino, di(C 1-3 Alkyl)amino, C 3-7 Cyclic amino group, C 3-7 Heterocyclic amino, C 4-7 Cyclic amide, C 1-3 Alkyl, halogenated C 1-3 Alkyl, C 1-3Alkoxy or halogenated C 1-3 Alkoxy substitution;

[0051] R 3 Any one of the following groups:

[0052] Among them, R 4 Selected from H, halogen, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-3 Alkoxy or halogenated C 1-3 alkoxy;

[0053] L ring is selected from substituted or unsubstituted 5-6 membered aromatic rings, or substituted or unsubstituted 5-6 membered heteroaromatic rings; wherein the substitution is selected from halogen, cyano, hydroxyl, amino, mono(C 1-3 alkyl)amino, di(C 1-3 Alkyl)amino, C 1-3 Alkyl, halogenated C 1-3 Alkyl, C 1-3 Alkoxy or halogenated C 1-3 Alkoxy substitution;

[0054] W is selected from N or CH.

[0055] Specifically, in step 1, compound A-1 and compound B undergo nucleophilic substitution to obtain compound A-2; wherein, the molar ratio of compound A-1 to compound B is 1:(0.80-4.00), preferably 1:(0.80-3.00), and more preferably 1:(0.90-2.00); the reaction temperature of the nucleophilic substitution is room temperature; preferably, the solvent used in the reaction process of the nucleophilic substitution is methanol or dichloromethane; a first base is added to the nucleophilic substitution reaction for catalysis; the first base is a trialkyl tertiary amine; preferably, the trialkyl tertiary amine is N,N-diisopropylethylamine; the molar ratio of compound A-1 to the first base is 1:(1.50-4.00), preferably 1:(1.50-3.50), and more preferably 1:(1.90-2.46); wherein, when the compound B is cyclopropylamine, the first base does not need to be added to the nucleophilic substitution reaction system.

[0056] Specifically, in step 2, compound A-2 is coupled with compound C to form compound I; wherein the molar ratio of compound A-2 to compound C is 1:(0.80-4.00), preferably 1:(0.80-2.00), and more preferably 1:(0.80-1.20); the coupling catalytic system for coupling compound A-2 and compound C to form compound I is one of the following catalytic systems:

[0057] Catalytic system 1: a catalytic system containing a palladium catalyst, an organophosphorus ligand, and a second base; wherein, preferably, the palladium catalyst is tris(dibenzylideneacetone)dipalladium; preferably, the organophosphorus ligand is 4,5-bisdiphenylphosphine-9,9-dimethylxanthene; preferably, the second base is cesium carbonate; the molar ratio of the compound A-2 to the palladium catalyst, the organophosphorus ligand, and the second base is 1:(0.08~0.40):(0.08~0.80):(1.50~6.00), preferably 1:(0.08~0.35):(0.08~0.60):(1.80~4.00), and more preferably 1:(0.08~0.25):(0.08~0.45):(2.00~3.20).

[0058] Among them, when the catalytic system used in the coupling process is catalytic system 1, the solvent used in the coupling process includes but is not limited to 1,4-dioxane, isopropanol and N,N-dimethylformamide, preferably 1,4-dioxane, and the coupling reaction is carried out under the protection of an inert gas. The reaction temperature of the coupling reaction is 100-140°C, preferably 125-135°C, and more preferably 130°C; wherein the inert gas is nitrogen.

[0059] or,

[0060] Catalytic system 2: a catalytic system containing a first acid; wherein the first acid is methanesulfonic acid, p-toluenesulfonic acid, trifluoroacetic acid or hydrochloric acid; when the first acid is methanesulfonic acid, p-toluenesulfonic acid or trifluoroacetic acid, the molar ratio of the compound A-2 to the first acid is 1:(0.60-1.20), preferably 1:(0.70-1.00), and more preferably 1:(0.80-0.95); when the first acid is hydrochloric acid, the first acid is used in an excess amount, preferably, the molar ratio of the compound A-2 to the first acid is 1:18-24.

[0061] Among them, when the catalytic system used in the coupling process is catalytic system 2, the solvent used in the coupling process includes but is not limited to N,N-dimethylformamide, 1,4-dioxane and isopropanol, and the reaction temperature of the coupling reaction is 80-140°C, preferably 90-120°C.

[0062] Specifically, in step 2, compound A-2 is coupled with compound C to generate compound I; when compound C R in 3 Selected from When R 3 middle The N atom on the upper side has high activity, and the N atom is protected with a tert-butyl group. After the coupling reaction is completed, it is deprotected under the action of a second acid to obtain compound I; preferably, the second acid is trifluoroacetic acid; the deprotection reaction temperature is 80-100°C, preferably 85-100°C, and more preferably 90-100°C.

[0063] Specifically, step 1: compound A-1 and compound B undergo nucleophilic substitution to obtain compound A-2; step 2: compound A-2 and compound C undergo coupling to generate compound I; when compound A-2 R in 2 Selected from When the N atom on the heterocycloalkyl group or the N atom on the substituted amino group on the cycloalkyl group is highly reactive, the N atom is protected with a tert-butyloxycarbonyl (Boc) group. After the coupling reaction is completed, the compound I is obtained by deprotection under the action of a third acid. Preferably, the third acid is trifluoroacetic acid or hydrochloric acid. The deprotection reaction temperature is room temperature.

[0064] Each product obtained by the reaction of the above method can be obtained by conventional separation techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc. The starting materials required for the synthesis can be synthesized in-house or purchased from commercial institutions such as, but not limited to, Adrich or Sigma. These raw materials can be characterized using conventional means, such as physical constants and spectral data. The compounds described in the present invention can be obtained using synthetic methods to obtain single optical isomers or mixtures of optical isomers.

[0065] In the present invention, the superscript letter represents the group number, and the subscript represents the number of the atom, for example: R 1 、R 2 、R 3 Indicates the 1st to 3rd R groups, C 1-4 Alkyl refers to an alkyl group containing 1 to 4 carbon atoms. The carbon atoms in the substituent are not counted in the main chain.

[0066] Furthermore, the present invention provides a pharmaceutical composition comprising the compound represented by the above-mentioned formula I or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0067] Furthermore, the present invention provides a pharmaceutical preparation comprising a therapeutically effective amount of the compound of Formula I above or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

[0068] The pharmaceutical preparation is formulated for an administration route selected from oral administration, parenteral administration, buccal administration, nasal administration, topical administration or rectal administration.

[0069] Furthermore, the use of the above-mentioned compound, pharmaceutical composition or pharmaceutical preparation in the preparation of poly(ADP-ribose) polymerase 1 inhibitors is also within the scope of protection of the present invention.

[0070] Furthermore, the use of the above-mentioned compound, the above-mentioned pharmaceutical composition or the above-mentioned pharmaceutical preparation in the preparation of UNC-51-like autophagy-activated kinase 1 inhibitors is also within the protection scope of the present invention.

[0071] Furthermore, the use of the above-mentioned compound, the above-mentioned pharmaceutical composition or the above-mentioned pharmaceutical preparation in the preparation of a drug for preventing or treating diseases related to abnormal poly(ADP-ribose) polymerase 1 and / or UNC-51-like autophagy-activated kinase 1 enzyme activity is also within the scope of protection of the present invention.

[0072] Among them, the poly (ADP-ribose) polymerase 1 is abbreviated as Parp1; the unc-51-like autophagy-activating kinase 1 is abbreviated as ULK1.

[0073] Specifically, the disease associated with abnormal activity of poly(ADP-ribose) polymerase 1 and / or unc-51-like autophagy-activated kinase 1 is a tumor.

[0074] Specifically, preferably, the tumor includes a solid tumor or a malignant blood tumor; further preferably, the solid tumor is a breast cancer tumor or a liver cancer tumor.

[0075] Among them, the above-mentioned compound or the above-mentioned pharmaceutical composition or the above-mentioned pharmaceutical preparation is used to prevent or treat diseases or conditions related to abnormal activity of poly(ADP-ribose) polymerase 1 and / or UNC-51-like autophagy-activated kinase 1 enzymes, including administering the above-mentioned compound or the above-mentioned pharmaceutical composition or the above-mentioned pharmaceutical preparation to a human or mammal in need thereof, and the disease related to abnormal activity of poly(ADP-ribose) polymerase 1 and / or UNC-51-like autophagy-activated kinase 1 enzymes is cancer.

[0076] The present invention includes the step of contacting the compound, the pharmaceutical composition or the pharmaceutical preparation with PARP1 / ULK1, and the contacting step includes in vitro or in vivo testing. Beneficial effects:

[0077] The compounds prepared in this application show strong inhibitory effects on both PARP1 and ULK1. The compounds prepared in this application will have good inhibitory effects on many tumors that are primarily resistant to Parp1 inhibitors due to autophagy induced by Parp1 inhibitors, such as triple-negative breast cancer, liver cancer, ovarian cancer, etc. DETAILED DESCRIPTION

[0078] The present invention can be better understood according to the following examples. However, it is easy for those skilled in the art to understand that the contents described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0079] Example 1: Synthesis of Intermediate A-2-1

[0080] Synthesis route of 2-chloro-N-cyclopropyl-5-(trifluoromethyl)pyrimidin-4-amine (Compound A-2-1):

[0081] Compound B-1 (649 mg, 11.3 mmol) was added to a methanol (20 mL) solution containing compound A-1-1 (1.447 g, 6.7 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 4 hours; after the reaction, the reaction solution was poured into water (50 mL) and extracted twice with ethyl acetate (2 x 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:20 to obtain product A-2-1, 200 mg, with a yield of 12.6%.

[0082] The H NMR spectrum data of product A-2-1 are: 1 H NMR (400MHz, CDCl3): δ=8.27(s,1H); 7.27(s,1H); 2.93-2.99(m,1H); 0.90-1.02(m,2H); 0.60-0.68(m,2H).

[0083] Example 2: Synthesis of Intermediate A-2-2

[0084] Synthesis route of 2-chloro-N-cyclobutyl-5-(trifluoromethyl)pyrimidin-4-amine (Compound A-2-2):

[0085] To a DCM (30 mL) solution containing compound A-1-1 (3.0 g, 13.9 mmol) and DIEA (4.41 g, 34.2 mmol) was added compound B-2 (1.0 g, 14.0 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:25 to obtain product A-2-2, 1.7 g, with a yield of 48.6%.

[0086] The H NMR spectrum data of product A-2-2 are: 1 H NMR (400MHz, CDCl3): δ=8.24(s,1H); 5.49(s,1H); 4.63-4.69(m,1H); 2.44-2.52(m,2H); 1.90-1.97(m,2H); 1.78-1.85(m,2H).

[0087] Example 3: Synthesis of Intermediate A-2-3

[0088] Synthesis route of 2-chloro-N-(oxetane-3-yl)-5-(trifluoromethyl)pyrimidin-4-amine (Compound A-2-3):

[0089] To a DCM (30 mL) solution containing compound A-1-1 (3.0 g, 13.9 mmol) and DIEA (4.41 g, 34.2 mmol) was added compound B-3 (1.0 g, 13.7 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:25 to obtain product A-2-3, 0.88 g, with a yield of 25.0%.

[0090] The H NMR spectrum data of product A-2-3 are: 1 H NMR (400MHz, CDCl3): δ = 8.33 (s, 1H); 5.84 (s, 1H); 5.22-5.30 (m, 1H); 5.01-5.06 (m, 2H); 4.57-4.60 (m, 2H).

[0091] Example 4: Synthesis of Intermediate A-2-5

[0092] Synthesis route of tert-butyl (1R, 4R)-4-(2-chloro-5-(trifluoromethyl)pyrimidin-4-ylamino)cyclohexylcarbamate (Compound A-2-5):

[0093] Compound B-5 (1.0 g, 4.6 mmol) was added to a DCM (10 mL) solution containing compound A-1-1 (1.0 g, 4.6 mmol) and DIEA (1.2 g, 9.2 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:5 to obtain product A-2-5, 1.1 g, with a yield of 60.7%.

[0094] The H NMR spectrum data of product A-2-5 are: 1 H NMR (400MHz, CDCl3): δ = 8.17 (s, 1H); 5.07-5.09 (m, 1H); 4.30-4.40 (m, 1H); 3.90-4 .10(m,1H); 3.30-3.50(m,1H); 1.95-2.10(m,4H); 1.15-1.30(m,4H); 1.52(s,9H).

[0095] Example 5: Synthesis of Intermediate A-2-6

[0096] Synthesis route of 2-chloro-N-(tetrahydrofuran-3-yl)-5-(trifluoromethyl)pyrimidin-4-amine (Compound A-2-6):

[0097] To a DCM (30 mL) solution containing compound A-1-1 (2.5 g, 11.6 mmol) and DIEA (2.96 g, 23.0 mmol) was added compound B-6 (1.0 g, 11.5 mmol, Mw = 87.122) at 0 ° C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:5 to obtain product A-2-6, 0.39 g, with a yield of 12.6%.

[0098] The H NMR spectrum data of product A-2-6 are: 1H NMR (400MHz, CDCl3): δ = 8.29 (s, 1H); 5.51 (s, 1H); 4.80-4.90 (m, 1H); 3.90-4.10 (m ,2H); 3.80-3.90(m,1H); 3.70-3.80(m,1H); 2.30-2.50(m,1H); 1.80-1.90(m,1H).

[0099] Example 6: Synthesis of Intermediate A-2-7

[0100] Synthesis route of 2-chloro-N-(tetrahydro-2H-pyran-4-yl)-5-(trifluoromethyl)pyrimidin-4-amine (Compound A-2-7):

[0101] Compound B-7 (1.0 g, 10.0 mmol) was added to a DCM (30 mL) solution containing compound A-1-1 (2.2 g, 10.2 mmol) and DIEA (2.6 g, 20.0 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:5 to obtain product A-2-7, 1.3 g, with a yield of 45.4%.

[0102] The H NMR spectrum data of product A-2-7 are: 1 H NMR (400MHz, CDCl3): δ=8.20(s,1H); 5.15(s,1H); 4.20-4.40(m,1H); 3.90-4.00(m,2H); 3.50-3.70(m,2H); 1.90-2.00(m,2H); 1.40-1.60(m,2H).

[0103] Example 7: Synthesis of Intermediate A-2-8

[0104] Synthesis route of tert-butyl 4-(2-chloro-5-(trifluoromethyl)pyrimidin-4-ylamino)piperidine-1-carboxylate (Compound A-2-8):

[0105] To a DCM (10 mL) solution containing compound A-1-1 (1.09 g, 5.0 mmol) and DIEA (1.3 g, 10.0 mmol) was added compound B-8 (1.0 g, 5.0 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:10 to obtain product A-2-8, 0.47 g, with a yield of 24.7%.

[0106] The H NMR spectrum data of product A-2-8 are: 1 H NMR (400MHz, CDCl3): δ = 8.27 (s, 1H); 5.20 (m, 1H); 4.20-4.40 (m, 1H); 3.95-4.1 5(m,2H); 2.90-3.00(m,2H); 1.95-2.10(m,2H); 1.62(s,9H); 1.30-1.50(m,2H).

[0107] Example 8: Synthesis of Intermediate A-2-9

[0108] Synthesis route of tert-butyl 3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-ylamino)pyrrolidine-1-carboxylate (Compound A-2-9):

[0109] To a DCM (12 mL) solution containing compound A-1-1 (1.16 g, 5.37 mmol) and DIEA (1.38 g, 10.74 mmol) was added compound B-9 (1.0 g, 5.37 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:7 to obtain product A-2-9, 0.93 g, with a yield of 47.3%.

[0110] The H NMR spectrum data of product A-2-9 are: 1H NMR (400MHz, CDCl3): δ = 8.23 ​​(s, 1H); 5.30 (m, 1H); 4.65-4.75 (m, 1H); 3.65-3.75 (m, 1H); 3.35-3.50(m,2H); 3.10-3.30(m,1H); 2.20-2.35(m,1H); 1.80-1.90(m,1H); 1.41(s,9H).

[0111] Example 9: Synthesis of Intermediate A-2-10

[0112] Synthesis route of tert-butyl (1S,3S)-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-ylamino)cyclopentylcarbamate (Compound A-2-10):

[0113] To a DCM (5 mL) solution containing compound A-1-1 (0.27 g, 1.25 mmol) and DIEA (0.32 g, 2.5 mmol) was added compound B-10 (0.25 g, 1.25 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (10 mL) and extracted three times with ethyl acetate (3 x 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:3 to obtain product A-2-10, 0.24 g, with a yield of 50.5%.

[0114] The H NMR spectrum data of product A-2-10 are: 1 H NMR (400MHz, CDCl3): δ = 8.26 (s, 1H); 5.33 (m, 1H); 4.50-4.70 (m, 2H); 4.00-4.15 (m, 1H); 2.30-2.4 0(m,1H); 2.15-2.30(m,1H); 2.00-2.10(m,1H); 1.85-2.00(m,1H); 1.40-1.6(m,2H); 1.49(s,9H).

[0115] Example 10: Synthesis of Intermediate A-2-11

[0116] Synthesis route of 1-(3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-ylamino)propyl)pyrrolidin-2-one (Compound A-2-11):

[0117] To a DCM (15 mL) solution containing compound A-1-1 (1.53 g, 7.08 mmol) and DIEA (1.8 g, 14.0 mmol) was added compound B-11 (1.0 g, 7.04 mmol) at 0°C to obtain a mixed solution; the mixed solution was warmed to room temperature and stirred for 12 hours; after the reaction was completed, the reaction solution was poured into water (30 mL) and extracted three times with ethyl acetate (3 x 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography with ethyl acetate / petroleum ether = 1:1 to obtain product A-2-11, 1.23 g, with a yield of 53.9%.

[0118] The H NMR spectrum data of product A-2-11 are: 1 H NMR (400MHz, CDCl3): δ = 8.15 (s, 1H); 7.28 (m, 1H); 3.44-3.49 (m, 2H); 3.30-3.40 (m ,2H); 3.25-2.30(m,2H); 2.38-2.42(m,2H); 1.95-2.10(m,2H); 1.65-1.75(m,2H).

[0119] Example 11: Synthesis of Intermediate C-1

[0120] Synthesis route of compound C-1:

[0121] (1) Synthesis of 2-(4-nitrophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound M-3)

[0122] Compound M-1 (0.86 g, 5.70 mmol), M-2 (1 g, 6.17 mmol), HATU (1.4 g, 3.68 mmol), and DIEA (0.88 g, 6.79 mmol) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction, 60 mL of water was added to the reaction mixture, filtered, and the filter cake dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 hours. After the reaction, the mixture was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (2 x 40 mL). The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to obtain product M-3, 0.92 g, in a yield of 57.2%. LCMS of product M-3: m / z = 282.9 [M+1] + .

[0123] (2) Synthesis of 2-(4-aminophenyl)-1H-benzo[d]imidazole-4-carboxamide (C-1)

[0124] M-3 (0.4 g, 1.42 mmol) prepared in step (1), Fe (0.395 g, 7.1 mmol), NH4Cl (0.385 g, 7.1 mmol), EtOH (6 mL), and H2O (1.2 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, and 8 mL of water was added. The solution was extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (eluted with EA) to obtain product C-1, 0.3 g, with a yield of 83.8%. LCMS of product C-1: m / z = 252.9 [M+1] + .

[0125] Example 12: Synthesis of Intermediate C-2

[0126] Synthesis route of compound C-2:

[0127] (1) Synthesis of 2-(3-nitrophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound M-5)

[0128] Compounds M-1 (0.86 g, 5.7 mmol), M-4 (1 g, 6.17 mmol), HATU (1.4 g, 3.68 mmol), and DIEA (0.88 g, 6.79 mmol) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction, 60 mL of water was added to the reaction mixture, filtered, and the filter cake dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 hours. After the reaction, the mixture was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (2 x 40 mL). The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to obtain product M-5, 0.95 g, in a yield of 59.1%. LCMS of product M-5: m / z = 282.9 [M+1] + .

[0129] (2) Synthesis of 2-(3-aminophenyl)-1H-benzo[d]imidazole-4-carboxamide (C-2)

[0130] M-5 (0.4 g, 1.42 mmol), Fe (0.395 g, 7.1 mmol), NH4Cl (0.385 g, 7.1 mmol), EtOH (6 mL), and H2O (1.2 mL) prepared in step (1) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, 8 mL of water was added, and extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (EA elution) to obtain product C-2, 0.28 g, with a yield of 78.2%.

[0131] LCMS of product C-2: m / z=253.0[M+1] + .

[0132] Example 13: Synthesis of Intermediate C-3

[0133] Synthesis route of compound C-3:

[0134] (1) Synthesis of 2-(2-fluoro-4-nitrophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound M-7)

[0135] Compounds M-1 (0.86 g, 5.7 mmol), M-6 (1 g, 5.41 mmol), HATU (1.4 g, 3.68 mmol), and DIEA (0.88 g, 6.79 mmol) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction, 60 mL of water was added to the reaction mixture, filtered, and the filter cake dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 hours. After the reaction, the mixture was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (2 x 40 mL). The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to obtain product M-7, 0.89 g, in a yield of 52.0%. LCMS of product M-7: m / z = 300.9 [M+1] + .

[0136] (2) Synthesis of 2-(4-amino-2-fluorophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound C-3)

[0137] M-7 (0.43 g, 1.43 mmol) prepared in step (1), Fe (0.395 g, 7.1 mmol), NH4Cl (0.385 g, 7.1 mmol), EtOH (6 mL), and H2O (1.2 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, and 8 mL of water was added. The solution was extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (eluted with EA) to obtain product C-3, 0.28 g, with a yield of 72.5%. LCMS of product C-3: m / z = 270.9 [M+1] + .

[0138] Example 14: Synthesis of Intermediate C-4

[0139] Synthesis route of compound C-4:

[0140] (1) Synthesis of 2-(2-fluoro-5-nitrophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound M-9)

[0141] Compound M-1 (0.77 g, 5.10 mmol), M-8 (1 g, 5.41 mmol), HATU (1.23 g, 3.24 mmol), and DIEA (0.76 g, 5.94 mmol) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction, 60 mL of water was added to the reaction mixture, filtered, and the filter cake dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 hours. After the reaction, the mixture was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (2 x 40 mL). The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to obtain product M-9, 0.87 g, in a yield of 56.9%. LCMS of product M-9: m / z = 300.9 [M+1] + .

[0142] (2) Synthesis of 2-(5-amino-2-fluorophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound C-4)

[0143] M-9 (0.43 g, 1.43 mmol) prepared in step (1), Fe (0.395 g, 7.1 mmol), NH4Cl (0.385 g, 7.1 mmol), EtOH (6 mL), and H2O (1.2 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, and 8 mL of water was added. The solution was extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (eluted with EA) to obtain product C-4, 0.28 g, with a yield of 72.5%. LCMS of product C-4: m / z = 270.9 [M+1] + .

[0144] Example 15: Synthesis of Intermediate C-5

[0145] Synthesis route of compound C-5:

[0146] (1) Synthesis of 2-(5-nitropyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (Compound M-11)

[0147] Compounds M-1 (0.86 g, 5.7 mmol), M-10 (1 g, 5.95 mmol), HATU (1.4 g, 3.57 mmol), and DIEA (0.85 g, 6.55 mmol) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction, 60 mL of water was added to the reaction mixture, filtered, and the filter cake dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 hours. After the reaction, the mixture was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (2 x 40 mL). The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to obtain product M-11, 0.83 g, in a yield of 51.5%. LCMS of product M-11: m / z = 305.8 [M+23]. + .

[0148] (2) Synthesis of 2-(5-aminopyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (Compound C-5)

[0149] M-11 (0.4 g, 1.41 mmol) prepared in step (1), Fe (0.395 g, 7.1 mmol), NH4Cl (0.385 g, 7.1 mmol), EtOH (6 mL), and H2O (1.2 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, and 8 mL of water was added. The solution was extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (eluted with EA) to obtain product C-5, 0.11 g, with a yield of 30.8%. LCMS of product C-5: m / z = 253.9 [M+1] + .

[0150] Example 16: Synthesis of Intermediate C-6

[0151] Synthesis route of compound C-6:

[0152] (1) Synthesis of 2-(5-nitropyridin-3-yl)-1H-benzo[d]imidazole-4-carboxamide (Compound M-13)

[0153] Compound M-1 (0.86 g, 5.7 mmol), M-12 (1 g, 5.95 mmol), HATU (1.4 g, 3.57 mmol), and DIEA (0.85 g, 6.55 mmol) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction, 60 mL of water was added to the reaction mixture, filtered, and the filter cake dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 hours. After the reaction, the mixture was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (2 x 40 mL). The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to obtain product M-13, 0.75 g, in a yield of 46.5%. LCMS of product M-13: m / z = 305.8 [M+23]. + .

[0154] (2) Synthesis of 2-(5-aminopyridin-3-yl)-1H-benzo[d]imidazole-4-carboxamide (Compound C-6)

[0155] M-13 (0.4 g, 1.41 mmol) prepared in step (1), Fe (0.395 g, 7.1 mmol), NH4Cl (0.385 g, 7.1 mmol), EtOH (6 mL), and H2O (1.2 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, and 8 mL of water was added. The solution was extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (eluted with EA) to obtain product C-6, 0.12 g, with a yield of 33.6%. LCMS of product C-6: m / z = 253.9 [M+1] + .

[0156] Example 17: Synthesis of Intermediate C-7

[0157] Synthesis route of compound C-7:

[0158] (1) Synthesis of N-tert-butyl-2-(4-nitrophenyl)-2H-indazole-7-carboxamide (Compound M-16)

[0159] M-14 (5.0 g, 23.04 mmol), M-15 (3.25 g, 23.04 mmol), and K2CO3 (6.36 g, 46.08 mmol) were dissolved in DMF (70 mL) and refluxed at 200°C for 2 h. After the reaction, the reaction solution was cooled to room temperature, 60 mL of water was added, filtered, and the filter cake was dried. The filter cake was added to 12 mL of AcOH and refluxed at 120°C for 4 h. After the reaction, the solution was cooled to room temperature, 60 mL of water was added, and the solution was extracted with EA (2*40 mL). The organic phase was dried, concentrated, and dried to give the product M-16 (4.0 g).

[0160] (2) Synthesis of 2-(4-aminophenyl)-N-tert-butyl-2H-indazole-7-carboxamide (Compound C-7)

[0161] M-16 (0.51 g, 1.5 mmol) prepared in step (1), Fe (0.42 g, 7.5 mmol), NH4Cl (0.41 g, 7.5 mmol), EtOH (10 mL), and H2O (2 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, 8 mL of water was added, and extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (eluted with EA) to obtain product C-7, 0.37 g. LCMS of product C-7: m / z=310.0 [M+1] + .

[0162] Example 18: Synthesis of Intermediate C-8

[0163] Synthesis route of compound C-8:

[0164] (1) Synthesis of N-tert-butyl-2-(3-nitrophenyl)-2H-indazole-7-carboxamide (Compound M-19)

[0165] M-14 (1.0 g, 4.6 mmol), M-18 (1.25 g, 7.5 mmol), Cu(OAc)2 (6.36 g, 31.9 mmol), and Py (12.43 mmol, 1 mL) were dissolved in DMF (70 mL) and refluxed at 80°C in air for 12 h. After the reaction, the reaction solution was cooled to room temperature, 60 mL of water was added, and the mixture was extracted with EA (3 x 100 mL). The organic phase was dried and concentrated. The product was purified by column chromatography and dried to give M-19 (0.6 g).

[0166] (2) Synthesis of 2-(3-aminophenyl)-N-tert-butyl-2H-indazole-7-carboxamide (C-8)

[0167] M-19 (0.338 g, 1.0 mmol) prepared in step (1), Fe (0.28 g, 5.0 mmol), NH4Cl (0.27 g, 5.0 mmol), EtOH (5 mL), and H2O (1 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, 8 mL of water was added, and extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (PE:EA=1:1) to obtain product C-8, 0.2 g. LCMS of product C-8: m / z=309.0 [M+1] + .

[0168] Example 19: Synthesis of Intermediate C-9

[0169] Synthesis route of compound C-9:

[0170] (1) Synthesis of 2-(4-nitrophenyl)-8-fluoro-4,5-dihydro-1H-aza[5,4,3-cd]indol-6(3H)-one (Compound M-23)

[0171] M-21 (0.4 g, 1.42 mmol), M-22 (0.237 g, 1.42 mmol), PdCl2(dppf) (0.1 g, 0.14 mmol), and Na2CO3 (300 mg, 2.84 mmol, 2 eq) were dissolved in dioxane (7 mL) and refluxed at 90°C for 2 h. After completion of the reaction, the reaction solution was cooled to room temperature, 20 mL of water was added, and the mixture was extracted with EA (3 x 30 mL). The organic phase was dried and concentrated. Purification by column chromatography afforded the product M-23 (0.21 g).

[0172] (2) Synthesis of 2-(4-aminophenyl)-8-fluoro-4,5-dihydro-1H-aza[5,4,3-cd]indol-6(3H)-one (C-9)

[0173] M-23 (0.21 g, 0.65 mmol) prepared in step (1), Fe (0.18 g, 3.35 mmol), NH4Cl (0.179 g, 3.35 mmol), EtOH (5 mL), and H2O (1 mL) were mixed and refluxed at 70°C for 4 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, and 8 mL of water was added. The mixture was extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (DCM:MeOH=8:1) to obtain product C-9, 0.08 g. LCMS of product C-9: m / z=296.0 [M+1] + .

[0174] Example 20: Synthesis of Intermediate C-10

[0175] Synthesis route of compound C-10:

[0176] (1) Synthesis of 2-(3-nitrophenyl)-8-fluoro-4,5-dihydro-1H-aza[5,4,3-cd]indol-6(3H)-one (Compound M-25)

[0177] M-21 (0.4 g, 1.41 mmol), M-24 (0.237 g, 1.42 mmol), PdCl2(dppf) (0.1 g, 0.14 mmol), and Na2CO3 (300 mg, 2.84 mmol, 2 eq) were dissolved in dioxane (7 mL) and refluxed at 90°C for 2 h. After completion of the reaction, the reaction solution was cooled to room temperature, 20 mL of water was added, and the mixture was extracted with EA (3 x 30 mL). The organic phase was dried and concentrated. Purification by column chromatography afforded the product M-25 (0.24 g).

[0178] (2) Synthesis of 2-(3-aminophenyl)-8-fluoro-4,5-dihydro-1H-aza[5,4,3-cd]indol-6(3H)-one (Compound C-10)

[0179] M-25 (0.13 g, 0.4 mmol) prepared in step (1), Fe (0.112 g, 2.0 mmol), NH4Cl (0.108 g, 2.0 mmol), EtOH (3 mL), and H2O (0.6 mL) were mixed and refluxed at 70°C for 6 hours. After the reaction, the reaction solution was cooled to room temperature, concentrated, 8 mL of water was added, and extracted with EA (3*20 mL). The organic phase was dried, mixed with silica gel, and purified by column chromatography (PE:EA=1:5) to obtain product C-10, 0.018 g. LCMS of product C-10: m / z=296.0 [M+1] + .

[0180] Example 21: Synthesis of Intermediate C-11

[0181] Synthesis route of compound C-11:

[0182] (1) Synthesis of 7-fluoro-9-nitro-1,2,3,4-tetrahydrobenzo[e][1,4]diazepin-5-one (Compound M-28)

[0183] A mixture containing M-26 (1.5 g, 5.40 mmol), M-27 (1.07 g, 8.01 mmol), and triethylamine (3 mL, 21.58 mmol) in DMA (6 mL) was stirred at 100°C for 1 h. After completion, the reaction mixture was cooled, diluted with EA (100 mL), and washed with water (2 x 50 mL) and saturated NaCl solution (2 x 20 mL). The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography using PE / EA (2:1) to afford the product M-28, 0.8 g, in a yield of 65.8%. LCMS of the product M-28: m / z = 225.9 [M+1]. + .

[0184] (2) Synthesis of 9-amino-7-fluoro-1,2,3,4-tetrahydrobenzo[e][1,4]diazepin-5-one (Compound M-29)

[0185] M-28 (2 g, 8.89 mmol) prepared in step (1) was dissolved in MeOH / AcOH (15 mL / 2 mL), and Pd / C (100 mg, 5%) was added to the solution. The reaction was stirred under hydrogen (1 atm) at room temperature for 16 h. After the reaction, the catalyst was filtered off, the reaction solution was concentrated, and the residue was purified by column chromatography using DCM / MeOH (5:1) as the eluent to obtain the product M-29, 450 mg, in a yield of 26%. LCMS of the product M-29: m / z = 195.9 [M+1] + .

[0186] (3) Synthesis of 1-(4-nitrophenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound M-31)

[0187] M-30 (480 mg, 2.88 mmol) was dissolved in thionyl chloride (5 mL) and refluxed at 90°C for 1 h. After the reaction, the reaction solution was concentrated to dryness, and the residue was dissolved in DCM (5 mL). A mixture of the raw material M-29 (510 mg, 2.61 mmol) obtained in step (2) and Py (0.5 mL, 6.21 mmol, density 0.983 g / cm3) in DCM (2 mL) was added and stirred at room temperature for 1 h. After the reaction, the reaction solution was concentrated. TsOH (0.9 g, 5.22 mmol) and MeOH (10 mL) were added to the residue and refluxed at 90°C for 3 h. After the reaction was completed, the reaction solution was cooled, concentrated, dissolved in EA (100 mL), washed with saturated NaHCO3 (2 x 50 mL), and washed with 1N HCl (2 x 50 mL). The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography using EA as the eluent to obtain the product M-31, 418 mg, in a yield of 49%. LCMS of the product M-31: m / z = 326.9 [M+1] + .

[0188] (4) Synthesis of 1-(4-aminophenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound C-11)

[0189] M-31 (418 mg, 1.28 mmol) obtained in step (3) and Pd / C (42 mg, 0.04 mmol, 10%) were dissolved in MeOH (20 mL) and stirred at room temperature under hydrogen (1 atm) for 24 h. After the reaction, the catalyst was filtered off and the mixture was concentrated. The residue was purified by column chromatography using DCM / MeOH (5:1) as the eluent to obtain product C-11 (220 mg, 58%). LCMS of product C-11: m / z = 296.9 [M+1] + .

[0190] Example 22: Synthesis of Intermediate C-12

[0191] Synthesis route of compound C-12:

[0192] (1) Synthesis of 2-fluoro-N-(7-fluoro-5-oxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepin-9-yl)-4-nitrobenzamide (Compound M-33)

[0193] M-29 (500 mg, 2.56 mmol), M-32 (474 ​​mg, 2.56 mmol), HATU (1.46 g, 3.84 mmol), and DIEA (0.67 mL, 3.97 mmol, density 0.766 g / mL) were dissolved in DMF (10 mL) to obtain a mixture. The mixture was stirred at 120°C for 12 h. After the reaction, the reaction solution was cooled and diluted with water (50 mL). The resulting solid was filtered, washed, and dried to obtain the product M-33 (601 mg, 65% yield). LCMS of the product M-33: m / z = 362.8 [M+1] + .

[0194] (2) Synthesis of 1-(4-nitro-2-fluoro-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound M-34)

[0195] M-33 (601 mg, 1.66 mmol) obtained in step (1) was dissolved in AcOH (10 mL) to obtain a mixed solution; the mixed solution was stirred at 110°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and concentrated. The residue was washed with water and DCM, and dried to obtain the product M-34, 516 mg, with a yield of 90%. LCMS of the product M-34: m / z = 344.8 [M+1] + .

[0196] (3) Synthesis of 1-(4-amino-2-fluoro-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (C-12)

[0197] M-34 (516 mg, 1.5 mmol) obtained in step (2) and Pd / C (104 mg, 0.2 mmol, 20%) were dissolved in MeOH / DCM (300 mL / 100 mL). The resulting mixture was stirred under hydrogen (1 atm) at room temperature for 24 h. After the reaction, the catalyst was filtered off and washed with DCM / MeOH (2:1). The organic phase was concentrated to obtain product C-12, 467 mg, with a yield of 99%. LCMS of product C-12: m / z = 314.9 [M+1] + .

[0198] Example 23: Synthesis of Intermediate C-13

[0199] Synthesis route of compound C-13:

[0200] (1) Synthesis of 3-fluoro-N-(7-fluoro-5-oxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepin-9-yl)-4-nitrobenzamide (Compound M-36)

[0201] M-29 (500 mg, 2.56 mmol), M-35 (474 ​​mg, 2.56 mmol), HATU (1.17 g, 3.07 mmol), and DIEA (0.67 mL, 3.97 mmol, density 0.766 g / mL) were dissolved in DMF (10 mL) to obtain a mixture. The mixture was stirred at 120°C for 12 h. After the reaction, the reaction solution was cooled and diluted with water (50 mL). The resulting solid was filtered, washed, and dried to obtain the product M-36, 570 mg, in a yield of 61%. LCMS of the product M-36: m / z = 362.8 [M+1] + .

[0202] (2) Synthesis of 1-(4-nitro-3-fluoro-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound M-37)

[0203] M-36 (570 mg, 1.57 mmol) obtained in step (1) was dissolved in AcOH (10 mL) to obtain a mixed solution; the mixed solution was stirred at 110°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and concentrated. The residue was washed with water and DCM, and dried to obtain the product M-37, 373 mg, 69%. LCMS of the product M-37: m / z = 344.8 [M+1] + .

[0204] (3) Synthesis of 1-(4-amino-3-fluoro-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound C-13)

[0205] M-37 (373 mg, 1.08 mmol) obtained in step (2) and Pd / C (76 mg, 0.14 mmol, 20%) were dissolved in MeOH / DCM (200 mL / 80 mL). The resulting mixture was stirred under hydrogen (1 atm) at room temperature for 24 h. After the reaction, the catalyst was filtered off and washed with DCM / MeOH (2:1). The organic phase was concentrated to obtain product C-13, 336 mg, 99%. LCMS of product C-13: m / z = 314.9 [M+1] + .

[0206] Example 24: Synthesis of Intermediate C-14

[0207] Synthesis route of compound C-14:

[0208] (1) Synthesis of N-(7-fluoro-5-oxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepin-9-yl)-5-nitropyridinecarboxamide (Compound M-39)

[0209] M-29 (500 mg, 2.56 mmol), M-38 (430 mg, 2.56 mmol), HATU (1.17 g, 3.07 mmol), and DIEA (0.67 mL, 3.97 mmol, density 0.766 g / mL) were dissolved in DMF (10 mL) to obtain a mixture. The mixture was stirred at 120°C for 12 h. After the reaction, the reaction solution was cooled and diluted with water (50 mL). The resulting solid was filtered, washed, and dried to obtain the product M-39 (635 mg, 72% yield). LCMS of the product M-39: m / z = 345.8 [M+1] + .

[0210] (2) Synthesis of 1-(5-nitro-pyridin-2-yl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound M-40)

[0211] M-39 (635 mg, 1.84 mmol) prepared in step (1) was dissolved in AcOH (10 mL) to obtain a mixed solution. The mixed solution was stirred at 110°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and concentrated. The residue was washed with water and DCM, and dried to obtain the product M-40, 480 mg, with a yield of 80%. LCMS of the product M-40: m / z = 327.8 [M+1] + .

[0212] (3) Synthesis of 1-(5-amino-pyridin-2-yl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound C-14)

[0213] M-40 (481 mg, 1.47 mmol) obtained in step (2) and Pd / C (48 mg, 0.05 mmol, 10%) were dissolved in MeOH / DCM (200 mL / 80 mL). The resulting mixture was stirred under hydrogen (1 atm) at room temperature for 24 h. After the reaction, the catalyst was filtered off and washed with DCM / MeOH (2:1). The organic phase was concentrated to obtain product C-14, 360 mg, in a yield of 82.4%. LCMS of product C-14: m / z = 297.9 [M+1]+ .

[0214] Example 25: Synthesis of Intermediate C-15

[0215] Synthesis route of compound C-15:

[0216] (1) Synthesis of 4-hydroxy-5-nitronicotinic acid (Compound M-42)

[0217] To a solution of M-41 (24 g, 172.5 mmol) in concentrated sulfuric acid (200 mL) was added concentrated nitric acid (60 mL) dropwise at 0°C for 30 min. The temperature was then slowly raised to room temperature and stirred at 100°C for 20 h. After the reaction, the reaction solution was cooled to room temperature and poured into ice water (1 L) with vigorous stirring. The resulting solid was filtered, washed with water (1 L), and dried to obtain product M-42 (10 g, yield 31.5%). LCMS of product M-42: m / z = 184.9 [M+1] + .

[0218] (2) Synthesis of methyl 4-chloro-5-nitronicotinate (Compound M-43)

[0219] To a mixture of M-42 (10 g, 54.3 mmol, prepared in step (1)) in DCE / DMF (150 mL / 1 mL) was slowly added dichlorothionyl (SOCl2) (28 mL, 386 mmol, density 1.638 g / mL) dropwise to obtain a mixture. The mixture was stirred and refluxed at 84°C for 24 h. After the reaction, the reaction solution was cooled to room temperature and concentrated. The residue was dissolved in DCM (25 mL) and then slowly added dropwise to a mixture of anhydrous methanol / TEA (200 mL / 2 mL) at -10°C. The mixture was stirred for 2 h. After the reaction, the mixture was diluted with water (100 mL) and extracted with DCM (400 mL). The organic phase was washed with brine and dried to obtain the product M-43, 8.51 g, in a yield of 72.5%. LCMS of the product M-43: m / z = 216.9 [M+1]. + .

[0220] (3) Synthesis of 9-nitro-1,2,3,4-tetrahydropyrido[4,3-e][1,4]diazepin-5-one (Compound M-44)

[0221] A solution of M-43 (8 g, 37.0 mmol) prepared in step (2), M-27 (5.38 g, 40.7 mmol), and sodium carbonate (8.63 g, 81.4 mmol) in acetonitrile (150 mL) was stirred at 90°C for 12 h. After the reaction, the reaction solution was cooled to room temperature, concentrated, washed with water, and dried to obtain the product M-44, 3.25 g, with a yield of 42.3%. LCMS of the product M-44: m / z = 208.9 [M+1] + .

[0222] (4) Synthesis of 9-amino-1,2,3,4-tetrahydropyrido[4,3-e][1,4]diazepin-5-one (Compound M-45)

[0223] M-44 (3 g, 14.4 mmol) prepared in step (3) and Pd / C (0.3 g, 0.28 mmol, 10%) were dissolved in MeOH / DCM (300 mL / 150 mL) and stirred at room temperature under hydrogen (1 atm) for 24 h. After the reaction, the catalyst was filtered off and washed with DCM / MeOH (100 mL / 100 mL). The filtrate was concentrated to obtain the product M-45, 2.56 g, in a 99% yield. LCMS of the product M-45: m / z = 179.0 [M+1] + .

[0224] (5) Synthesis of 1-(4-nitrophenyl)-8,9-dihydro-2,4,7,9a-tetraazabenzo[cd]azulen-6(7H)-one (Compound M-46)

[0225] To a solution of M-30 (2.4 g, 14.38 mmol) in DCE (25 mL) was added dropwise thionyl chloride (10 mL, 138 mmol, density 1.638 g / mL) and DMF (1 mL) at room temperature, and the mixture was refluxed at 85°C for 2 h. After the reaction was completed, the reaction solution was cooled to room temperature and concentrated. The residue was dissolved in DCE (50 mL) and added dropwise to a solution of M-45 (2.56 g, 14.38 mmol, prepared in step (4)) and DIEA (2 mL, 11.85 mmol, density 0.766 g / mL) in DCE (100 mL) at room temperature. The mixture was stirred at 90°C for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature and concentrated to dryness. MeOH (100 mL) and MeSO3H (2.76 g, 28.76 mmol) were added to the residue and refluxed at 85°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and concentrated. The residue was purified by column chromatography using EA / MeOH (10:1) as the eluent to obtain product M-46, 286 mg, with a yield of 6.4%. LCMS of product M-46: m / z = 309.7 [M+1] + .

[0226] (6) Synthesis of 1-(4-aminophenyl)-8,9-dihydro-2,4,7,9a-tetraazabenzo[cd]azulen-6(7H)-one (Compound C-15)

[0227] M-46 (286 mg, 0.93 mmol) prepared in step (5) and Pd / C (28.6 mg, 0.03 mmol, 10%) were dissolved in MeOH / DCM (200 mL / 100 mL) to obtain a mixture. The mixture was stirred under hydrogen (1 atm) at room temperature for 12 h. After the reaction, the catalyst was removed by filtration and concentrated to obtain product C-15 (257 mg, 99% yield). LCMS of product C-15: m / z = 279.9 [M+1] + .

[0228] Example 26: Synthesis of Compound I-1

[0229] Synthesis route of compound I-1:

[0230] (1) Synthesis of 2-(4-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound I-1)

[0231] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol, prepared in Example 1), compound C-1 (0.1 g, 0.4 mmol, prepared in Example 11), Pd2dba3 (37 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in dioxane (3 mL) was stirred at 130°C under nitrogen for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 2) to obtain product I-1, 4.6 mg. LCMS of product I-1: m / z=454.8[M+1] + .

[0232] The H NMR spectrum data of product I-1 are: 1 H NMR (400MHz, DMSO-d6): δ=9.20(s,1H),8.64(s,1H),8.12(m,1H),7.95(m,1H),7.90(s,1H),7.80(s,1 H),7.40(m,2H),7.30(m,2H),6.57-6.59(s,1H),5.66(s,2H),2.03(m,1H),0.85(m,2H),0.52(m,2H).

[0233] Example 27: Synthesis of Compound I-2

[0234] Synthesis route of compound I-2:

[0235] (1) Synthesis of 2-(3-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound I-2)

[0236] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol, prepared in Example 1), compound C-2 (0.1 g, 0.4 mmol, prepared in Example 12), Pd2dba3 (38 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 4) to obtain product I-2, 8 mg. LCMS of product I-2: m / z=454.6[M+1] + .

[0237] The H NMR spectrum data of product I-2 are: 1 H NMR (400MHz, DMSO-d6): δ=13.30(s,1H),9.91(s,1H),9.33(s,1H),8.74(s,1H),8.24 (s,1H),8.07(s,1H),7.75-7.87(m,4H),7.19-7.52(m,3H),2.03(s,1H),0.65(s,4H).

[0238] Example 28: Synthesis of Compound I-3

[0239] Synthesis route of compound I-3:

[0240] (1) Synthesis of 2-(4-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2-fluorophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound I-3)

[0241] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol, prepared in Example 1), compound C-3 (0.107 g, 0.4 mmol, prepared in Example 13), Pd2dba3 (38 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in dioxane (3 mL) was stirred at 130°C under nitrogen for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 2) to obtain product I-3, 10 mg. LCMS of product I-3: m / z=471.8[M+1] + .

[0242] The H NMR spectrum data of product I-3 are: 1 H NMR (400MHz, DMSO-d6): δ=12.52-13.00(m,2H),9.29-9.45(s,1H),8.74(s,1H),8.45-8.50(s, 1H),8.02-8.31(s,1H),7.75-7.87(m,3H),7.19-7.52(m,3H),2.03(s,1H),0.83-0.90(m,4H).

[0243] Example 29: Synthesis of Compound I-4

[0244] Synthesis route of compound I-4:

[0245] (1) Synthesis of 2-(5-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2-fluorophenyl)-1H-benzo[d]imidazole-4-carboxamide (Compound I-4)

[0246] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol, prepared in Example 1), compound C-4 (0.107 g, 0.4 mmol, prepared in Example 14), Pd2dba3 (38 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 2) to obtain product I-4, 6.8 mg. LCMS of product I-4: m / z=471.9[M+1] + .

[0247] The H NMR spectrum data of product I-4 are: 1 H NMR (400MHz, DMSO-d6): δ=13.03(s,1H),10.02(s,1H),9.25(s,1H),8.83(s,1H),8 .07-8.20(s,1H),7.75-7.82(m,4H),7.21-7.49(m,3H),2.085(s,1H),0.65(s,4H).

[0248] Example 30: Synthesis of Compound I-5

[0249] Synthesis route of compound I-5:

[0250] (1) Synthesis of 2-(5-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (Compound I-5)

[0251] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol prepared in Example 1), compound C-5 (0.1 g, 0.4 mmol prepared in Example 15), Pd2dba3 (38 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in Dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (DCM / MeOH = 16 / 1) to obtain product I-5, 8 mg. LCMS of product I-5: m / z = 454.8 [M+1] + .

[0252] The H NMR spectrum data of product I-5 are: 1 H NMR (400MHz, DMSO-d6): δ=9.14(s,1H),8.62(s,1H),7.72-8.20(m,8H),7.42-7.49(m,1H),7.01-7.07(m,1H),2.03(m,1H),0.49-0.51(s,4H).

[0253] Example 31: Synthesis of Compound I-6

[0254] Synthesis route of compound I-6:

[0255] (1) Synthesis of 2-(5-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)pyridin-3-yl)-1H-benzo[d]imidazole-4-carboxamide (Compound I-6)

[0256] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol, prepared in Example 1), compound C-6 (0.1 g, 0.4 mmol, prepared in Example 16), Pd2dba3 (38 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (DCM / MeOH = 16 / 1) to obtain product I-6, 6.6 mg. LCMS of product I-6: m / z = 454.8 [M+1] + .

[0257] The H NMR spectrum data of product I-6 are: 1 H NMR (400MHz, DMSO-d6): δ=13.54(s,1H),10.11(s,1H),9.13-9.20(m,3H),8.93(s,1H) ),8.28(s,1H),7.75-7.90(m,3H),7.27-7.40(m,2H),2.03(m,1H),0.52-0.69(s,4H).

[0258] Example 32: Synthesis of Compound I-7

[0259] Synthesis route of compound I-7:

[0260] (1) Synthesis of N-tert-butyl-2-(4-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-2H-indazole-7-carboxamide (Compound M-17)

[0261] A mixed solution of compound A-2-1 (0.1 g, 0.42 mmol, prepared in Example 1), compound C-7 (0.12 g, 0.4 mmol, prepared in Example 17), Pd2dba3 (38 mg, 0.042 mmol), Xantphos (24.3 mg, 0.042 mmol) and Cs2CO3 (0.27 g, 0.84 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 1) to obtain product M-17, 30 mg. LCMS of product M-17: m / z=509.9[M+1] + .

[0262] (2) Synthesis of 2-(4-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-2H-indazole-7-carboxamide (Compound I-7)

[0263] M-17 (0.031 g, 0.06 mmol) prepared in step (1) was dissolved in TFA (1.5 mL) and refluxed at 100°C overnight. After the reaction, the reaction solution was cooled to room temperature, saturated NaHCO3 solution (5 mL) was added, and the mixture was extracted with EA (10 mL*3). The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA / MeOH = 1 / 5 / 0.1) to obtain product I-7, 4.5 mg. LCMS of product I-7: m / z = 453.9 [M+1] + .

[0264] The H NMR spectrum data of product I-7 are: 1 H NMR (400MHz, DMSO-d6): δ=11.62(m,1H),10.00(s,1H),9.23(m,1H),8.58(m,1H),8.10-8.25(m ,3H),8.06-8.16(m,3H),7.89-7.99(m,1H),7.23-7.27(m,2H),2.02(s,1H),0.70-0.90(m,4H).

[0265] Example 33: Synthesis of Compound I-8

[0266] Synthesis route of compound I-8:

[0267] (1) Synthesis of N-tert-butyl-2-(3-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-2H-indazole-7-carboxamide (Compound M-20)

[0268] A mixed solution of compound A-2-1 (0.071 g, 0.3 mmol, prepared in Example 1), compound C-8 (0.092 g, 0.3 mmol, prepared in Example 18), Pd2dba3 (27 mg, 0.03 mmol), Xantphos (34 mg, 0.06 mmol) and Cs2CO3 (0.195 g, 0.6 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA=1 / 1) to obtain product M-20, 38 mg. LCMS of product M-20: m / z=509.9[M+1] + .

[0269] (2) Synthesis of 2-(4-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-2H-indazole-7-carboxamide (Compound I-8)

[0270] M-20 (0.025 g, 0.05 mmol) prepared in step (1) was dissolved in TFA (1.0 mL) and refluxed at 100°C overnight. After the reaction, the reaction solution was cooled to room temperature, saturated NaHCO3 solution (5 mL) was added, and the mixture was extracted with EA (10 mL*3). The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA / MeOH = 1 / 5 / 0.1) to obtain product I-8, 9 mg. LCMS of product I-8: m / z = 453.8 [M+1] + .

[0271] The H NMR spectrum data of product I-8 are: 1 H NMR (400MHz, DMSO-d6): δ=9.88(s,1H),8.37-8.45(m,2H),8.16-8.23(d,1H),7.72-7.93(m1H),7.65-7.70 (m,2H),7.50-7.52(s,1H),7.23-7.35(m,3H),7.10-7.20(m,2H),2.02(s,1H),0.43(s,2H),0.185(s,2H).

[0272] Example 34: Synthesis of Compound I-9

[0273] Synthesis route of compound I-9:

[0274] (1) Synthesis of 2-(4-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-8-fluoro-4,5-dihydro-1H-aza[5,4,3-cd]indol-6(3H)-one (Compound I-9)

[0275] A mixed solution of compound A-2-1 (0.065 g, 0.271 mmol, prepared in Example 1), compound C-9 (0.08 g, 0.271 mmol, prepared in Example 19), Pd2dba3 (50 mg, 0.054 mmol), Xantphos (62 mg, 0.108 mmol) and Cs2CO3 (0.176 g, 0.542 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 12 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA / MeOH = 1 / 4 / 0.1) to obtain product I-9, 25 mg. LCMS of product I-9: m / z = 496.8 [M+1] + .

[0276] The H NMR spectrum data of product I-9 are: 1 H NMR (400MHz, DMSO-d6): δ = 11.56 (s, 1H); 9.89 (s, 1H), 8.21-8.24 (m, 2H), 8.07-8.10 (m, 2H), 7.56-7.58 (m, 2H), 7.39-7. 42(m,1H),7.30-7.35(m,1H),7.28(s,1H),3.39(m,2H),3.04(m,2H),2.88(m,1H),0.85-0.86(m,2H),0.76-0.79(m,2H).

[0277] Example 35: Synthesis of Compound I-10

[0278] Synthesis route of compound I-10:

[0279] (1) Synthesis of 2-(3-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-8-fluoro-4,5-dihydro-1H-aza[5,4,3-cd]indol-6(3H)-one (Compound I-10)

[0280] A mixed solution of compound A-2-1 (0.015 g, 0.06 mmol, prepared in Example 1), compound C-10 (0.018 g, 0.06 mmol, prepared in Example 20), Pd2dba3 (6 mg, 0.006 mmol), Xantphos (6 mg, 0.012 mmol), and Cs2CO3 (0.059 g, 0.18 mmol) in dioxane (3 mL) was stirred under nitrogen at 130°C for 4 hours. After the reaction, the reaction solution was cooled, water (6 mL) was added, and EA (10 mL*3) was extracted. The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (PE / EA / MeOH = 1 / 5 / 0.1) to obtain product I-10, 6 mg. LCMS of product I-10: m / z = 496.9 [M+1] + .

[0281] The H NMR spectrum data of product I-10 are: 1 H NMR (400MHz, DMSO-d6): δ=11.60(s,1H),9.84(s,1H),8.23(s,1H),8.14(s,1H),7.99-8.02(s,1H),7.11 -7.45(m,6H),3.31-3.39(d,2H),2.85-3.03(d,2H),2.02(m,1H),0.83-0.87(m,2H),0.58-0.61(m,2H).

[0282] Example 36: Synthesis of Compound I-11

[0283] Synthesis route of compound I-11:

[0284] (1) Synthesis of 1-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-11)

[0285] A mixture of compound A-2-1 (50 mg, 0.21 mmol, prepared in Example 1), C-11 (63 mg, 0.21 mmol, prepared in Example 21), Pd2dba3 (19 mg, 0.021 mmol), Xantphos (12 mg, 0.021 mmol), and Cs2CO3 (137 mg, 0.42 mmol) in dioxane (3 mL) was stirred at 130°C under nitrogen for 12 hours. After completion of the reaction, the reaction solution was cooled, added with water (6 mL), and extracted with EA (10 mL*3). The organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography using EA / MeOH (20:1) as the eluent to obtain product I-11, 8.2 mg, in a yield of 7.9%. LCMS of product I-11: m / z = 497.8 [M+1]. + .

[0286] The H NMR spectrum data of product I-11 are: 1 H NMR (400MHz, DMSO-d6): δ=10.05(s,1H);8.61(s,1H),8.26(s,1H),8.15-8.16(m,2H),7.81-7.83(m,2H),7.71- 7.73(m,1H),7.56-7.58(m,1H),7.29(s,1H),4.49(m,2H),3.55(m,2H),2.89(m,1H),0.86(m,2H),0.71(m,2H).

[0287] Example 37: Synthesis of Compound I-12

[0288] Synthesis route of compound I-12:

[0289] (1) Synthesis of 1-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-2-fluoro-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-12)

[0290] Compound A-2-1 (99 mg, 0.42 mmol, prepared in Example 1), C-12 (120 mg, 0.38 mmol, prepared in Example 22), and MeSO₃H (37 mg, 0.38 mmol) were dissolved in DMF (2 mL) to obtain a mixture. The mixture was stirred at 120°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and saturated sodium bicarbonate (20 mL) was added to form a solid. The solid was filtered, washed with water, and then washed with DCM. The residue was purified by column chromatography using EA / MeOH (20:1) as the eluent to obtain product I-12 (18.2 mg, 8.4% yield). LCMS of product I-12: m / z = 515.8 [M+1] + .

[0291] The H NMR spectrum data of product I-12 are: 1 H NMR (400MHz, DMSO-d6): δ = 10.28 (s, 1H), 8.62 (s, 1H), 8.4 (d, 1H), 7.73-7.75 (m, 2H), 7.64-7.66 ( m,2H),7.64(s,1H),4.26(s,2H),3.56(s,2H),2.85(s,1H),0.85-0.86(m,3H),0.72-0.85(s,2H).

[0292] Example 38: Synthesis of Compound I-13

[0293] Synthesis route of compound I-13:

[0294] (1) Synthesis of 1-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-3-fluoro-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-13)

[0295] A-2-1 (99 mg, 0.42 mmol, prepared in Example 1), C-13 (120 mg, 0.38 mmol, prepared in Example 23), and MeSO₃H (37 mg, 0.38 mmol) were dissolved in DMF (2 mL) to obtain a mixture. The mixture was stirred at 120°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and saturated sodium bicarbonate (20 mL) was added to form a solid. The solid was filtered, washed with water, and then washed with DCM. The residue was purified by column chromatography using EA / MeOH (20:1) as the eluent to obtain the product I-13, 12 mg, in a yield of 6.1%. LCMS of the product I-13: m / z = 515.8 [M+1]. + .

[0296] The H NMR spectrum data of product I-13 are: 1 H NMR (400MHz, DMSO-d6): δ=9.39(s,1H),8.62(m,1H),8.42(m,1H),8.24(s,1H),7.69-7.76(m,3H), 7.58-7.61(d,j=12Hz,1H),7.24(s,1H),4.5(m,2H),3.56(m,2H),2.82(m,1H),0.74-0.86(m,4H).

[0297] Example 39: Synthesis of Compound I-14

[0298] Synthesis route of compound I-14:

[0299] (1) Synthesis of 1-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-2-pyridyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-14)

[0300] Compound A-2-1 (105 mg, 0.44 mmol, prepared in Example 1), C-14 (120 mg, 0.40 mmol, prepared in Example 24), and MeSO₃H (39 mg, 0.40 mmol) were dissolved in DMF (2 mL) to obtain a mixture. The mixture was stirred at 120°C for 12 h. After the reaction, the reaction solution was cooled to room temperature and saturated sodium bicarbonate (20 mL) was added to form a solid. The solid was filtered, washed with water, and then washed with DCM. The residue was purified by column chromatography using EA / MeOH (30:1) as the eluent to obtain product I-14 (65 mg, 32.6% yield). LCMS of product I-14: m / z = 498.8 [M+1] + .

[0301] The H NMR spectrum data of product I-14 are: 1 H NMR (400MHz, DMSO-d6): δ=10.25(s,1H),9.13(s,1H),8.70-8.73(s,1H),8.59(s,1H),8.27-8.29(s,2H),7.73-7.75(d ,j=8Hz,1H),7.61-7.64(d,j=12Hz,1H),7.37(s,1H),3.26(m,2H),2.89(m,2H),1.99(m,1H),0.85(m,2H),0.72(m,2H).

[0302] Example 40: Synthesis of Compound I-15

[0303] Synthesis route of compound I-15:

[0304] (1) Synthesis of 1-(4-(cyclopropylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-8,9-dihydro-2,4,7,9a-tetraazabenzo[cd]azulen-6-one (Compound I-15)

[0305] Compound A-2-1 (140 mg, 0.59 mmol, prepared in Example 1), C-15 (150 mg, 0.54 mmol, prepared in Example 25), and methanesulfonic acid (52 mg, 0.54 mmol) were dissolved in DMF (2.5 mL) to obtain a mixture. The mixture was stirred at 120°C for 6 h. After the reaction, the reaction solution was cooled to room temperature, saturated sodium bicarbonate aqueous solution was added, and stirred for 2 h. The precipitated solid was filtered, washed with water, washed with DCM, and dried to obtain product I-15, 84.7 mg, with a yield of 32.7%. LCMS of product I-15: m / z = 480.9 [M+1] + .

[0306] The H NMR spectrum data of product I-15 are: 1 H NMR (400MHz, DMSO-d6): δ = 10.08 (s, 1H); 9.07 (s, 1H), 8.81 (s, 1H), 8.59-8.60 (m, 1H), 8.27 (s, 1H), 8.17-8.19 ( m,2H),7.85-7.87(m,2H),7.31(s,1H),4.51(m,2H),3.58(m,2H),2.89(m,1H),0.86-0.88(m,2H),0.71(m,2H).

[0307] Example 41: Synthesis of Compound I-16

[0308] Synthesis route of compound I-16:

[0309] (1) Synthesis of 5-(4-(4-(cyclobutylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-8-fluoro-3,4-dihydro-2H-aza[5,4,3-cd]indol-1(6H)-one (Compound I-16)

[0310] Compound A-2-2 (0.102 g, 0.407 mmol, prepared in Example 2), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), TsOH (0.06 g, 0.334 mmol) and Dioxane (2 mL) were mixed and refluxed at 80°C for 12 h. After the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20*3 mL), dried, and concentrated. PE / EA=25 / 1 was used to obtain product I-16, 0.06 g. LCMS of product I-16: m / z=510.8 (M+1) + .

[0311] The H NMR spectrum data of product I-16 are: 1 H NMR (400MHz, DMSO-d6): δ=11.50(s,1H),9.92(s,1H),8.22-8.24(m,2H),7.91-7.93(d,2H),7.51-7.59(d,2 H),7.40(m,1H),7.3(m,1H),7.06(m,1H),4.7(m,1H),3.5(m,2H),3.1(m,2H),2.2-2.4(m,4H),1.73(m,2H).

[0312] Example 42: Synthesis of Compound I-17

[0313] Synthesis route of compound I-17:

[0314] (1) Synthesis of 8-fluoro-5-(4-(4-(oxetane-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-3,4-dihydro-2H-azacyclo[5,4,3-cd]indol-1(6H)-one (Compound I-17)

[0315] Compound A-2-3 (0.103 g, 0.407 mmol, prepared in Example 3), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), TsOH (0.06 g, 0.334 mmol) and Dioxane (2 mL) were mixed and refluxed at 80°C for 12 h. After the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20*3 mL), dried, and concentrated. PE / EA=25 / 1 was used to obtain product I-17, 0.013 g. LCMS of product I-17: m / z=512.8 (M+1) + .

[0316] The H NMR spectrum data of product I-17 are:1 H NMR (400MHz, DMSO-d6): δ=11.68(s,1H),9.83(s,1H),8.42(s,1H),8.29(s,1H),7.83-7.85 (d,2H),7.60-7.66(s,3H),7.25-7.33(m,2H),4.69-4.82(m,4H),3.33(s,3H),3.05(s,2H).

[0317] Example 43: Synthesis of Compound I-18

[0318] Synthesis route of compound I-18:

[0319] (1) Synthesis of 5-(4-(4-((1r,4r)-4-aminocyclohexylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-8-fluoro-3,4-dihydro-2H-azepinyl[5,4,3-cd]indol-1(6H)-one (Compound I-18)

[0320] Compound A-2-5 (0.157 g, 0.407 mmol, prepared in Example 4), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), Pd2dba3 (0.031 g, 0.0334 mmol), Xantphos (20 mg, 0.034 mmol) and Cs2CO3 (326 mg, 1 mmol) were dissolved in dioxane (2 mL) and refluxed at 130°C for 12 h under nitrogen protection. After the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20*3 mL), dried, and concentrated. PE / EA = 1 / 4 was used to obtain 0.03 g of the intermediate. HCl / dioxane (4 M, 2 mL) was added to the obtained intermediate and the reaction was stirred at room temperature overnight. After the reaction, the reaction solution was concentrated to obtain the product I-18, 4.8 mg. LCMS: m / z = 553.5 (M+1) + .

[0321] Example 44: Synthesis of Compound I-19

[0322] Synthesis route of compound I-19:

[0323] (1) Synthesis of 8-fluoro-5-(4-(4-(tetrahydrofuran-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-3,4-dihydro-2H-aza[5,4,3-cd]indol-1(6H)-one (Compound I-19)

[0324] Compound A-2-6 (0.107 g, 0.407 mmol, prepared in Example 5), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), Pd2dba3 (0.031 g, 0.0334 mmol), Xantphos (20 mg, 0.034 mmol) and Cs2CO3 (326 mg, 1 mmol) were dissolved in dioxane (2 mL) and refluxed at 130°C for 12 h. After the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20*3 mL), dried, and concentrated. The PE / EA ratio was 1 / 4 to obtain product I-19, 0.025 g. LCMS: m / z=527.8 (M+1) + .

[0325] The H NMR spectrum data of product I-19 are: 1 H NMR (400MHz, DMSO-d6): δ=11.65(s,1H),9.85(s,1H),8.15-8.26(m,2H),7.88-7.90(d,2H), 7.7.57-7.59(d,2H),7.25-7.42(m,3H),3.34-3.97(m,6H),3.04(s,3H),2.16-2.24(m,2H).

[0326] Example 45: Synthesis of Compound I-20

[0327] Synthesis route of compound I-20:

[0328] (1) Synthesis of 8-fluoro-5-(4-(4-(tetrahydro-2H-pyran-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-3,4-dihydro-2H-azapyridinyl[5,4,3-cd]indol-1(6H)-one (Compound I-20)

[0329] Compound A-2-7 (0.112 g, 0.407 mmol, prepared in Example 6) and C-9 (0.1 g, 0.334 mmol, prepared in Example 19) were dissolved in HCl / Dioxane (4 M, 2 mL) and reacted at 80°C in a sealed tube for 12 h. After the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, the pH was adjusted to 7-8, and the solution was extracted with ethyl acetate (20*3 mL), dried, and concentrated. The PE / EA ratio was 1 / 4 to obtain product I-20, 0.07 g. LCMS: m / z=540.8 (M+1) + .

[0330] The H NMR spectrum data of product I-20 are: 1 H NMR (400MHz, DMSO-d6): δ=11.37(s,1H),9.85(s,1H),8.16-8.25(m,2H),7.56-7.58(d,2H) ,7.22-7.39(m,5H),3.94(d,2H),3.33-3.50(s,4H),2.90-3.10(m,3H),1.81-1.80(m,4H).

[0331] Example 46: Synthesis of Compound I-21

[0332] Synthesis route of compound I-21:

[0333] (1) Synthesis of 8-fluoro-5-(4-(4-(piperidin-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-3,4-dihydro-2H-aza[5,4,3-cd]indol-1(6H)-one (Compound I-21)

[0334] Compound A-2-8 (0.152 g, 0.407 mmol, Example 7), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), Pd2dba3 (0.031 g, 0.0334 mmol), Xantphos (20 mg, 0.034 mmol), and Cs2CO3 (326 mg, 1 mmol) were dissolved in dioxane (2 mL) and refluxed at 130°C for 12 h under nitrogen. After completion of the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, the pH was adjusted to 7-8, and the solution was extracted with ethyl acetate (20 x 3 mL), dried, and concentrated to a PE / EA ratio of 1 / 4 to obtain 0.06 g of the intermediate. The obtained intermediate was added with HCl / dioxane (4 M, 2 mL) and stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated to obtain 60 mg of the product I-21. LCMS: m / z=540.8 (M+1) + .

[0335] The H NMR spectrum data of product I-21 are: 1H NMR (400MHz, DMSO-d6): δ=11.78(s,1H),9.00(s,1H),8.26(s,2H),8.33(s,1H),7.78-7.86(d,2H ),7.62-7.64(d,2H),7.26-7.36(m,2H),3.30-3.45(m,4H),2.98-3.05(m,5H),1.83-2.02(d,5H).

[0336] Example 47: Synthesis of Compound I-22

[0337] Synthesis route of compound I-22:

[0338] (1) Synthesis of 8-fluoro-5-(4-(4-(pyrrolidin-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-3,4-dihydro-2H-aza[5,4,3-cd]indol-1(6H)-one (Compound I-22)

[0339] Compound A-2-9 (0.146 g, 0.407 mmol, prepared in Example 8), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), Pd2dba3 (0.031 g, 0.0334 mmol), Xantphos (20 mg, 0.034 mmol), and Cs2CO3 (326 mg, 1 mmol) were dissolved in dioxane (2 mL) and refluxed at 130°C for 12 h under nitrogen. After completion of the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, the pH was adjusted to 7-8, and the solution was extracted with ethyl acetate (20 x 3 mL), dried, and concentrated to a PE / EA ratio of 1 / 4 to obtain 0.048 g of the intermediate. The obtained intermediate was added with HCl / dioxane (4 M, 2 mL) and stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated to obtain 30 mg of the product I-22. LCMS: m / z=526.8 (M+1) + .

[0340] The H NMR spectrum data of product I-22 are: 1 H NMR (400MHz, DMSO-d6): δ=11.85(s,1H),9.83(s,1H),8.21-8.24(s,2H),7.88-7.98(s,2H),7 .53-7.58(d,2H),7.26-7.40(m,3H),3.45-3.72(m,4H),2.78-3.09(m,5H),2.01-2.08(d,3H).

[0341] Example 48: Synthesis of Compound I-23

[0342] Synthesis route of compound I-23:

[0343] (1) Synthesis of 5-(4-(4-((1S,3S)-3-aminocyclopentylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-8-fluoro-3,4-dihydro-2H-azepinyl[5,4,3-cd]indol-1(6H)-one (Compound I-23)

[0344] Compound A-2-10 (0.152 g, 0.407 mmol, prepared in Example 9), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), Pd2dba3 (0.031 g, 0.0334 mmol), Xantphos (20 mg, 0.034 mmol), and Cs2CO3 (326 mg, 1 mmol) were dissolved in dioxane (2 mL) and refluxed at 130°C for 12 h under nitrogen. After completion of the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, the pH was adjusted to 7-8, and the solution was extracted with ethyl acetate (20 x 3 mL), dried, and concentrated to a PE / EA ratio of 1 / 4 to obtain 0.07 g of the intermediate. The obtained intermediate was added with HCl / dioxane (4 M, 2 mL) and stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated to obtain the product I-23 (35 mg). LCMS: m / z=539.9 (M+1) + .

[0345] The H NMR spectrum data of product I-23 are: 1 H NMR (400MHz, DMSO-d6): δ=11.62(s,1H),9.81(s,1H),8.19-8.21(s,2H),7.93-7.95(s,2H),7.55-7.7(d,2H) ,7.26-7.42(m,3H),3.47(m,2H),2.98-3.15(m,4H),2.08-2.20(m,2H),1.85-2.02(m,2H),1.45-1.82(m,4H).

[0346] Example 49: Synthesis of Compound I-24

[0347] Synthesis route of compound I-24:

[0348] (1) Synthesis of 8-fluoro-5-(4-(4-(3-(2-oxopyrrolidin-1-yl)propylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phenyl)-3,4-dihydro-2H-azepane[5,4,3-cd]indol-1(6H)-one (Compound I-24)

[0349] Compound A-2-11 (0.129 g, 0.407 mmol, prepared in Example 10), C-9 (0.1 g, 0.334 mmol, prepared in Example 19), TsOH (0.06 g, 0.334 mmol) and dioxane (3 mL) were mixed and refluxed at 80°C for 12 h. After the reaction, the reaction solution was cooled to room temperature, 10 mL of water was added, the pH was adjusted to 7-8, and the solution was extracted with ethyl acetate (20*3 mL), dried, concentrated, and then plated with DCM / EA = 1 / 3, and concentrated to obtain product I-24, 20 mg. LCMS: m / z = 582.0 (M+1) + .

[0350] The H NMR spectrum data of product I-24 are: 1 H NMR (400MHz, DMSO-d6): δ=11.62(s,1H),9.82(s,1H),8.23(s,2H),7.90-7.92(d,2H),7.51-7.59(d,2H),7.22-7.34 (m,2H),6.67(s,1H),3.32-3.45(m,4H),2.85-3.15(m,2H),2.15-2.25(m,2H),1.56-2.05(m,4H),1.1-1.32(m,4H).

[0351] Example 50: Synthesis of Compound I-25

[0352] Synthesis route of compound I-25:

[0353] (1) Synthesis of 1-(4-(cyclobutylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-25)

[0354] Compound A-2-2 (30 mg, 0.12 mmol, prepared in Example 2), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, 5 mL of water and 5 mL of ethyl acetate were added, the pH was adjusted to 7-8, and the solution was filtered. The filtrate was separated, and the filter cake and the organic phase were combined, concentrated, and washed with DCM / MeOH = 10 / 1 to obtain the product I-25, 7 mg. LCMS: m / z = 511.8 (M+1) + .

[0355] The H NMR spectrum data of product I-25 are: 1 H NMR (400MHz, DMSO-d6): δ=9.96(s,1H),8.62(s,1H),8.27(s,1H),8.01(d,2H),7.74-7.81(d,2H),7.71-7.74(m,1H) ,7.56-7.59(d,1H),7.12(s,1H),4.49(s,2H),3.56(s,2H),3.01-3.21(m,1H),2.01-2.45(m,4H),1.70-1.73(m,2H).

[0356] Example 51: Synthesis of Compound I-26

[0357] Synthesis route of compound I-26:

[0358] (1) Synthesis of 1-(4-(oxetane-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-26)

[0359] Compound A-2-3 (30 mg, 0.12 mmol, prepared in Example 3), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, 5 mL of water and 5 mL of ethyl acetate were added, the pH was adjusted to 7-8, and the solution was filtered. The filtrate was separated, and the filter cake and the organic phase were combined, concentrated, and washed with DCM / MeOH = 10 / 1 to obtain the product I-26, 8 mg. LCMS: m / z = 513.8 (M+1) + .

[0360] The H NMR spectrum data of product I-26 are:1 H NMR (400MHz, DMSO-d6): δ=9.45(s,1H),8.62(s,1H),8.12(s,2H),7.92-8.1(m,4H),7.62- 7.83(m,2H),4.35-4.45(m,2H),4.02-4.18(d,2H),3.88-3.92(d,2H),3.52-3.68(m,3H).

[0361] Example 52: Synthesis of Compound I-27

[0362] Synthesis route of compound I-27:

[0363] (1) Synthesis of 1-(4-(4-((1r,4r)-4-aminocyclohexylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-27)

[0364] Compound A-2-5 (50 mg, 0.12 mmol, prepared in Example 4), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted at 100°C in a sealed tube overnight. After the reaction, the reaction solution was cooled to room temperature, concentrated, added with 5 mL of petroleum ether / ethyl acetate (1 / 1), slurried, filtered, and the filter cake was added with 2 mL of TFA / DCM (1 / 1). The reaction was allowed to react at room temperature for 4 h. After the reaction, the reaction solution was concentrated, added with 5 mL of water and 5 mL of ethyl acetate, and the pH was adjusted to 7-8. The solution was filtered, the filtrate was separated, the filter cake and the organic phase were combined, concentrated, and plated with DCM / MeOH = 10 / 1 to obtain product I-27, 5 mg. LCMS: m / z = 554.8 (M+1) + .

[0365] The H NMR spectrum data of product I-27 are: 1 H NMR (400MHz, DMSO-d6): δ=10.04(s,1H),8.62(s,1H),8.26(s,1H),8.01-8.21(m,3H),7.81-7.84(d,2H),7.73-7.74(d,1H) ,7.712-7.718(d,1H),4.49(s,2H),3.56(s,2H),2.9-3.01(m,1H),2.56-2.62(m,1H),1.98-2.15(m,5H),1.45-1.62(m,5H).

[0366] Example 53: Synthesis of Compound I-28

[0367] Synthesis route of compound I-28:

[0368] (1) Synthesis of 1-(4-(4-(tetrahydrofuran-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-28)

[0369] Compound A-2-6 (32 mg, 0.12 mmol, prepared in Example 5), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, 5 mL of water and 5 mL of ethyl acetate were added, the pH was adjusted to 7-8, and the solution was filtered. The filtrate was separated, and the filter cake and the organic phase were combined, concentrated, and washed with DCM / MeOH = 10 / 1 to obtain the product I-28, 8 mg. LCMS: m / z = 527.8 (M+1) + .

[0370] The H NMR spectrum data of product I-28 are: 1 H NMR (400MHz, DMSO-d6): δ=10.01(s,1H),8.61(s,1H),8.29(s,1H),7.97-7.99(d,2H),7.81-7.84(d,2H),7.71 -7.713(d,1H),7.756-7.58(d,1H),6.88-6.90(s,1H),4.48-4.49(s,2H),3.56-4.1(m,5H),1.98-2.25(m,4H).

[0371] Example 54: Synthesis of Compound I-29

[0372] Synthesis route of compound I-29:

[0373] (1) Synthesis of 1-(4-(4-(tetrahydro-2H-pyran-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-29)

[0374] Compound A-2-7 (31 mg, 0.12 mmol, prepared in Example 6), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, 5 mL of water and 5 mL of ethyl acetate were added, the pH was adjusted to 7-8, and the solution was filtered. The filtrate was separated, and the filter cake and the organic phase were combined, concentrated, and washed with DCM / MeOH = 10 / 1 to obtain the product I-29, 6 mg. LCMS: m / z = 542.8 (M+1) + .

[0375] The H NMR spectrum data of product I-29 are: 1 H NMR (400MHz, DMSO-d6): δ=10.10(s,1H),8.69(s,1H),8.29(s,1H),7.98-8.02(d,2H),7.82-7.85(d,2H),7.79-7.796(d,1H) ),7.61-7.64(m,1H),6.87-6.90(s,1H),4.51(s,2H),4.35(s,1H),3.93-3.95(d,2H),3.30-3.52(m,4H),1.75-1.80(s,4H).

[0376] Example 55: Synthesis of Compound I-30

[0377] Synthesis route of compound I-30:

[0378] (1) Synthesis of 1-(4-(4-(piperidin-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-30)

[0379] Compound A-2-8 (46 mg, 0.12 mmol, prepared in Example 7), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, concentrated, and added with 5 mL of petroleum ether / ethyl acetate (1 / 1). The mixture was slurried and filtered. The filter cake was added with 2 mL of TFA / DCM (1 / 1) and reacted at room temperature for 4 h. After the reaction, the reaction solution was concentrated, and 5 mL of water and 5 mL of ethyl acetate were added. The pH was adjusted to 7-8, filtered, and the filtrate was separated. The filter cake and the organic phase were combined, concentrated, and plated with DCM / MeOH = 10 / 1 to obtain product I-30, 5 mg. LCMS: m / z = 540.9 (M+1) + .

[0380] The H NMR spectrum data of product I-30 are: 1 H NMR (400MHz, DMSO-d6): δ=10.07(s,1H),8.95(s,1H),8.64(s,1H),8.30(s,1H),7.98-8.02(d,2H),7.82-7.85(d,2H),7.79-7.796(d,1H),7. 61-7.64(m,1H),6.87-6.90(s,1H),4.51(s,2H),4.35(s,1H),3.52-3. 70(s,2H),3.28-3.35(s,1H),2.95-3.12(s,3H),1.1.82-2.03(m,4H).

[0381] Example 56: Synthesis of Compound I-31

[0382] Synthesis route of compound I-31:

[0383] (1) Synthesis of 1-(4-(4-(pyrrolidin-3-ylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-31)

[0384] Compound A-2-9 (44 mg, 0.12 mmol, prepared in Example 8), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, concentrated, and added with 5 mL of petroleum ether / ethyl acetate (1 / 1). The mixture was slurried and filtered. The filter cake was added with 2 mL of TFA / DCM (1 / 1) and reacted at room temperature for 4 h. After the reaction, the reaction solution was concentrated, and 5 mL of water and 5 mL of ethyl acetate were added. The pH was adjusted to 7-8, filtered, and the filtrate was separated. The filter cake and the organic phase were combined, concentrated, and plated with DCM / MeOH = 9 / 1 to obtain product I-31, 10 mg. LCMS: m / z = 526.8 (M+1) + .

[0385] The H NMR spectrum data of product I-31 are: 1 H NMR (400MHz, DMSO-d6): δ=10.11(s,1H),8.65(s,1H),8.32(s,1H),7.96-8.02(d,2H),7.85-7.87(d,2H),7 .79-7.796(d,1H),7.61-7.64(d,1H),7.19(s,1H),3.45-3.72(m,4H),278-3.09(m,5H),2.01-2.08(d,3H).

[0386] Example 57: Synthesis of Compound I-32

[0387] Synthesis route of compound I-32:

[0388] (1) Synthesis of 1-(4-(4-((1S,3S)-3-aminocyclopentylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-32)

[0389] Compound A-2-10 (46 mg, 0.12 mmol, prepared in Example 9), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, concentrated, and added with 5 mL of petroleum ether / ethyl acetate (1 / 1). The mixture was slurried and filtered. The filter cake was added with 2 mL of TFA / DCM (1 / 1) and reacted at room temperature for 4 h. After the reaction, the reaction solution was concentrated, and 5 mL of water and 5 mL of ethyl acetate were added. The pH was adjusted to 7-8, filtered, and the filtrate was separated. The filter cake and the organic phase were combined, concentrated, and plated with DCM / MeOH = 9 / 1 to obtain the product I-32, 7 mg. LCMS: m / z = 540.8 (M+1) + .

[0390] The H NMR spectrum data of product I-32 are: 1 H NMR (400MHz, DMSO-d6): δ=10.07(s,1H),8.65(s,1H),8.27(s,1H),7.96-8.02(d,2H),7.85-7.87(d,2H),7.79-7.796(d,1H) ),7.61-7.64(d,1H),6.95(s,1H),3.47(m,2H),2.98-3.15(m,4H),2.08-2.20(m,2H),1.85-2.02(m,2H),1.45-1.82(m,4H).

[0391] Example 58: Synthesis of Compound I-33

[0392] Synthesis route of compound I-33:

[0393] (1) Synthesis of 1-(4-(4-(3-(2-oxopyrrolidin-1-yl)propylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-4-fluoro-8,9-dihydro-7H-2,7,9a-triaza-benzo[cd]azulen-6-one (Compound I-33)

[0394] Compound A-2-11 (39 mg, 0.12 mmol, prepared in Example 10), C-11 (30 mg, 0.1 mmol, prepared in Example 21), TFA (12 mg, 0.1 mmol) and IPA (2 mL) were mixed and reacted overnight at 100°C in a sealed tube. After the reaction, the reaction solution was cooled to room temperature, 5 mL of water and 5 mL of ethyl acetate were added, the pH was adjusted to 7-8, and the solution was filtered. The filtrate was separated, and the filter cake and the organic phase were combined, concentrated, and washed with DCM / MeOH = 10 / 1 to obtain the product I-33, 12 mg. LCMS: m / z = 582.8 (M+1) + .

[0395] The H NMR spectrum data of product I-33 are: 1 H NMR (400MHz, DMSO-d6): δ = 9.98 (s, 1H); 8.60-8.61 (m, 1H), 8.26 (s, 1H), 7.97-8.00 (m, 2H), 7.82-7.84 (m, 2H), 7.72-7.74 (m, 1H), 7.56- 7.59(m,1H),7.28(m,1H),4.50(m,2H),3.56(m,2H),3.46(m,2H),3.25-3.32(m,4H),2.50(m,2H),1.90-1.92(m,2H),1.78-1.81(m,2H).

[0396] Example 59: Synthesis of Compound I-34

[0397] Synthesis route of compound I-34:

[0398] (1) Synthesis of 1-(4-(4-(3-(2-oxopyrrolidin-1-yl)propylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino-phenyl)-8,9-dihydro-2,4,7,9a-tetraazabenzo[cd]azulen-6-one (Compound I-34)

[0399] Compound A-2-11 (254 mg, 0.79 mmol, prepared in Example 10), C-15 (201 mg, 0.72 mmol, prepared in Example 25), and methanesulfonic acid (69 mg, 0.72 mmol) were dissolved in DMF (2.5 mL) to obtain a mixture. The mixture was stirred at 120°C for 6 h. After the reaction, the reaction solution was cooled to room temperature, saturated sodium bicarbonate aqueous solution was added, and stirred for 2 h. The precipitated solid was filtered, washed with water, washed with DCM, and dried to obtain product I-34, 85.8 mg, in a yield of 21%. LCMS: m / z = 565.8 (M+1). + .

[0400] The H NMR spectrum data of product I-34 are: 1 H NMR (400MHz, DMSO-d6): δ = 10.01 (s, 1H); 9.08 (s, 1H), 8.81 (s, 1H), 8.59-8, 60 (m, 1H), 8.26 (s, 1H), 7.98-8.02 (m, 2H), 7.86-7.89 ( m,2H),7.28(m,1H),4.51(m,2H),3.58(m,2H),3.48(m,2H),3.20-3.30(m,4H),2.20(m,2H),1.90-1.92(m,2H),1.78-1.81(m,2H).

[0401] Example 60: Determination of the in vitro inhibitory activity (IC50 value) of different compounds against Parp1 and ULK1

[0402] 1. Determination of the in vitro inhibitory activity (IC50 value) of different compounds against Parp1

[0403] PARP1 enzyme activity transfers ADP-ribose from the NAD+ substrate to the PAR chain of the histone substrate. The amount of NAD+ remaining after the enzymatic reaction was determined using Promega NAD / NADH-Glo TM The detection kit was used to quantitatively measure PARP1 activity as the endpoint.

[0404] 1.1. Reagents: Reaction buffer: 50 mM Tris-HCl (pH 8.0), 50 mM NaCl, 10 mM MgCl2, 0.01% Brij35, 1 mM DTT, 1% DMSO and 10 mg / mL activated DNA (Sigma cat#D4522).

[0405] 1.2. Substrate: chicken core histone, NAD.

[0406] 1.3. Standard reaction conditions:

[0407] PARP1: 2.5nM (RBC CAT#PAR-21-346);

[0408] Chicken core histone: 0.01 mg / mL (RBC CAT#HMT-35-435);

[0409] NAD: 0.5mM (Sigma-Aldrich CAT#N1636);

[0410] Detection Kit: NAD / NADH-Glo TM(Promega CAT#G9071).

[0411] 1.4 Reaction steps:

[0412] (i) Add 10 mL of 1X enzyme / histone substrate mixture prepared in reaction buffer to the wells of the reaction plate (Corning 3572, untreated). For the total NAD+ signal wells (no enzyme), add histone substrate prepared in reaction buffer.

[0413] (ii) Compounds (prepared in Examples 26-59, respectively) dissolved in 100% DMSO were added to the enzyme mixture using acoustic technology (Echo 550; nanoliter range). 100% DMSO was added to the no-compound / DMSO control wells. The reaction plate was spun down to mix and pre-incubated at room temperature for 20 minutes.

[0414] (iii) Using a Mantis liquid handler, add 100 nL of 100X NAD-stimulated reaction. Spin down the reaction plate to mix. Seal the reaction plate and incubate at room temperature for 2 hours.

[0415] (iv) Add 10 mL of NAD / NADH-Glo prepared according to the test kit instructions. TM Add to all assay wells. Spin down the plate to mix. Seal the plate and incubate in the dark at room temperature for 30 minutes (with black lid).

[0416] (v) Read the endpoint fluorescence value at 30 minutes.

[0417] 1.5 Data Analysis

[0418] The average NAD signal from the no-enzyme wells was subtracted from the fluorescence signal of each assay well to calculate the signal loss due to NAD+ depletion caused by PARP1 enzyme activity.

[0419] The % enzyme activity in each assay well (at varying compound concentrations) was calculated relative to the no compound / DMSO well (maximum enzyme activity well).

[0420] Graphpad Prism software was used to perform nonlinear regression curve fitting of "Sigmoidal dose response (variable slope)"; 4 parameters and slope to obtain IC50 values ​​of compounds.

[0421] constraint:

[0422] Bottom = constant equal to 0;

[0423] Top = must be less than 120.

[0424] 2. Determination of the in vitro inhibitory activity (IC50 value) of different compounds against ULK1

[0425] 2.1 Experimental Methods

[0426] The substrate solution was prepared by adding the substrate poly(Glu, Tyr) sodium salt (Sigma Aldrich, St. Louis, MO) to substrate reaction buffer (20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg / mL BSA, 0.1 mM Na3VO4, 2 mM DTT, and 1% DMSO) (the final substrate concentration in the reaction was 0.2 μM). Test compounds (compounds prepared in Examples 26 to 59, respectively) were prepared in 100% DMSO to a 10 mM stock solution and serially diluted 3-fold in 10 doses in a 384-well cyclic olefin copolymer LDV microplate. ULK1 kinase (recombinant human full-length protein, histidine-tagged, expressed in insect cells, Invitrogen, Carlsbad, CA) was added to the substrate solution and gently mixed (the final ULK1 concentration in the reaction was 8 nM). Next, 100% DMSO containing the test compound (compounds prepared in Examples 26 to 59, respectively) was added to the kinase reaction mixture using acoustic liquid transfer technology (Echo 550; nanoliter range) (Labcyte Inc., Sunnyvale, CA) and incubated at room temperature for 20 minutes. 33P-ATP (specific activity 10 μCi / μl) was added to the reaction mixture to initiate the reaction, followed by incubation at room temperature for 2 hours. A small aliquot of the reaction solution was spotted onto P-81 ion exchange filter paper (Whatman). Unbound phosphate was washed off the filter paper three times with 0.75% phosphate buffer and dried, and the radioactivity remaining on the filter paper was measured.

[0427] 2.2 Data Analysis

[0428] ULK1 kinase activity data were expressed as the percentage of the remaining kinase activity in the test sample relative to the vehicle (dimethyl sulfoxide) blank reaction. The data were curve fitted using Prism (GraphPad Software) to calculate the IC50 value.

[0429] 3. Experimental results

[0430] Table 1. Inhibition results of Parp1 and ULK1 enzyme activities Note: A: Enzyme activity inhibition ≤ 100nM; B: 100nM < Enzyme activity inhibition ≤ 1uM; C: 1uM < Enzyme activity inhibition.

[0431] As shown in Table 1, most of the representative compounds of the present invention (Compound I-1 to Compound I-34) showed strong inhibitory effects on Parp1 and ULK1 (<100 nM).

[0432] Example 61: Determination of In Vitro Huh-7 and MDA-MB-468 Cell Proliferation Inhibitory Activity (IC50 Value)

[0433] 1. Experimental methods

[0434] (i) Resuscitate Huh-7 and MDA-MB-468 cells and passage them twice or more. Harvest the cells to prepare a single-cell suspension and seed the cells into a transparent 96-well plate according to the cell pattern, with 10 cells per well.

[0435] (ii) The next day, drug working solutions were prepared using complete culture medium, and the test drugs (Compound I-11 prepared in Example 36, Compound I-9 prepared in Example 34, Compound I-14 prepared in Example 39, and commercially available positive control compound Olaparib) were added according to the cell layout. The highest drug concentration was 10 μM, and the cells were diluted 3.16-fold for a total of 9 concentrations, with 3 replicates for each concentration. A DMSO control was also set up.

[0436] (iii) placing the 96-well plate in a CO2 incubator and culturing at 37°C for 120 h;

[0437] (iv) After the culture was completed, an equal volume of cell culture medium was added to each well. Reagent;

[0438] (v) Mixing for 2 minutes using an orbital shaker to induce cell lysis;

[0439] (vi) Incubate the plate at room temperature for 10 minutes to allow the luminescence signal to stabilize;

[0440] (vii) The supernatant was transferred to a white opaque 96-well plate and the luminescence signal was recorded using a microplate reader.

[0441] 2. Experimental results

[0442] The test compounds (compound I-11 prepared in Example 36, compound I-9 prepared in Example 34, and compound I-14 prepared in Example 39, respectively) and the positive control compound Olaparib were incubated with MDA-MB-468 and Huh-7 cells, respectively, for 120 hours. The cell viability was detected by the CTG method. With drug concentration as the X-axis and cell viability as the Y-axis, the data were fitted using nonlinear S-curve regression to obtain a dose-effect curve, and the IC50 was calculated.

[0443] Table 2. IC50 values ​​of test compounds on cells

[0444] As shown in Table 2, some of the compounds described in the present invention showed stronger growth inhibitory effects on cancer cells MDA-MB-468 and Huh-7 than the reference compound Olaparib.

[0445] The present invention provides a PARP1 and ULK1 dual-target inhibitor, its preparation method, and its application. There are many specific methods and approaches for implementing this technical solution. The above is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and such improvements and modifications should also be considered within the scope of protection of the present invention. Any components not specified in this embodiment can be implemented using existing technologies.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof; Among them, R 1 Selected from halogen, cyano, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy-substituted C 1-6 alkyl, alkoxy-substituted C 1-6 alkyl, C 3-6 cycloalkyl, halo C 3-6 cycloalkyl or C 3-6 heterocycloalkyl; R 2 selected from -NR 5 R 6 、-NHNR 5 R 6 、-NR 5 OR 6 、-OR 5 、-ONR 5 R 6 or -SR 5 ; wherein, R 5 and R 6 are each independently selected from H, substituted or unsubstituted C 6-10 aryl, substituted or unsubstituted C 3-9 heteroaryl, substituted or unsubstituted C 1-3 alkyl, substituted or unsubstituted C 3-8 cycloalkyl, substituted or unsubstituted C 3-7 heterocycloalkyl, or R 5 and R 6 together with the N in -NR 5 R 6 form a 3- to 6-membered heterocycle; wherein the substitution is selected from being substituted by halogen, cyano, hydroxy, amino, mono(C 1-3 alkyl)amino, di(C 1-3 alkyl)amino, C 3-7 cycloamino, C 3-7 heterocycloamino, C 4-7 cycloamido, C 1-3 alkyl, halo C 1-3 alkyl, C 1-3 alkoxy or halo C 1-3 alkoxy substitution; R 3 Any one selected from the following groups: wherein, R 4 is selected from H, halogen, cyano, C 1-6 alkyl, halo-C 1-6 alkyl, C 1-3 alkoxy or halo-C 1-3 alkoxy; The L ring is selected from a substituted or unsubstituted 5- to 6-membered aromatic ring, or a substituted or unsubstituted 5- to 6-membered heteroaromatic ring; wherein, the substitution is selected from being substituted by halogen, cyano, hydroxyl, amino, mono(C 1-3 alkyl)amino, di(C 1-3 alkyl)amino, C 1-3 alkyl, halo C 1-3 alkyl, C 1-3 alkoxy or halo C 1-3 alkoxy substitution; W is selected from N or CH.

2. The compound of formula I as described in claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, characterized in that, R 1 Selected from halogen, C 1-3 alkyl, halo-C 1-3 alkyl, C 3-6 cycloalkyl or C 3-6 heterocycloalkyl.

3. The compound of formula I as described in claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, characterized in that, R 2 selected from -NR 5 R 6 or -OR 5 ; wherein, R 5 and R 6 are each independently selected from H, substituted or unsubstituted C6 aryl, substituted or unsubstituted C 3-5 heteroaryl, substituted or unsubstituted C 1-3 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted C 3-6 heterocycloalkyl, or R 5 and R 6 together with the N in -NR 5 R 6 form a 3- to 6-membered heterocycle; wherein, the said substitution is selected from being substituted by halogen, cyano, hydroxyl, amino, mono(C 1-3 alkyl)amino, di(C 1-3 alkyl)amino, C 4-7 cyclic amide group, C 1-3 alkyl, halo C 1-3 alkyl, C 1-3 alkoxy or halo C 1-3 alkoxy substitution.

4. The compound of formula I as described in claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, characterized in that, R 3 Selected from any one of the following groups: Among them, R 4 is selected from H or a halogen.

5. The compound of formula I as described in claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, characterized in that, The L ring is selected from a substituted or unsubstituted 5- or 6-membered aromatic ring, or a substituted or unsubstituted 5- or 6-membered heteroaromatic ring; wherein, the substitution is selected from being substituted by a halogen.

6. The compound of formula I as defined in claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, characterized in that, W is selected from N.

7. The compound of formula I as described in claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, characterized in that, R 1 Selected from halo C 1-3 alkyl; R 2 selected from -NR 5 R 6 ; wherein, R 5 and R 6 are each independently selected from H, substituted or unsubstituted C 1-3 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted C 3-6 heterocycloalkyl; wherein, the substitution is selected from being substituted by an amino group or a pyrrolidone group; R 3 Selected from any one of the following groups: wherein, R 4 is selected from H or fluorine; The L ring is selected from a substituted or unsubstituted 6-membered aromatic ring, or an unsubstituted 6-membered heteroaromatic ring; wherein, the substitution is selected from being substituted by fluorine; W is selected from N; Preferably, R 1 selected from trifluoromethyl; R 2 Selected from any one of the following groups: R 3 Any one selected from the following groups: The L ring is selected from any one of the following groups: W is selected from N.

8. The compound of formula I or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof according to claim 1, characterized in that, The compound described is selected from any one of the following structures:

9. A process for preparing the compound according to any one of claims 1 to 8, characterized in that, Comprising the following steps: Step 1: Nucleophilic substitution of compound A-1 with compound B to obtain compound A-2; Step 2: Compound A-2 is coupled with compound C to form compound I; wherein, the compound B is selected from amines, hydroxylamines, hydrazines, alcohols or thiols; the compound C is Among them, R 1 Selected from halogen, cyano, C 1-6 alkyl, halo-C 1-6 alkyl, hydroxy-substituted C 1-6 alkyl, alkoxy-substituted C 1-6 alkyl, C 3-6 cycloalkyl, halo-C 3-6 cycloalkyl or C 3-6 heterocycloalkyl; R 2 selected from -NR 5 R 6 、-NHNR 5 R 6 、-NR 5 OR 6 、-OR 5 、-ONR 5 R 6 or -SR 5 ; wherein, R 5 and R 6 are each independently selected from H, substituted or unsubstituted C 6-10 aryl, substituted or unsubstituted C 3-9 heteroaryl, substituted or unsubstituted C 1-3 alkyl, substituted or unsubstituted C 3-8 cycloalkyl, substituted or unsubstituted C 3-7 heterocycloalkyl, or R 5 and R 6 together with the N in -NR 5 R 6 form a 3- to 6-membered heterocycle; wherein, the said substitution is selected from being substituted by halogen, cyano, hydroxy, amino, mono(C 1-3 alkyl)amino, di(C 1-3 alkyl)amino, C 3-7 cyclic amino, C 3-7 heterocyclic amino, C 4-7 cyclic amide, C 1-3 alkyl, halo C 1-3 alkyl, C 1-3 alkoxy or halo C 1-3 alkoxy substitution; R 3 Any one selected from the following groups: wherein, R 4 is selected from H, halogen, cyano, C 1-6 alkyl, halo C 1-6 alkyl, C 1-3 alkoxy or halo C 1-3 alkoxy; The L ring is selected from a substituted or unsubstituted 5- to 6-membered aromatic ring or a substituted or unsubstituted 5- to 6-membered heteroaromatic ring; wherein the substitution is selected from being substituted by halogen, cyano, hydroxy, amino, mono(C 1-3 alkyl)amino, di(C 1-3 alkyl)amino, C 1-3 alkyl, halo C 1-3 alkyl, C 1-3 alkoxy or halo C 1-3 alkoxy substitution; W is selected from N or CH.

10. A pharmaceutical composition, characterized in that, Comprising the compound of formula I as described in any one of claims 1 to 8 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, and one or more pharmaceutically acceptable carriers.

11. A pharmaceutical preparation, characterized in that, It comprises a therapeutically effective amount of the compound of formula I as described in any one of claims 1 to 8 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, optical isomer or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

12. Use of the compound as described in any one of claims 1 to 8 or the pharmaceutical composition as described in claim 10 or the pharmaceutical preparation as described in claim 11 in the preparation of a poly(ADP-ribose) polymerase 1 inhibitor.

13. Use of the compound as described in any one of claims 1 to 8 or the pharmaceutical composition as described in claim 10 or the pharmaceutical preparation as described in claim 11 in the preparation of an unc-51-like autophagy activating kinase 1 inhibitor.

14. Use of the compound as described in any one of claims 1 to 8 or the pharmaceutical composition as described in claim 10 or the pharmaceutical preparation as described in claim 11 in the preparation of a drug for preventing or treating a disease related to abnormal enzyme activity of poly(ADP-ribose) polymerase 1 and / or unc-51-like autophagy activating kinase 1.

15. The use according to claim 14, wherein, The disease related to abnormal enzyme activity of poly(ADP-ribose) polymerase 1 and / or unc-51-like autophagy activating kinase 1 is a tumor.

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