Pharmaceutical Use of a Class of Substituted Heteroarylphthalazine Derivatives and Preparation Method Thereof

Abstract

The present disclosure relates to a substituted heteroaryl phthalazine derivative represented by formula (I-0), its use as an NLRP3 inhibitor and a preparation method thereof, and it has good NLRP3 inhibitory activity.

Description

Technical Field

[0001] The present disclosure relates to the field of medicine, and particularly to NLRP3 inhibitors comprising a compound represented by chemical formula (I) or a pharmaceutically acceptable salt thereof, their uses and preparation methods. Background Art

[0002] Inflammation is a defensive response of the body to stimuli, including infectious inflammation and sterile inflammation. Inflammation is mainly manifested as redness, swelling, heat, pain and dysfunction, which is caused by the increased permeability of vascular endothelial cells and the exudation of immune cells in plasma. After tissue repair, the inflammatory response will end quickly, but excessive cytokine production will lead to a cytokine storm, causing damage to the body. Inflammatory dysregulation is an important pathogenesis of many human diseases.

[0003] Innate immunity plays a crucial role in inflammation. Cells such as macrophages, dendritic cells, epithelial cells, endothelial cells, and fibroblasts are all involved in the innate immune response. Innate immunity recognizes pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPs) through pattern recognition receptors. Currently, five classes of pattern recognition receptors have been identified, including Toll-like receptors (TLRs), C-type lectin receptors (CLRs), RIG-like receptors (RLRs), cytoplasmic DNA sensors, and NOD-like receptors (NLRs). These pattern recognition receptors are expressed on both immune cells and non-immune cells. After the pattern recognition receptors recognize the corresponding ligands, they initiate and activate multiple innate immune signaling pathways, generating a series of cytokines that promote inflammation and type I interferons.

[0004] NLRs are a class of intracellular pattern recognition receptors. NLRs are widely expressed in various immune cells and epithelial cells and can initiate innate immune responses by recognizing PAMPs entering the cell and DAMPs such as cellular stress. The high evolutionary conservation of NLRs also proves their crucial role in maintaining immune homeostasis in the body. NLRs regulate inflammatory responses by promoting the production of cytokines, chemokines, and the expression of antibacterial-related genes. Therefore, NLRs are closely related to human diseases such as infections, tumors, autoimmune diseases, and inflammatory disorders.

[0005] NLRs are composed of an LRR domain at the C-terminus, a NOD domain in the middle, and an effector domain at the N-terminus. The LRR domain at the C-terminus is responsible for recognizing and binding ligands. The NOD domain in the middle has dNTPase (deoxynucleosidetriphosphohydrolase) enzyme activity and is responsible for the oligomerization of NLRs proteins. The effector domain at the N-terminus exerts effector functions by interacting with other proteins. Four types of N-terminal effector domains have been discovered so far, namely the AD domain (acidic transactivation domain), the BIR domain (baculoviral inhibitory repeat-like domain), the CARD domain (caspase activation and recruitment domain), and the PYD domain (pyrin domain). According to the different N-terminal effector domains, NOD-like receptors can be divided into four subfamilies, namely the NLRA subfamily (containing the Acidic domain), the NLRB subfamily (containing the BIR domain), the NLRC subfamily (containing the CARD domain), and the NLRP subfamily (containing the pyrin domain). The NLRP subfamily includes 14 members, namely NLRP1-14.

[0006] The inflammasome is a class of multi-protein complexes that can mediate the activation of caspase-1. Activated caspase-1 promotes the processing and maturation of inflammatory cytokines IL-1β and IL-18 and also leads to the occurrence of pyroptosis. Pyroptosis results in the release of more DAMPs, further enhancing the immune response. Eight NLRs have been found to form inflammasomes after activation, namely NLRP1, NLRP2, NLRP3, NLRP6, NLRP7, NLRP12, NLRC4, and NAIP.

[0007] The NLRP3 inflammasome is composed of the pattern recognition receptor NLRP3, the adaptor protein ASC, and the effector protein pro-caspase-1. NLRP3 consists of an N-terminal PYD domain, a middle NACHT domain, and a C-terminal LRR domain. Activation of the NLRP3 inflammasome leads to the production of active caspase-1, which further promotes pyroptosis. The NLRP3 inflammasome can recognize a variety of stimulants, including protozoa (Plasmodium, amoeba, etc.), viruses (adenovirus, influenza virus, Sendai virus, etc.), fungi (Saccharomyces cerevisiae, Candida albicans, etc.), bacteria (Listeria, Escherichia coli, Staphylococcus aureus, etc.). NLRP3 can also recognize many endogenous DAMPs, including uric acid crystals, ATP, islet amyloid polypeptide, β-amyloid plaques, etc.

[0008] In addition, the abnormal activation of the NLRP3 inflammasome is an important promoting factor in many major human diseases such as rheumatoid arthritis, gouty arthritis, atherosclerosis, myocardial infarction, Parkinson's syndrome, Alzheimer's disease, infectious lung injury, pulmonary fibrosis, sepsis, ulcerative colitis, type 2 diabetes, sepsis, bacterial inflammation, familial Mediterranean fever, nephrotic syndrome, myocarditis, etc. Therefore, NLRP3 inhibitors have certain therapeutic potential in these diseases with inflammatory pathological characteristics. Summary of the Invention

[0009] The purpose of the present disclosure is to provide a substituted heteroaryl phthalazine derivative with NLRP3 inhibitory activity.

[0010] The present disclosure provides a compound represented by the following chemical formula (I-0) or a pharmaceutically acceptable salt thereof:

[0011]

[0012] Wherein:

[0013] n is 0 or 1;

[0014] m is selected from an integer of 1 to 5;

[0015] p is selected from 1 or 2;

[0016] X1, X2, and X5 are each independently selected from CH2, NH, CH, O, S, or N;

[0017] X3 and X4 are each independently selected from CH2, CH, or N;

[0018] R1 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, halogen, phosphinyl, hydroxy, cyano, the C 1-6alkyl, C 3-6 cycloalkyl, C 1-6 alkoxy, phosphinyl optionally substituted by one or more halogens, C 1-3 alkyl, wherein m R1s may be the same as or different from each other;

[0019] R3 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, halogen, phosphinyl, carboxyl, cyano, said C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, phosphinyl optionally substituted by one or more halogens, C 1-3 alkyl, wherein p R3s may be the same as or different from each other;

[0020] A is a single bond or a C 1-3 alkylene chain, optionally, one or more hydrogens on the methylene in said C 1-3 alkylene chain are substituted by C 1-3 alkyl;

[0021] M is -NR 10 -, -O- or -S-

[0022] R4 is selected from C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3-9 membered heterocycloalkyl, 5-9 membered heteroaryl, 9-12 membered partially unsaturated heterocyclic bicyclic, said C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3-9 membered heterocycloalkyl, 5-9 membered heteroaryl, 9-12 membered partially unsaturated heterocyclic bicyclic optionally substituted by one or more halogens, hydroxyl, C 1-3 alkyl, C 1-6 acyl, =O, -NR8R9;

[0023] R8, R9, R 10 are each independently selected from hydrogen, C 1-3 alkyl.

[0024] Specifically, the present disclosure relates to a compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof,

[0025]

[0026] wherein:

[0027] n is 0 or 1;

[0028] m is an integer selected from 1 to 5;

[0029] p is selected from 1 or 2;

[0030] X1, X2, and X5 are each independently selected from CH2, NH, CH, O, S, or N;

[0031] X3 and X4 are each independently selected from CH2, CH, or N;

[0032] R1 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, halogen, phosphinyl, hydroxy, cyano, wherein the C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkoxy, phosphinyl is optionally substituted with one or more halogens, C 1-3 alkyl, where m R1s may be the same or different from each other;

[0033] R3 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, halogen, phosphinyl, carboxy, cyano, wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, phosphinyl is optionally substituted with one or more halogens, C 1-3 alkyl, where p R3s may be the same or different from each other;

[0034] A is a single bond or a C 1-3 alkylene chain, optionally, one or more hydrogens on the methylene groups in the C 1-3 alkylene chain are substituted with C 1-3 alkyl;

[0035] R4 is selected from C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3-9 membered heterocycloalkyl, 5-9 membered heteroaryl, 9-12 membered partially unsaturated heterocyclic bicyclic, wherein the C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3-9 membered heterocycloalkyl, 5-9 membered heteroaryl, 9-12 membered partially unsaturated heterocyclic bicyclic is optionally substituted with one or more halogens, hydroxy, C 1-3 alkyl, C 1-6 acyl, =O, -NR8R9;

[0036] R8 and R9 are each independently selected from hydrogen, C 1-3 alkyl.

[0037] Preferably, R4 is selected from C1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3- to 9-membered heterocycloalkyl, 5- to 9-membered heteroaryl, 9- to 12-membered partially unsaturated heterocyclic bicyclic, said C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3- to 9-membered heterocycloalkyl, 5- to 9-membered heteroaryl, 9- to 12-membered partially unsaturated heterocyclic bicyclic optionally substituted by one or more halogens, hydroxyl, C 1-3 alkyl, C 1-6 acyl, -NR8R9.

[0038] Preferably, R1 is selected from C 1-3 alkyl, C 1-3 alkoxy, C 3-6 cycloalkyl, phosphinyl, hydroxyl, cyano, halogen, said C 1-3 alkyl, C 1-3 alkoxy, C 3-6 cycloalkyl, phosphinyl optionally substituted by one to three halogens, C 1-3 alkyl.

[0039] Preferably, R1 is selected from C 1-3 alkyl, C 1-3 alkoxy, hydroxyl, cyano, halogen, phosphinyl, said C 1-3 alkyl, C 1-3 alkoxy, phosphinyl optionally substituted by one to three fluorines, methyl.

[0040] Preferably, R1 is selected from trifluoromethyl, difluoromethyl, methyl, fluorine, hydroxyl, dimethylphosphinyl or trifluoromethoxy.

[0041] Preferably, R1 is selected from trifluoromethyl, methyl, fluorine, hydroxyl, dimethylphosphinyl or trifluoromethoxy.

[0042] Preferably, R1 is selected from trifluoromethyl, methyl or hydroxyl.

[0043] Preferably, R3 is selected from hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, C 1-3 alkoxy, halogen, phosphinyl, said C 1-3 alkyl, C 1-3 alkoxy, C 3-6 cycloalkyl, phosphinyl optionally substituted by one to three halogens, C 1-3 alkyl.

[0044] Preferably, R3 is selected from C 1-3 alkyl, C 1-3 alkoxy, C 3-6Cycloalkyl, phosphine oxide group, said C 1-3 alkyl, C 1-3 alkoxy, C 3-6 cycloalkyl, phosphine oxide group is optionally substituted by one to three fluorine, methyl.

[0045] Preferably, R3 is selected from hydrogen, methyl, methoxy, cyclopropyl, ethyl, fluorine, trifluoromethyl or dimethylphosphine oxide group.

[0046] Preferably, R3 is selected from hydrogen, methyl, methoxy.

[0047] Preferably, A is a single bond or C 1-3 alkylene chain, optionally, one or more hydrogens on the methylene in the C 1-3 alkylene chain are substituted by methyl.

[0048] Preferably, A is a single bond, -CH2-, -(CH3)CH-, -CH2CH2-.

[0049] Preferably, A is a single bond.

[0050] Preferably, R4 is selected from C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5-7 membered heterocycloalkyl containing 1-2 atoms independently selected from N, O, S atoms respectively, 5-7 membered heteroaryl containing 1-2 atoms independently selected from N, O, S atoms respectively, 9-12 membered partially unsaturated heterocyclic bicyclic containing 1-2 atoms independently selected from N, O, S atoms respectively, said C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5-7 membered heterocycloalkyl containing 1-2 atoms independently selected from N, O, S atoms respectively, 5-7 membered heteroaryl containing 1-2 atoms independently selected from N, O, S atoms respectively, 9-12 membered partially unsaturated heterocyclic bicyclic containing 1-2 atoms independently selected from N, O, S atoms respectively is optionally substituted by one or more halogens, hydroxyl, C 1-3 alkyl, C 1-6 acyl, halo C 1-3 alkyl, =O, -NR8R9.

[0051] Preferably, R4 is selected from C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5-7 membered heterocycloalkyl containing 1-2 atoms independently selected from N, O, S atoms respectively, 5-7 membered heteroaryl containing 1-2 atoms independently selected from N, O, S atoms respectively, 9-12 membered partially unsaturated heterocyclic bicyclic containing 1-2 atoms independently selected from N, O, S atoms respectively, said C 1-6 alkyl, C 5-8Cycloalkyl, phenyl, 5- to 7-membered heterocycloalkyl containing 1 to 2 atoms independently selected from N, O, S atoms, 5- to 7-membered heteroaryl containing 1 to 2 atoms independently selected from N, O, S atoms, 9- to 12-membered partially unsaturated heterocyclic bicyclic containing 1 to 2 atoms independently selected from N, O, S atoms, optionally substituted by one or more halogens, hydroxyl groups, C 1-3 alkyl, C 1-6 acyl, -NR8R9.

[0052] Preferably, R4 is selected from C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5- to 7-membered heterocycloalkyl containing 1 to 2 atoms independently selected from N, O atoms, 5- to 7-membered heteroaryl containing 1 N atom, 9- to 12-membered partially unsaturated heterocyclic bicyclic containing 1 N atom, the C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5- to 7-membered heterocycloalkyl containing 1 to 2 atoms independently selected from N, O atoms, 5- to 7-membered heteroaryl containing 1 N atom, 9- to 12-membered partially unsaturated heterocyclic bicyclic containing 1 N atom, optionally substituted by one or more halogens, hydroxyl groups, C 1-3 alkyl, C 1-6 acyl, halo C 1-3 alkyl, =O, -NR8R9.

[0053] Preferably, R4 is selected from C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5- to 7-membered heterocycloalkyl containing 1 to 2 atoms independently selected from N, O atoms, 5- to 7-membered heteroaryl containing 1 N atom, 9- to 12-membered partially unsaturated heterocyclic bicyclic containing 1 N atom, the C 1-6 alkyl, C 5-8 cycloalkyl, phenyl, 5- to 7-membered heterocycloalkyl containing 1 to 2 atoms independently selected from N, O atoms, 5- to 7-membered heteroaryl containing 1 N atom, 9- to 12-membered partially unsaturated heterocyclic bicyclic containing 1 N atom, optionally substituted by one or more halogens, hydroxyl groups, C 1-3 alkyl, C 1-6 acyl, -NR8R9.

[0054] Preferably, R4 is selected from n-butyl, cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolyl, pyrrolidinyl, morpholinyl, tetrahydropyranyl, the cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolyl, pyrrolidinyl optionally substituted by one or more halogens, hydroxyl groups, C 1-3 alkyl, C 1-6 acyl, halo C 1-3 alkyl, =O, -NR8R9.

[0055] Preferably, R4 is selected from n-butyl, cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolyl, pyrrolidinyl, morpholinyl, the cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolyl, pyrrolidinyl are optionally substituted by one or more halogens, hydroxyl groups, C 1-3 alkyl, C 1-6 acyl, -NR8R9.

[0056] Preferably, R4 is selected from n-butyl, cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolidinyl, morpholinyl, tetrahydropyranyl, the cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolidinyl are optionally substituted by one to two fluorines, hydroxyl groups, methyl, ethyl, acetyl, halo C 1-3 alkyl, =O, -N(CH3)2.

[0057] Preferably, R4 is selected from n-butyl, cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolidinyl, morpholinyl, the cyclohexyl, phenyl, piperidinyl, pyridinyl, pyrrolidinyl are optionally substituted by one to two fluorines, hydroxyl groups, methyl, ethyl, acetyl, -N(CH3)2.

[0058] Preferably, R4 is selected from n-butyl, -C(CH3)2OH, phenyl,

[0059] Preferably, R4 is selected from n-butyl, -C(CH3)2OH, phenyl,

[0060] Preferably, R4 is selected from

[0061] Preferably, R8 and R9 are methyl.

[0062] The present disclosure relates to a compound having the structure shown in formula (II) or a pharmaceutically acceptable salt thereof:

[0063]

[0064] Wherein:

[0065] The definitions of n, X1, X2, X3, X4, X5, R3, R4, and A are as defined above.

[0066] Preferably, R 11 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, halogen, phosphinyl oxide, and the C 1-6 alkyl, C 3-6Cycloalkyl, C 1-6 Alkoxy, phosphine oxide group is optionally substituted by one or more halogens, C 1-3 Alkyl.

[0067] Preferably, R 11 Is selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, phosphine oxide group, halogen, C 3-6 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy, C 3-6 Cycloalkyl, phosphine oxide group is optionally substituted by one to three halogens, C 1-3 Alkyl.

[0068] Preferably, R 11 Is selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, halogen, phosphine oxide group, the C 1-3 Alkyl, C 1-3 Alkoxy, phosphine oxide group is optionally substituted by one to three fluorines, methyl.

[0069] Preferably, R 11 Is selected from hydrogen, trifluoromethyl, methyl, fluorine, dimethylphosphine oxide group or trifluoromethoxy.

[0070] Preferably, R 11 Is selected from trifluoromethyl, methyl, trifluoromethoxy, fluorine.

[0071] R2 is selected from hydrogen, deuterium, C 1-6 Alkyl, hydroxy, halogen, cyano, difluoromethyl.

[0072] Preferably, R2 is selected from hydrogen, deuterium, C 1-6 Alkyl, hydroxy, halogen, cyano.

[0073] Preferably, R2 is selected from hydroxy, difluoromethyl.

[0074] Preferably, R2 is selected from hydroxy.

[0075] R5, R6, R7 are each independently selected from hydrogen, halogen, C 1-3 Alkyl.

[0076] Preferably, R5, R6, R7 are each independently selected from hydrogen, fluorine, methyl.

[0077] This disclosure relates to a compound having the structure shown in formula (III) or a pharmaceutically acceptable salt thereof:

[0078]

[0079] Wherein:

[0080] n, X1, X2, R 11 , R3, R4, A are as defined above.

[0081] The present disclosure relates to a compound having the structure shown in formula (IV) or a pharmaceutically acceptable salt thereof:

[0082]

[0083] Wherein:

[0084] X1, X2, R 11 , R3, R4, A are as defined above.

[0085] The present disclosure also relates to a compound having the structure shown in formula (IVa), (IVb), (IVc), (IVd) or (IVe) or a pharmaceutically acceptable salt thereof:

[0086]

[0087] Wherein:

[0088] R1, R3, R4, A are as defined above.

[0089] The present disclosure further relates to a compound having the structure shown in formula (V) or formula (VI) or a pharmaceutically acceptable salt thereof:

[0090]

[0091] Wherein:

[0092] R3, R4, A are as defined above. The present disclosure relates to a compound having the structure shown in formula (I-1) or a pharmaceutically acceptable salt thereof:

[0093]

[0094] Wherein:

[0095] n is 0 or 1;

[0096] m is selected from the integers from 1 to 5;

[0097] p is selected from 1 or 2;

[0098] X1, X2, X5, X6 are each independently selected from CH2, NH, CH, O, S or N;

[0099] X3, X4 are each independently selected from CH2, CH or N;

[0100] R1 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6Cycloalkyl, halogen, phosphinyl, hydroxy, cyano, said C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkoxy, phosphinyl optionally substituted by one or more halogens, C 1-3 alkyl, wherein m R1s may be the same as or different from each other;

[0101] R3 is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, halogen, phosphinyl, carboxy, cyano, said C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, phosphinyl optionally substituted by one or more halogens, C 1-3 alkyl, wherein p R3s may be the same as or different from each other;

[0102] A is a single bond or a C 1-3 alkylene chain, optionally, one or more hydrogens on the methylene in the C 1-3 alkylene chain are substituted by C 1-3 alkyl;

[0103] R4 is selected from C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3-9-membered heterocycloalkyl, 5-9-membered heteroaryl, 9-12-membered partially unsaturated heterocyclic bicyclic, said C 1-6 alkyl, C 3-9 cycloalkyl, C 5-9 aryl, 3-9-membered heterocycloalkyl, 5-9-membered heteroaryl, 9-12-membered partially unsaturated heterocyclic bicyclic optionally substituted by one or more halogens, hydroxy, C 1-3 alkyl, C 1-6 acyl, =O, -NR8R9;

[0104] R8 and R9 are each independently selected from hydrogen, C 1-3 alkyl;

[0105] Preferably, X6 is selected from S.

[0106] This disclosure relates to the following compounds or pharmaceutically acceptable salts thereof:

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115] The present disclosure relates to a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof, and the method comprises the following steps:

[0116]

[0117] Step 1: Dissolve compound A0 in POCl3, heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution. After removing POCl3, slowly drop the oily crude product into ice water, extract with ethyl acetate, and separate by column chromatography to obtain the target compound A1;

[0118] Step 2: Dissolve compound A1, the corresponding amine, and Na2CO3 in dry DMF. Place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water, extract with ethyl acetate, and separate by column chromatography to obtain the target compound A2;

[0119] Step 3: Add compound A2, boric acid, sodium carbonate, and Pd(dppf)Cl2 to a mixed solvent of dioxane and water. Replace with nitrogen 3 times, heat to 110 °C and react for 3 hours. Add the reaction solution to water, extract with ethyl acetate, and separate by column chromatography to obtain the target compound I-0.

[0120] In one embodiment, the method comprises the following steps:

[0121]

[0122] Step 1: Dissolve compound A0' in POCl3, heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution. After removing POCl3, slowly drop the oily crude product into ice water, extract with ethyl acetate, and separate by column chromatography to obtain the target compound A1';

[0123] Step 2: Dissolve compound A1', the corresponding amine, and Na2CO3 in dry DMF. Place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water, extract with ethyl acetate, and separate by column chromatography to obtain the target compound A2';

[0124] Step 3: Add compound A2’, boric acid, sodium carbonate and Pd(dppf)Cl2 into a mixed solvent of dioxane and water, displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Add the reaction solution into water, extract with ethyl acetate, and obtain the target compound II through column chromatography separation.

[0125] On the other hand, the present disclosure provides stereoisomers, solvates or prodrugs of any of the foregoing compounds.

[0126] The present disclosure relates to a pharmaceutical composition, which comprises any of the foregoing compounds or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0127] The present disclosure relates to the use of any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the foregoing pharmaceutical composition in the preparation of a drug for treating NLRP3-mediated diseases.

[0128] The present disclosure relates to the use of any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the foregoing pharmaceutical composition in the preparation of an inhibitor of NLRP3.

[0129] The present disclosure relates to a method for inhibiting NLRP3 in a patient in need thereof, which comprises administering to the patient any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the foregoing pharmaceutical composition.

[0130] The present disclosure relates to a method for treating NLRP3-mediated diseases in a patient in need thereof, which comprises administering to the patient any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the foregoing pharmaceutical composition. Preferably, the NLRP3-mediated diseases include but are not limited to rheumatoid arthritis, gouty arthritis, atherosclerosis, myocardial infarction, Parkinson's syndrome, Alzheimer's disease, infectious lung injury, pulmonary fibrosis, sepsis, ulcerative colitis, type 2 diabetes, sepsis, bacterial inflammation, familial Mediterranean fever, nephrotic syndrome, myocarditis.

[0131] On the other hand, the present disclosure provides the use of an NLRP3 inhibitor comprising any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition in the treatment of heart diseases.

[0132] On the other hand, the present disclosure provides the use of an NLRP3 inhibitor comprising any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition in the treatment of inflammatory diseases.

[0133] On the other hand, the present disclosure provides the use of an NLRP3 inhibitor comprising any of the foregoing compounds or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition in the treatment of infectious diseases.

[0134] The compounds of the present disclosure have NLRP3 inhibitory effects. Detailed Description of the Invention

[0135] I. Definitions

[0136] According to the above content of the present disclosure, and based on common general knowledge and conventional means in the art, without departing from the above basic technical concept of the present disclosure, various other forms of modification, substitution or variation can be made.

[0137] Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "having" etc. will be understood to include the stated element or component, without excluding other elements or other components.

[0138] The compounds of the present disclosure may be asymmetric, for example, having one or more stereoisomers. Unless otherwise specified, all stereoisomers are included, such as enantiomers and diastereoisomers. Compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure form or in racemic form. The optically pure form can be resolved from the racemic mixture, or synthesized by using chiral starting materials or chiral reagents. Racemates, diastereoisomers and enantiomers are all included within the scope of the present disclosure.

[0139] In the present disclosure, refers to the position where the substituent is bonded

[0140] In the present disclosure, a numerical range refers to each integer within the given range. For example, "C1-6" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms; "C1-3" means that the group may have 1 carbon atom, 2 carbon atoms or 3 carbon atoms.

[0141] The term "optional" or "optionally" means that the subsequent described event or situation may or may not occur, and the description includes the occurrence and non-occurrence of the described event or situation.

[0142] The term "substituted" or "substitution" means that any one or more hydrogen atoms on a specific atom or group are replaced by substituents, provided that the valence of the specific atom or group is normal and the resulting compound is stable. When the substituent is a keto group (i.e., =O), it means that two hydrogen atoms are replaced. Unless otherwise specified, the type and number of substituents can be arbitrary on the basis of being chemically achievable. The substituents can be selected from one, two or more of the following substituents: deuterium, halogen group, cyano group, nitro group, -C(=O)R, -C(=O)OR', -OC(=O)R", imide group, amide group, hydroxyl group, substituted or unsubstituted amino group, substituted or unsubstituted alkyl group, substituted or unsubstituted cycloalkyl group, substituted or unsubstituted haloalkyl group, substituted or unsubstituted alkoxy group, substituted or unsubstituted alkenyl group, substituted or unsubstituted alkynyl group, substituted or unsubstituted aryl group, substituted or unsubstituted aryloxy group, substituted or unsubstituted heteroaryl group, etc., but not limited thereto.

[0143] When any variable (e.g., R n ) appears more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted by one to three Rs, the group can optionally be substituted by up to three Rs, and each R in each case has an independent option. In addition, combinations of substituents and / or their variants are permitted only if such combinations result in a stable compound.

[0144] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain or branched saturated hydrocarbon groups having the number of carbon atoms shown. As the term "C 1-6 alkyl" includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, and examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, 3-hexyl, etc. It can be divalent, such as methylene, ethylene.

[0145] The term "alkoxy" can be straight-chain, branched or cyclic. The number of carbon atoms in the alkoxy is not particularly limited, but is preferably 1 to 20. Specific examples thereof include methoxy, ethoxy, n-propoxy, isopropoxy, i-propyloxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, n-pentyloxy, neopentyloxy, isopentyloxy, n-hexyloxy, 3,3-dimethylbutoxy, 2-ethylbutoxy, n-octyloxy, n-nonyloxy, n-decyloxy, etc., but not limited thereto.

[0146] The term "phosphoryl group" is -P(=O)(R m)(R n ) structure, where R m and R n are the same as or different from each other, and each independently is hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. Specific examples of phosphine oxide groups include alkylphosphine oxide groups, arylphosphine oxide groups, etc. More specifically, they include diphenylphosphine oxide groups, dinaphthylphosphine oxide groups, etc., but are not limited thereto.

[0147] The term "alkylene" or "alkylene chain" refers to a fully saturated straight-chain or branched-chain divalent hydrocarbon chain group having one to twelve carbon atoms, and can generally be represented as C1-C12 alkylene. Preferred alkylene is C1-C6 alkylene, and more preferably C1-C4 alkylene. Non-limiting examples of C1-C12 alkylene include methylene, ethylene, propylene, n-butylene, vinyl, propenyl, n-butenyl, propynyl, n-butynyl, etc. The alkylene chain is connected to the rest of the molecule by a single bond and is connected to the said group by a single bond. The points where the alkylene chain is connected to the rest of the molecule and to the said group can be through one carbon or any two carbons within the chain. Unless otherwise specifically specified in this specification, the alkylene chain can be optionally substituted.

[0148] In the present disclosure, examples of halogen groups can include fluorine, chlorine, bromine or iodine.

[0149] In the present disclosure, the term "cycloalkyl" refers to a monocyclic saturated hydrocarbon system without heteroatoms and double bonds. For example, examples of the term "C 3-9 cycloalkyl" include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl.

[0150] In the present disclosure, the term "aryl" refers to a fully carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π-electron system, which is obtained by removing a hydrogen atom from a single carbon atom of the parent aromatic ring system. It includes bicyclic groups containing an aromatic ring fused to a saturated, partially unsaturated ring, or an aromatic carbocyclic ring. Specific examples thereof include phenyl or naphthyl, but are not limited thereto.

[0151] In the present disclosure, the term "heterocycloalkyl" refers to a 5-12 membered saturated non-aromatic system having ring carbon atoms and 1 to 2 ring heteroatoms. Specific examples of heterocyclic groups include piperidinyl or tetrahydropyrrolyl, but are not limited thereto.

[0152] In the present disclosure, the term "heteroaryl" refers to a monovalent aryl group containing at least one heteroatom independently selected from nitrogen, oxygen, and sulfur. The heteroaryl group can be monocyclic or a polycyclic system, such as bicyclic, where two or more rings exist in a fused, bridged, or spiro form, and at least one ring contains one or more heteroatoms. Specific examples of heteroaryl groups include pyridyl, thienyl, imidazolyl, pyrimidinyl, pyridyl, furyl, pyrazinyl, thiazolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, imidazopyridinyl, benzofuryl, pyridazinyl, isoindolyl, but are not limited thereto.

[0153] In the present disclosure, when the heteroaryl group is substituted, it may include but is not limited thereto.

[0154] The term "heterocycle" refers to a 5- to 12-membered saturated non-aromatic system having ring carbon atoms and 1 to 2 ring heteroatoms, where the heteroatoms are independently selected from nitrogen, sulfur, or oxygen atoms. In a heterocyclic group containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, provided that the valence allows. The heterocycle can be a monocyclic or polycyclic system, such as bicyclic, where two or more rings exist in a fused, bridged, or spiro form, and at least one ring contains one or more heteroatoms.

[0155] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double bond or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

[0156] Drug or pharmaceutical composition

[0157] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are suitable, within the scope of reasonable medical judgment, for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

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

[0159] The pharmaceutically acceptable salts of the present disclosure can be synthesized from the parent compounds containing acid or base moieties by conventional chemical methods. Generally, the preparation method of such salts is to react these compounds in the form of free acid or base with a stoichiometric amount of an appropriate base or acid in water, an organic solvent, or a mixture of both.

[0160] The medicament or pharmaceutical composition of the present disclosure can be administered orally, topically, parenterally or mucosally (e.g., sublingually, by inhalation or rectally) in dosage unit formulations comprising conventional non-toxic pharmaceutically acceptable carriers.

[0161] For oral administration in the form of tablets or capsules, the active pharmaceutical ingredient can be combined with non-toxic, pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose); fillers (e.g., lactose, sucrose, glucose, mannitol, sorbitol and other reducing and non-reducing sugars, microcrystalline cellulose, calcium sulfate or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica, stearic acid, sodium stearyl fumarate, behenin, calcium stearate, etc.); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate), coloring agents and flavoring agents, gelatin, sweeteners, natural and synthetic gums (such as gum arabic, tragacanth or alginate), buffer salts, carboxymethylcellulose, polyethylene glycol, waxes, etc. For oral administration in liquid form, the pharmaceutical ingredient can be combined with non-toxic, pharmaceutically acceptable inert carriers (e.g., ethanol, glycerol, water), anti-settling agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats), emulsifiers (e.g., lecithin or gum arabic), non-aqueous carriers (e.g., almond oil, esters of fatty acids, ethanol or fractionated vegetable oils), preservatives (e.g., methyl p-hydroxybenzoate or propyl p-hydroxybenzoate or sorbic acid), etc. Stabilizers such as antioxidants (BHA, BHT, propyl gallate, sodium ascorbate, citric acid) can also be added to stabilize the dosage form.

[0162] Tablets containing the active compound can be coated by methods well known in the art. The compositions of the present disclosure containing the compound of formula I as the active compound can also be incorporated into beads, microspheres or microcapsules, for example constructed from polyglycolic acid / lactic acid (PGLA). Preparations for oral administration in liquid form can take, for example, the form of solutions, syrups, emulsions or suspensions or they can be presented as dry products to be reconstituted with water or other suitable excipients before use. Preparations for oral administration can be suitably formulated to provide controlled or delayed release of the active compound.

[0163] The medicaments or pharmaceutical compositions of the present disclosure can be delivered parenterally, i.e., by intravenous (i.v.), intracerebroventricular (i.c.v.), subcutaneous (s.c.), intraperitoneal (i.p.), intramuscular (i.m.), subdermal (s.d.) or intradermal (i.d.) administration, by direct injection, e.g., by bolus injection or continuous infusion. Formulations for injection can be presented in unit dosage forms, e.g., in ampoules or multi-dose containers with added preservatives. The compositions can be in the form of suspensions, solutions or emulsions in oily or aqueous carriers and can contain formulating agents such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient can be in powder form and reconstituted before use with a suitable carrier (e.g., sterile pyrogen-free water).

[0164] The medicaments or pharmaceutical compositions of the present disclosure can also be formulated for rectal administration, e.g., in the form of suppositories or retention enemas (e.g., containing conventional suppository bases such as cocoa butter or other glycerides).

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

[0166] The use of the terms "reduce", "inhibit", "alleviate" or "decrease" is relative to a control. A person skilled in the art will readily determine the appropriate control for each experiment. For example, the reduced response in a subject or cell treated with a compound is compared with the response in a subject or cell not treated with the compound.

[0167] As used herein, the term "effective amount" or "therapeutically effective amount" refers to a dose sufficient to treat, inhibit or alleviate one or more symptoms of the disease state being treated or otherwise provide the desired pharmacological and / or physiological effect. The exact dose will vary depending on a variety of factors such as subject-dependent variables (e.g., age, immune system health, etc.), the disease or disorder, and the treatment being administered. The effect of the effective amount can be relative to a control. Such controls are known in the art and are discussed herein and can be, for example, the condition of the subject before or without administration of the medicament or pharmaceutical combination, or in the case of a pharmaceutical combination, the combined effect can be compared with the effect of administering only one of the drugs.

[0168] The term "excipient" is used herein to include any other compound that is not a therapeutic or bioactive compound and can be included in or on the microparticles. Thus, the excipient should be pharmaceutically or biologically acceptable or relevant, e.g., the excipient is generally non-toxic to the subject. "Excipient" includes a single such compound and is also intended to include a plurality of compounds.

[0169] The term "pharmaceutical composition" means a composition comprising a compound described in the present disclosure or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable ingredient selected from the following, depending on the mode of administration and the nature of the dosage form, including but not limited to: carriers, diluents, adjuvants, excipients, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, dispersants, temperature-sensitive materials, temperature regulators, adhesives, stabilizers, suspending aids, etc.

[0170] Uses and methods of treatment

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

[0172] According to the methods of the present disclosure, the compounds or compositions can be administered in any amount and by any route of administration effective for treating a disease associated with NRLP3 or reducing its severity.

[0173] The present disclosure relates to a method of inhibiting NRLP3 in a biological sample, which comprises the step of contacting the biological sample with a compound of the present disclosure or a composition comprising the compound.

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

[0175] The method of inhibiting NRLP3 in a patient according to the present disclosure, which comprises the step of administering to the patient a compound of the present disclosure or a composition comprising the compound.

[0176] The compounds provided are NRLP3 inhibitors and can therefore be used to treat one or more conditions associated with NRLP3 activity. Thus, in certain embodiments, the present disclosure provides a method for treating NRLP3-mediated conditions, which comprises the step of administering to a patient in need thereof a compound of the present disclosure or a pharmaceutically acceptable composition thereof.

[0177] As used herein, the terms "NRLP3-mediated" conditions, diseases and / or disorders as used herein mean any disease or other adverse condition in which NRLP3 or a mutant thereof is known to play a role. Thus, another embodiment of the present disclosure relates to treating one or more diseases in which NRLP3 or a mutant thereof is known to play a role or reducing their severity.

[0178] Combined treatment method

[0179] The present disclosure provides combination therapies using the compounds as described in the present disclosure and other therapeutic agents. The term "combination therapy" as used in the present disclosure includes administering these agents in a sequential manner, i.e., where each therapeutic agent is administered at a different time, and administering these therapeutic agents, or at least two of the agents, substantially simultaneously. The order of each reagent, or substantially simultaneous administration, can be affected by any suitable route, including, but not limited to, oral route, intravenous route, intramuscular, subcutaneous route, and direct absorption through mucosal tissue. The agents can be administered by the same route or different routes. For example, the first agent can be administered orally, while the second agent is administered intravenously. In addition, the selected combination agent can be administered by intravenous injection, while the other agents in the combination can be administered orally. Alternatively, for example, two or more agents can be administered by intravenous or subcutaneous injection.

[0180] II. Examples

[0181] The present disclosure is further illustrated below with reference to examples. The description of specific exemplary embodiments of the present disclosure is for purposes of illustration and exemplification. These descriptions are not intended to limit the present disclosure to the precise forms disclosed, and it is obvious that many changes and variations can be made in accordance with the teachings of the present application specification. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present disclosure and its practical applications, so that those skilled in the art can implement and utilize various different exemplary embodiments of the present disclosure as well as various different selections and changes.

[0182] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods.

[0183] Unless otherwise specified, the materials, reagents, etc. used in the following examples are all commercially available.

[0184] Example 1: Synthesis of (R)-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0185]

[0186] Step 1: Dissolve phthalic anhydride (1a) (1.0 g, 6.75 mmol) and N2H4·H2O (1.01 g, 20.25 mmol) in acetic acid (20 mL), heat to 120 °C and reflux overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution to dryness, add water (20 ml) to disperse the solid, filter, dry the filter cake by suction and then dry it under vacuum overnight to obtain the target compound 1b (2,3-dihydrophthalazine-1,4-dione) (0.75 g, yield: 68.51%, LCMS m / z = 163.2 [M+1] + ).

[0187] Step 2: Dissolve compound (1b) (0.20 g, 1.23 mmol) in POCl3 (2 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the oily crude product dropwise to ice water (20 ml), stir well, adjust the pH = 8 with 2N sodium hydroxide aqueous solution, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound 1c (1,4-dichlorophthalazine) (0.21 g, yield: 85.54%, LCMS m / z = 199.2 [M+1] + ).

[0188] Step 3: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-1-methylpiperidin-3-amine (0.126 g, 1.11 mmol), and Na2CO3 (0.21 g, 2.01 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (1d) ((R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine) (0.06 g, yield: 21.57%, LCMS m / z = 277.2 [M+1] + ).

[0189] Step 4: Add compound (1d) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.045 g, 0.217 mmol), sodium carbonate (0.04 g, 0.36 mmol) and Pd(dppf)Cl2 (15.00 mg, 0.02 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1), displace with nitrogen 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain compound 1 ((R)-2-(4-((1-methylpiperidin-3-yl)amine)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.01 g, yield: 13.76%, LCMS m / z = 403.2 [M+1] + )。

[0190] Example 2: Synthesis of (R)-2-(4-((1-methylpiperidin-3-yl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)-5-(trifluoromethyl)phenol

[0191]

[0192] Step 1: Dissolve compound 4,5,6,7-tetrahydroisobenzofuran-1,3-dione (2a) (1.0 g, 6.57 mmol) and N2H4·H2O (0.98 g, 19.72 mmol) in acetic acid (20 mL), and heat to 120 °C for reflux overnight. Determine that the reaction is complete by TLC, directly concentrate the reaction solution to dryness, add water (20 ml) to disperse the solid, filter, and vacuum dry the filter cake overnight to obtain the target compound 2b (2,3,5,6,7,8-hexahydrophthalazine-1,4-dione) (0.60 g, yield: 54.93%, LCMS m / z = 167.2 [M+1] + )。

[0193] Step 2: Dissolve compound (2b) (0.20 g, 1.20 mmol) in POCl3 (2 mL), heat to 100 °C and react overnight. Determine the completion of the reaction by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil to ice water (20 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound (2c) (1,4-dichloro-5,6,7,8-tetrahydrophthalazine) (0.15 g, yield: 61.38%, LCMS m / z = 204.2 [M+1] + ).

[0194] Step 3: Dissolve compound (2c) (0.15 g, 0.738 mmol), (R)-1-methylpiperidin-3-amine (0.093 g, 0.81 mmol), and Na2CO3 (0.16 g, 1.48 mmol) in dry DMF (2 mL). Heat this mixed system in a sealed tube to 120 °C and react overnight. Determine the complete conversion of the starting materials by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (2d) ((R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine) (0.05 g, yield: 24.11%, LCMS m / z = 281.2 [M+1] + ).

[0195] Step 4: Add compound (2d) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.044 g, 0.214 mmol), sodium carbonate (0.037 g, 0.36 mmol) and Pd(dppf)Cl2 (15.00 mg, 0.02 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 2 ((R)-2-(4-((1-methylpiperidin-3-yl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.011 g, yield: 15.20%, LCMS m / z = 407.2 [M+1] + )

[0196] Example 3: Synthesis of (R)-2-(7-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0197]

[0198] Step 1: Dissolve compound 4-methylphthalic anhydride (3a) (2.0 g, 12.33 mmol) and N2H4·H2O (1.85 g, 37.00 mmol) in acetic acid (30 mL), and heat to 120 °C for reflux overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution to dryness, add water (20 ml) to disperse the solid, filter, and dry the filter cake under vacuum overnight to obtain the target compound 3b (6-methyl-2,3-dihydrophthalazine-1,4-dione) (1.51 g, yield: 69.03%, LCMS m / z = 177.2 [M+1] + )

[0199] Step 2: Dissolve compound (3b) (1.51 g, 8.57 mmol) in POCl3 (10 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil to ice water (50 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 10:1) to obtain the target compound (3c) (1,4-dichloro-6-methylphthalazine) (1.45 g, yield: 79.40%, LCMS m / z = 213.2 [M+1] + ).

[0200] Step 3: Dissolve compound (3c) (0.20 g, 0.94 mmol), (R)-1-methylpiperidin-3-amine (0.12 g, 1.04 mmol), and Na2CO3 (0.20 g, 1.89 mmol) in dry DMF (2 mL). Heat this mixed system in a sealed tube to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of the target compounds (3d) and (4d) (0.08 g, yield: 29.31%, LCMS m / z = 291.2 [M+1] + ).

[0201] Step 4: Add the mixture of compounds (3d) and (4d) (0.08 g, 0.27 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.074 g, 0.357 mmol), sodium carbonate (0.058 g, 0.55 mmol), and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace with nitrogen 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by PTLC (DCM:CH3OH = 10:1) to obtain the target compound 3 ((R)-2-(7-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.011 g, yield: 8.73%, LCMS m / z = 417.2 [M+1]+ )。

[0202] Example 4: Synthesis of (R)-2-(6-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0203]

[0204] Step 1: Dissolve compound (3c) (0.20 g, 0.94 mmol), (R)-1-methylpiperidin-3-amine (0.12 g, 1.04 mmol), and Na2CO3 (0.20 g, 1.89 mmol) in dry DMF (2 mL). Heat this mixed system in a sealed tube at 120 °C overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure from the filtrate. Separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of the target compound (4d) and (3d) (0.08 g, yield: 29.31%, LCMS m / z = 291.2 [M+1] + )。

[0205] Step 2: Add a mixture of compound (3d) and (4d) (0.08 g, 0.27 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.074 g, 0.357 mmol), sodium carbonate (0.058 g, 0.55 mmol), and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace with nitrogen 3 times and heat at 110 °C for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure from the filtrate. Separate the residue by PTLC (DCM:CH3OH = 10:1) to obtain the target compound 4 ((R)-2-(6-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.008 g, yield: 6.98%, LCMS m / z = 417.2 [M+1] + )。

[0206] Example 5: Synthesis of (R)-2-(7-methoxy-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0207]

[0208] Step 1: Dissolve compound 4-methoxyphthalic anhydride (5a) (2.0 g, 11.23 mmol) and N2H4·H2O (1.69 g, 33.68 mmol) in acetic acid (30 mL), heat to 120 °C and reflux overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution to dryness, add water (20 ml) to disperse the solid, filter, and dry the filter cake under vacuum overnight to obtain the target compound 5b (6-methoxy-2,3-dihydrophthalazine-1,4-dione) (1.42 g, yield: 65.82%, LCMS m / z = 193.2 [M+1] + )

[0209] Step 2: Dissolve compound (5b) (1.42 g, 7.39 mmol) in POCl3 (10 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil product dropwise to ice water (50 ml), stir well, adjust the pH = 8 with 2N sodium hydroxide aqueous solution, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 10:1) to obtain the target compound (5c) (1,4-dichloro-6-methoxyphthalazine) (1.20 g, yield: 70.90%, LCMS m / z = 229.2 [M+1] + )

[0210] Step 3: Dissolve compound (5c) (0.20 g, 0.87 mmol), (R)-1-methylpiperidin-3-amine (0.11 g, 0.96 mmol), and Na2CO3 (0.20 g, 1.89 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of the target compounds (5d) and (6d) (0.09 g, yield: 33.60%, LCMS m / z = 307.2 [M+1] + )

[0211] Step 4: Add the mixture of compound (5d) and (6d) (0.09 g, 0.29 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (78.53 mg, 0.38 mmol), sodium carbonate (62.18 mg, 0.58 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat the mixture to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by PTLC (DCM:CH3OH = 10:1) to obtain the target compound 5 ((R)-2-(7-methoxy-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.01 g, yield: 7.88%, LCMS m / z = 433.2 [M+1] + )。

[0212] Example 6: Synthesis of (R)-2-(6-methoxy-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0213]

[0214] Step 1: Dissolve compound (5c) (0.20 g, 0.87 mmol), (R)-1-methylpiperidin-3-amine (0.11 g, 0.96 mmol), and Na2CO3 (0.20 g, 1.89 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of the target compounds (6d) and (5d) (0.09 g, yield: 33.60%, LCMS m / z = 307.2 [M+1] + )。

[0215] Step 2: Add the mixture of compound (5d) and (6d) (0.09 g, 0.29 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (78.53 mg, 0.38 mmol), sodium carbonate (62.18 mg, 0.58 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat the mixture to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by PTLC (DCM:CH3OH = 10:1) to obtain the target compound 6 ((R)-2-(6-methoxy-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.008 g, yield: 6.31%, LCMS m / z = 433.2 [M+1] + )。

[0216] Example 7: Synthesis of (R)-2-(4-((1-phenylethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0217]

[0218] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-1-methylbenzylamine (0.15 g, 1.21 mmol), and Na2CO3 (0.21 g, 2.01 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (7a) ((R)-4-chloro-N-(1-phenylethyl)phthalazin-1-amine (7a)) (0.14 g, yield: 49.10%, LCMS m / z = 284.2 [M+1] + )。

[0219] Step 2: Add compound (7a) (0.05 g, 0.17 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (39.91 mg, 0.19 mmol), sodium carbonate (37.35 mg, 0.35 mmol) and Pd(dppf)Cl2 (10.24 mg, 0.014 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen for 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 7 ((R)-2-(4-((1-phenylethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.02 g, yield: 27.72%, LCMS m / z = 410.2 [M+1] + )。

[0220] Example 8: Synthesis of (S)-2-(4-((1-phenylethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0221]

[0222] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (S)-1-methylbenzylamine (0.15 g, 1.21 mmol), and Na2CO3 (0.21 g, 2.01 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (8a) ((S)-4-chloro-N-(1-phenylethyl)phthalazin-1-amine) (0.14 g, yield: 49.10%, LCMS m / z = 284.2 [M+1] + )。

[0223] Step 2: Add compound (8a) (0.05 g, 0.17 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (39.91 mg, 0.19 mmol), sodium carbonate (37.35 mg, 0.35 mmol) and Pd(dppf)Cl2 (10.24 mg, 0.014 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 8 ((S)-2-(4-((1-phenylethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.02 g, yield: 27.72%, LCMS m / z = 410.2 [M+1] + )。

[0224] Example 9: Synthesis of (R)-2-(4-((1-(4-fluorophenyl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0225]

[0226] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-1-(4-fluorophenyl)ethan-1-amine (0.16 g, 1.20 mmol), and Na2CO3 (0.21 g, 2.01 mmol) in dry DMF (2 mL), place this mixed system in a sealed tube and heat to 120 °C to react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (9a) ((R)-4-chloro-N-(1-(4-fluorophenyl)ethyl)phthalazin-1-amine) (0.12 g, yield: 39.58%, LCMS m / z = 302.2 [M+1] + )。

[0227] Step 2: Add compound (9a) (0.05 g, 0.16 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (37.53 mg, 0.18 mmol), sodium carbonate (35.12 mg, 0.33 mmol) and Pd(dppf)Cl2 (10.24 mg, 0.014 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 9 (((R)-2-(4-((1-(4-fluorophenyl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.02 g, yield: 28.24%, LCMS m / z = 428.2 [M+1] + ).

[0228] Example 10: Synthesis of (S)-2-(4-((1-(4-fluorophenyl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0229]

[0230] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (S)-1-(4-fluorophenyl)ethan-1-amine (0.16 g, 1.20 mmol), and Na2CO3 (0.21 g, 2.01 mmol) in dry DMF (2 mL), place this mixed system in a sealed tube, heat to 120 °C and react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (10a) ((S)-4-chloro-N-(1-(4-fluorophenyl)ethyl)phthalazin-1-amine) (0.12 g, yield: 39.58%, LCMS m / z = 302.2 [M+1] + ).

[0231] Step 2: Add compound (10a) (0.05 g, 0.16 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (37.53 mg, 0.18 mmol), sodium carbonate (35.12 mg, 0.33 mmol) and Pd(dppf)Cl2 (10.24 mg, 0.014 mol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 10 (((S)-2-(4-((1-(4-fluorophenyl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.015 g, yield: 21.18%, LCMS m / z = 428.2 [M+1] + ).

[0232] Example 11: Synthesis of 2-(4-(butylamino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0233]

[0234] Step 1: Dissolve compound (1c) (0.10 g, 0.50 mmol), n-butylamine (0.036 g, 0.50 mmol), and Na2CO3 (0.11 g, 1.00 mmol) in dry DMF (2 mL), place this mixed system in a sealed tube, heat to 120 °C and react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (11a) (n-butyl-4-chlorophthalazin-1-amine) (0.05 g, yield: 42.22%, LCMS m / z = 236.2 [M+1] + ).

[0235] Step 2: Add compound (11a) (0.05 g, 0.21 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (52.42 mg, 0.25 mmol), sodium carbonate (44.96 mg, 0.42 mmol) and Pd(dppf)Cl2 (10.24 mg, 0.014 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat the mixture to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 11 ((2-(4-(butylamino)phthalazin-1-yl)-5-(trifluoromethyl)) (0.025 g, yield: 32.61%, LCMS m / z = 362.2 [M+1] + )。

[0236] Example 12: (R)-2-(4-((1-(pyridin-2-yl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0237]

[0238] Step 1: Dissolve compound (1c) (0.10 g, 0.50 mmol), (R)-1-(pyridin-2-yl)ethan-1-amine (67.52 mg, 0.55 mmol), and Na2CO3 (0.11 g, 1.00 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (12a) ((R)-4-chloro-N-(1-(pyridin-2-yl)ethyl)phthalazin-1-amine) (0.06 g, yield: 41.94%, LCMS m / z = 285.2 [M+1] + )。

[0239] Step 2: Add compound (12a) (0.06 g, 0.21 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (52.07 mg, 0.25 mmol), sodium carbonate (44.67 mg, 0.42 mmol) and Pd(dppf)Cl2 (15.36 mg, 0.021 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen for 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 12 ((R)-2-(4-((1-(pyridin-2-yl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.025 g, yield: 28.91%, LCMS m / z = 411.2 [M+1] + )。

[0240] Example 13: Synthesis of (S)-2-(4-((1-(pyridin-2-yl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0241]

[0242] Step 1: Dissolve compound (1c) (0.10 g, 0.50 mmol), (S)-1-(pyridin-2-yl)ethan-1-amine (67.52 mg, 0.55 mmol), and Na2CO3 (0.11 g, 1.00 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound (13a) ((S)-4-chloro-N-(1-(pyridin-2-yl)ethyl)phthalazin-1-amine) (0.05 g, yield: 34.95%, LCMS m / z = 285.2 [M+1] + )。

[0243] Step 2: Add compound (13a) (0.05 g, 0.17 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (43.39 mg, 0.21 mmol), sodium carbonate (38.22 mg, 0.35 mmol) and Pd(dppf)Cl2 (15.36 mg, 0.021 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 3:1) to obtain the target compound 13 ((S)-2-(4-((1-(pyridin-2-yl)ethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.02 g, yield: 27.75%, LCMS m / z = 411.2 [M+1] + )。

[0244] Example 14: Synthesis of 2-(5-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol

[0245]

[0246] Step 1: Dissolve compound furano[3,4-b]pyridine-5,7-dione (14a) (3.0 g, 20.12 mmol) and N2H4·H2O (3.02 g, 60.36 mmol) in acetic acid (50 mL), heat to 120 °C and reflux overnight. Determine that the reaction is complete by TLC, directly concentrate the reaction solution to dryness, disperse the solid with water (50 ml), filter, dry the filter cake under vacuum overnight to obtain the target compound (14b) (6,7-dihydropyrido[2,3-d]pyridazine-5,8-dione) (2.30 g, yield: 70.07%, LCMS m / z = 164.2 [M+1] + )。

[0247] Step 2: Dissolve compound (14b) (2.30 g, 14.10 mmol) in POCl3 (20 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil to ice water (50 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 10:1) to obtain the target compound (14c) (5,8-dichloropyrido[2,3-d]pyridazine) (1.25 g, yield: 44.33%, LCMS m / z = 200.2 [M+1] + ).

[0248] Step 3: Dissolve compound (14c) (0.20 g, 1.00 mmol), (R)-1-methylpiperidin-3-amine (0.14 g, 1.20 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube, heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (14d) ((3R)-N-(8-chloropyrido[2,3-d]pyridazin-5-yl)-1-methylpiperidin-3-amine) (0.13 g, yield: 46.80%, LCMS m / z = 278.2 [M+1] + ).

[0249] Step 4: Add compound (14d) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (44.00 mg, 0.22 mmol), sodium carbonate (29.15 mg, 0.27 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 14 (2-(5-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol) (0.022 g, yield: 30.30%, LCMS m / z = 404.4 [M+1] + )。

[0250] Example 15: Synthesis of 2-(8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol

[0251]

[0252] Step 1: Dissolve compound (14c) (0.20 g, 1.00 mmol), (R)-1-methylpiperidin-3-amine (0.14 g, 1.20 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube and heat to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (15d) ((3R)-N-(5-chloropyrido[2,3-d]pyridazin-8-yl)-1-methylpiperidin-3-amine) (0.04 g, yield: 14.40%, LCMS m / z = 278.2 [M+1] + )。

[0253] Step 2: Add compound (15d) (0.04 g, 0.14 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35.00 mg, 0.17 mmol), sodium carbonate (22.10 mg, 0.21 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 15 (2-(8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol) (0.015 g, yield: 26.56%, LCMS m / z = 404.4 [M+1] + ).

[0254] Example 16: Synthesis of 2-(7-fluoro-4-{[(3R)-1-methylpiperidin-3-yl]amino}phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0255]

[0256] Step 1: Dissolve compound 5-fluoroisobenzofuran-1,3-dione (16a) (5.0 g, 30.10 mmol) and N2H4·H2O (7.53 g, 150.50 mmol) in acetic acid (50 mL), heat to 120 °C and reflux overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution to dryness, disperse the solid with water (50 ml), filter, dry the filter cake by suction and then dry it under vacuum overnight to obtain the target compound (16b) (6-fluoro-2,3-dihydrophthalazine-1,4-dione) (5.40 g, yield: 99.59%, LCMS m / z = 181.1 [M+1] + ).

[0257] Step 2: Dissolve compound (16b) (5.40 g, 29.98 mmol) in POCl3 (30 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil to ice water (50 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (50 mL × 3), combine the organic phases, wash with saturated brine (50 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 5:1) to obtain the target compound (16c) (1,4-dichloro-6-fluorophthalazine) (4.83 g, yield: 74.23%, LCMS m / z = 217.0 [M+1] + )

[0258] Step 3: Dissolve compound (16c) (0.20 g, 0.92 mmol), (R)-1-methylpiperidin-3-amine (0.14 g, 1.20 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Heat this mixed system in a sealed tube to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (16d) ((R)-4-chloro-6-fluoro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine) (0.05 g, yield: 18.41%, LCMS m / z = 295.2 [M+1] + )

[0259] Step 4: Add compound (16d) (0.05 g, 0.17 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (44.00 mg, 0.22 mmol), sodium carbonate (29.15 mg, 0.27 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1). Replace the air with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 16 ((R)-2-(7-fluoro-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.01 g, yield: 14.02%, LCMS m / z = 421.4 [M+1] + )。

[0260] Example 17: Synthesis of (R)-2-(1-((1-methylpiperidin-3-yl)amino)pyrido[3,4-d]pyridazin-4-yl)-5-(trifluoromethyl)phenol

[0261]

[0262] Step 1: Dissolve compound furan[3,4-c]pyridine-1,3-dione (17a) (3.0 g, 20.12 mmol) and N2H4.H2O (3.02 g, 60.36 mmol) in acetic acid (50 mL), heat to 120 °C and reflux overnight. Determine that the reaction is complete by TLC. Concentrate the reaction solution directly to dryness, disperse the solid with water (50 ml), filter, dry the filter cake under vacuum overnight to obtain the target compound (17b) (2,3-dihydropyrido[3,4-d]pyridazine-1,4-dione) (2.10 g, yield: 63.97%, LCMS m / z = 164.2 [M+1] + )。

[0263] Step 2: Dissolve compound (17b) (2.10 g, 12.87 mmol) in POCl3 (20 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil to ice water (50 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 10:1) to obtain the target compound (17c) (1,4-dichloropyrido[3,4-d]pyridazine) (0.92 g, yield: 32.63%, LCMS m / z = 200.2 [M+1] + ).

[0264] Step 3: Dissolve compound (17c) (0.20 g, 1.00 mmol), (R)-1-methylpiperidin-3-amine (0.14 g, 1.20 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Heat this mixed system in a sealed tube to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (17d) ((R)-4-chloro-N-(1-methylpiperidin-3-yl)pyrido[3,4-d]pyridazin-1-amine) (0.11 g, yield: 39.60%, LCMS m / z = 278.2 [M+1] + ).

[0265] Step 4: Add compound (17d) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (44.00 mg, 0.22 mmol), sodium carbonate (29.15 mg, 0.27 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1), displace with nitrogen 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 17 ((R)-2-(1-((1-methylpiperidin-3-yl)amino)pyrido[3,4-d]pyridazin-4-yl)-5-(trifluoromethyl)phenol) (0.01 g, yield: 13.77%, LCMS m / z = 404.4 [M+1] + )。

[0266] Example 18: Synthesis of 2-(4-((trans)-4-(dimethylamino)cyclohexyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0267]

[0268] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (trans)-N 1 ,N 1 -dimethylcyclohexane-1,4-diamine (0.14 g, 1.00 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Heat this mixed system in a sealed tube to 120 °C and react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (18b) (trans)-N 1 -(4-chlorophthalazin-1-yl)-N 4 ,N 4 -dimethylcyclohexane-1,4-diamine (0.04 g, yield: 13.15%, LCMS m / z = 305.2 [M+1] + )。

[0269] Step 2: Add compound (18b) (0.04 g, 0.13 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35.00 mg, 0.17 mmol), sodium carbonate (22.10 mg, 0.21 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure for the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 18 2-(4-((trans)-4-(dimethylamino)cyclohexyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.012 g, yield: 21.46%, LCMS m / z = 431.5 [M+1] + )。

[0270] Example 19: Synthesis of 2-(4-((cis)-4-(dimethylamino)cyclohexyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0271]

[0272] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (cis)-N 1 ,N 1 -dimethylcyclohexane-1,4-diamine (0.14 g, 1.00 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL), place this mixed system in a sealed tube and heat to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted, add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure for the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (19b) (cis)-N 1 -(4-chlorophthalazin-1-yl)-N 4 ,N 4 -dimethylcyclohexane-1,4-diamine (0.05 g, yield: 16.45%, LCMS m / z = 305.2 [M+1] + )。

[0273] Step 2: Add compound (19b) (0.05 g, 0.13 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35.00 mg, 0.17 mmol), sodium carbonate (22.10 mg, 0.21 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 19 2-(4-((cis)-4-(dimethylamino)cyclohexyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.020 g, yield: 35.78%, LCMS m / z = 431.5 [M+1] + )

[0274] Example 20: Synthesis of 2-(4-((2-hydroxy-2-methylpropyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0275]

[0276] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), 1-amino-2-methylpropan-2-ol (0.14 g, 1.00 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube and heat to 120 °C to react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (20b) 1-((4-chlorophthalazin-1-yl)amino)-2-methylpropan-2-ol (0.05 g, yield: 19.92%, LCMS m / z = 252.1 [M+1] + )

[0277] Step 2: Add compound (20b) (0.05 g, 0.20 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (50.00 mg, 0.24 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 20 2-(4-((2-hydroxy-2-methylpropyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.01 g, yield: 11.60%, LCMS m / z = 378.4 [M+1] + )

[0278] Example 21: Synthesis of 2-(4-((2-morpholinoethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0279]

[0280] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), 2-morpholinoethan-1-amine (0.12 g, 1.00 mmol) and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube and heat to 120 °C to react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (21b) 4-chloro-N-(2-morpholinoethyl)phthalazin-1-amine (0.05 g, yield: 17.12%, LCMS m / z = 293.1 [M+1] + )

[0281] Step 2: Add compound (21b) (0.05 g, 0.17 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (42.00 mg, 0.20 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 21 2-(4-((2-morpholinoethyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.01 g, yield: 11.60%, LCMS m / z = 419.4 [M+1] + )。

[0282] Example 22: Synthesis of (R)-2-(4-(piperidin-3-ylamino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0283]

[0284] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-tert-butyl 3-aminopiperidine-1-carboxylate (0.20 g, 1.00 mmol) and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (10 mL). Place this mixed system in a 50 mL single-necked flask and heat to 120 °C to react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (EA:PE = 1:1) to obtain the target compound (22a) (tert-butyl (R)-3-((4-chlorophthalazin-1-yl)amino)piperidine-1-carboxylate) (0.22 g, yield: 60.34%, LCMS m / z = 363.2 [M+1] + )。

[0285] Step 2: Dissolve compound (22a) (0.22 g, 0.61 mmol) in dichloromethane (10 mL), add it to trifluoroacetic acid (2 mL), and react at room temperature overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution. After removing trifluoroacetic acid, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure from the filtrate to obtain the target compound (22b) ((R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine) (0.11 g, yield: 69.05%, LCMS m / z = 263.2 [M+1] + ).

[0286] Step 3: Add compound (22b) (0.05 g, 0.19 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (44.00 mg, 0.22 mmol), sodium carbonate (29.15 mg, 0.27 mmol), and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1), displace with nitrogen 3 times, and heat to 110 °C for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure from the filtrate. Separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 22 ((R)-2-(4-(piperidin-3-ylamino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.03 g, yield: 40.59%, LCMS m / z = 389.2 [M+1] + ).

[0287] Example 23: Synthesis of (R)-2-(4-((1-ethylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0288]

[0289] Step 1: Dissolve compound (22) (0.02 g, 0.051 mmol) and acetaldehyde tetrahydrofuran solution (0.20 mL, 5.0 M) in dry tetrahydrofuran (5 mL). Add two drops of acetic acid. After stirring the mixture at room temperature for 1 hour in a 50 mL single-necked flask, add NaBH(OAc)3 and continue the reaction for 3 hours. When the reaction is complete by TLC, add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure from the filtrate. The residue is separated by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (23) ((R)-2-(4-((1-ethylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.01 g, yield: 46.63%, LCMS m / z = 417.4 [M+1] + )。

[0290] Example 24: (R)-2-(4-((1-methylpyrrolidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0291]

[0292] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-1-methylpyrrolidin-3-amine (0.11 g, 1.1 mmol), and Na2CO3 (0.21 g, 2.0 mmol) in dry DMF (2 mL). Heat the mixture in a sealed tube at 120 °C overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure from the filtrate. The residue is separated by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (24a) ((R)-4-chloro-N-(1-methylpyrrolidin-3-yl)phthalazin-1-amine) (0.06 g, yield: 21.6%, LCMS m / z = 263.4 [M+1] + )。

[0293] Step 2: At room temperature, add compound (24a) (27 mg, 0.10 mmol), 2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.045 g, 0.217 mmol), (79 mg, 0.36 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly separate by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound 24 ((R)-2-(4-((1-methylpyrrolidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (30 mg, yield: 71.24%, LCMS m / z = 389.4 [M+1] + )

[0294] Example 25: (R)-2-(4-((1-ethylpyrrolidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0295]

[0296] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-1-ethylpyrrolidin-3-amine (0.14 g, 1.1 mmol), and Na2CO3 (0.21 g, 2.0 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C to react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (25a) ((R)-4-chloro-N-(1-ethylpyrrolidin-3-yl)phthalazin-1-amine) (0.10 g, yield: 36.23%, LCMS m / z = 277.4 [M+1] + )。

[0297] Step 2: At room temperature, add compound (24a) (28 mg, 0.10 mmol), 2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.045 g, 0.217 mmol), (79 mg, 0.36 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into a 5 mL mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly separate it by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound 25 ((R)-2-(4-((1-ethylpyrrolidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (20 mg, yield: 49.75%, LCMS m / z = 403.4 [M+1] + )

[0298] Example 27: (R)-2-(4-(((1-ethylpyrrolidin-2-yl)methyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0299]

[0300] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-(1-ethylpyrrolidin-2-yl)methylamine (0.11 g, 1.1 mmol), and Na2CO3 (0.21 g, 2.0 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C to react overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (27a) ((R)-4-chloro-N-((1-ethylpyrrolidin-2-yl)methyl)phthalazin-1-amine) (0.08 g, yield: 27.5%, LCMS m / z = 291.4 [M+1] + )。

[0301] Step 2: At room temperature, add compound (27a) (30 mg, 0.10 mmol), 2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.045 g, 0.217 mmol), (79 mg, 0.36 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 27 ((R)-2-(4-(((1-ethylpyrrolidin-2-yl)methyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (10 mg, yield: 23.98%, LCMS m / z = 417.4 [M+1] + ) Example 29: Synthesis of (R)-2-(4-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0302]

[0303] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (R)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-amine (0.13 g, 1.00 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube and heat to 120 °C for overnight reaction. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and perform column chromatography separation on the residue (DCM:CH3OH = 10:1) to obtain the target compound (29b) (R)-4-chloro-N-(6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)phthalazin-1-amine (0.06 g, yield: 20.27%, LCMS m / z = 297.8 [M+1] + )

[0304] Step 2: Add compound (29b) (0.06 g, 0.20 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (50.00 mg, 0.24 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat the mixture to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 29 (R)-2-(4-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.01 g, yield: 11.84%, LCMS m / z = 423.5 [M+1] + )

[0305] Example 30: Synthesis of (S)-2-(4-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0306]

[0307] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), (S)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-amine (0.13 g, 1.00 mmol) and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution into water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (30b) (S)-4-chloro-N-(6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)phthalazin-1-amine (0.05 g, yield: 16.83%, LCMS m / z = 297.8 [M+1] + )

[0308] Step 2: Add compound (30b) (0.06 g, 0.17 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (42.00 mg, 0.20 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 30 (R)-2-(4-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.01 g, yield: 13.94%, LCMS m / z = 423.5 [M+1] + ).

[0309] Example 31: (R)-2-(4-((1-Methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol

[0310]

[0311] Step 1: At room temperature, add compound (1d) (50 mg, 0.18 mmol), (2-hydroxyphenyl)boronic acid (50 mg, 0.36 mmol), sodium carbonate (60 mg, 0.57 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly separate by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound 31 ((R)-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol) (20 mg, yield: 33.23%, LCMS m / z = 335.4 [M+1] + )

[0312] Example 32: Synthesis of (R)-2-(4-((1-Ethylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0313]

[0314] Step 1: Dissolve compound (22b) (0.01 g, 0.038 mmol) and triethylamine (77.03 mg, 0.76 mmol) in dichloromethane (5 mL). Dropwise add acetyl chloride (29.88 mg, 0.038 mmol) and react at room temperature for 0.5 h. When the reaction is complete by TLC, add the reaction solution to water (20 ml), stir well, extract with DCM (20 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (EA:PE = 1:3) to obtain the target compound (32a) ((R)-1-(3-((4-chlorophthalazin-1-yl)amino)piperidin-1-yl)ethan-1-one) (0.06 g, yield: 51.72%, LCMS m / z = 305.2 [M+1] + ).

[0315] Step 2: Add compound (32a) (0.05 g, 0.16 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (44.00 mg, 0.22 mmol), sodium carbonate (29.15 mg, 0.27 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times and heat to 110 °C for reaction for 3 h. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 32 ((R)-1-(3-((4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1-yl)amino)piperidin-1-yl)ethan-1-one) (0.02 g, yield: 28.32%, LCMS m / z = 431.2 [M+1] + ).

[0316] Example 33: Synthesis of Isomer 1 and Isomer 2 of 2-(4-((-3-hydroxycyclohexyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0317]

[0318] Step 1: Dissolve compound (1c) (0.28 g, 1.39 mmol), 3-aminocyclohexanol (0.16 g, 1.39 mmol), and Na2CO3 (0.44 g, 4.17 mmol) in dry DMAc (2.5 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (33a) 3-((4-chlorophthalazin-1-yl)amino)cyclohexan-1-ol (0.35 g, yield: 90.66%, LCMS m / z = 278.2 [M+1] + ).

[0319] Step 2: Add compound (33a) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.048 g, 0.23 mmol), sodium carbonate (0.057 g, 0.54 mmol), and Pd(dppf)Cl2 (15.00 mg, 0.02 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen 3 times, and heat to 110 °C for reaction for 16 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 20:1) to obtain isomer 1 (compound 33) of compound (2-(4-((-3-hydroxycyclohexyl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (developing agent DCM:CH3OH = 10:1, Rf = 0.2, 15 mg, yield: 20.66%, LC-MS m / z = 404.5 [M+1] + ) and isomer 2 (compound 34) (developing agent DCM:CH3OH = 10:1, Rf = 0.3, 15 mg, yield: 20.66%, LCMS m / z = 404.2 [M+1] + ).

[0320] Example 35: Synthesis of 2-(4-((1-methylpiperidin-4-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0321]

[0322] Step 1: Dissolve compound (1c) (0.20 g, 1.00 mmol), 1-methylpiperidin-4-amine (0.12 g, 1.00 mmol), and Na2CO3 (0.21 g, 1.99 mmol) in dry DMF (3 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (35b) 4-chloro-N-(1-methylpiperidin-4-yl)phthalazin-1-amine (0.06 g, yield: 21.73%, LCMS m / z = 277.2 [M+1] + ).

[0323] Step 2: Add compound (35b) (0.03 g, 0.21 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (25.00 mg, 0.24 mmol), sodium carbonate (21.20 mg, 0.20 mmol), and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 35 2-(4-((1-methylpiperidin-4-yl)amino)phthalazin-1-yl)-5-(trifluoromethyl)phenol (0.02 g, yield: 23.69%, LCMS m / z = 403.4 [M+1] + ).

[0324] Example 36: Synthesis of (R)-2-(1-methyl-7-((1-methylpiperidin-3-yl)amino)-1H-imidazo[4,5-d]pyridazin-4-yl)-5-(trifluoromethyl)phenol

[0325]

[0326] Step 1: Dissolve dimethyl 1H-imidazole-4,5-dicarboxylate (36a) (3.0 g, 16.29 mmol), methyl iodide (3.47 g, 24.43 mmol), and potassium carbonate (3.38 g, 24.43 mmol) in DMF (20 mL), and heat the solution to 60 °C for 3 hours. Determine the completion of the reaction by TLC. Add the reaction solution to water (50 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (36b) (dimethyl 1-methyl-1H-imidazole-4,5-dicarboxylate) (0.80 g, yield: 24.78%, LCMS m / z = 199.2 [M+1] + )

[0327] Step 2: Dissolve dimethyl 1-methyl-1H-imidazole-4,5-dicarboxylate (36b) (0.8 g, 4.04 mmol) and N2H4·H2O (0.61 g, 12.12 mmol) in acetic acid (5 mL), and heat the solution to 120 °C for reflux overnight. Determine the completion of the reaction by TLC. Concentrate the reaction solution directly to dryness, add water (30 mL) to disperse the solid, filter, and dry the filter cake under vacuum overnight to obtain the target compound (36c) (1-methyl-5,6-dihydro-1H-imidazo[4,5-d]pyridazine-4,7-dione) (0.40 g, yield: 59.59%, LCMS m / z = 167.2 [M+1] + )

[0328] Step 3: Dissolve compound (36c) (0.40 g, 2.41 mmol) in POCl3 (10 mL), and heat the solution to 100 °C for reaction overnight. Determine the completion of the reaction by TLC. Concentrate the reaction solution directly, remove POCl3, and slowly add the crude oil to ice water (50 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by column chromatography (PE:EA = 5:1) to obtain the target compound (36d) (4,7-dichloro-1-methyl-1H-imidazo[4,5-d]pyridazine) (0.40 g, yield: 81.75%, LCMS m / z = 203.2 [M+1] + )

[0329] Step 4: Dissolve compound (36d) (0.40 g, 1.97 mmol), (R)-1-methylpiperidin-3-amine (0.27 g, 2.36 mmol), and Na2CO3 (0.21 g, 1.97 mmol) in dry DMF (5 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (36e) ((R)-4-chloro-1-methyl-N-(1-methylpiperidin-3-yl)-1H-imidazo[4,5-d]pyridazin-7-amine) (0.05 g, yield: 9.04%, LCMS m / z = 281.2 [M+1] + ).

[0330] Step 5: Add compound (36e) (0.04 g, 0.14 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35.00 mg, 0.17 mmol), sodium carbonate (22.10 mg, 0.22 mmol), and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace with nitrogen 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 36 ((R)-2-(1-methyl-7-((1-methylpiperidin-3-yl)amino)-1H-imidazo[4,5-d]pyridazin-4-yl)-5-(trifluoromethyl)phenol) (0.020 g, yield: 35.15%, LCMS m / z = 407.4 [M+1] + ).

[0331] Example 37: Synthesis of (R)-2-(1-methyl-4-((1-methylpiperidin-3-yl)amino)-1H-imidazo[4,5-d]pyridazin-7-yl)-5-(trifluoromethyl)phenol

[0332]

[0333] Step 1: Dissolve compound (36d) (0.40 g, 1.97 mmol), (R)-1-methylpiperidin-3-amine (0.27 g, 2.36 mmol), and Na2CO3 (0.21 g, 1.97 mmol) in dry DMF (5 mL). Place this mixed system in a sealed tube and heat it to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL×3), combine the organic phases, wash with saturated brine (10 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (37a) ((R)-7-chloro-1-methyl-N-(1-methylpiperidin-3-yl)-1H-imidazo[4,5-d]pyridazin-4-amine) (0.10 g, yield: 18.08%, LCMS m / z = 281.2 [M+1] + ).

[0334] Step 2: Add compound (37a) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (44.00 mg, 0.22 mmol), sodium carbonate (22.10 mg, 0.22 mmol), and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace with nitrogen 3 times and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 37 ((R)-2-(1-methyl-4-((1-methylpiperidin-3-yl)amino)-1H-imidazo[4,5-d]pyridazin-7-yl)-5-(trifluoromethyl)phenol) (0.030 g, yield: 41.04%, LCMS m / z = 407.4 [M+1] + ).

[0335] Example 38: Synthesis of (R)-2-(4-((1-methylpiperidin-3-yl)amino)-7-(trifluoromethyl)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0336]

[0337] Step 1: Dissolve 4-(trifluoromethyl)phthalic acid (38a) (2.0 g, 8.54 mmol) in 15 ml of thionyl chloride, add 2 drops of dry DMF, heat to reflux for 2 hours, concentrate this reaction solution to dryness, redissolve with toluene (20 mL) and then concentrate to dryness again. Dissolve the residue in acetic acid (30 mL), add N2H4.H2O (1.85 g, 37.00 mmol), heat to 120 °C and reflux overnight. Directly concentrate the reaction solution to dryness, add water (20 ml) to disperse the solid, filter, drain the filter cake and dry it under vacuum overnight to obtain the target compound 38b (6-(trifluoromethyl)-2,3-dihydrophthalazine-1,4-dione) (0.70 g, yield: 35.61%, LCMS m / z = 231.2 [M+1] + )。

[0338] Step 2: Dissolve compound (38b) (0.70 g, 3.04 mmol) in POCl3 (10 mL), heat to 100 °C and react overnight. Determine that the reaction is complete by TLC. Directly concentrate the reaction solution, remove POCl3, and slowly add the crude oil to ice water (50 ml), stir well, adjust the pH = 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 10:1) to obtain the target compound (38c) (1,4-dichloro-6-(trifluoromethyl)phthalazine) (0.52 g, yield: 64.02%, LCMS m / z = 267.2 [M+1] + )。

[0339] Step 3: Dissolve compound (38c) (0.22 g, 0.82 mmol), (R)-1-methylpiperidin-3-amine (0.12 g, 1.04 mmol), and Na2CO3 (0.20 g, 1.89 mmol) in dry DMF (2 mL), place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of the target compounds (38d) and (39d) (0.15 g, yield: 52.81%, LCMS m / z = 345.2 [M+1] + )。

[0340] Step 4: Add the mixture of compound (38d) and (39d) (0.10 g, 0.29 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.074 g, 0.357 mmol), sodium carbonate (0.058 g, 0.55 mmol) and Pd(dppf)Cl2 (20.00 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by PTLC (DCM:CH3OH = 10:1) to obtain the target compound 38 ((R)-2-(4-((1-methylpiperidin-3-yl)amino)-7-(trifluoromethyl)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.01 g, yield: 7.33%, LCMS m / z = 471.4 [M+1] + )。

[0341] Example 39: Synthesis of (R)-2-(4-((1-methylpiperidin-3-yl)amino)-6-(trifluoromethyl)phthalazin-1-yl)-5-(trifluoromethyl)phenol

[0342]

[0343] Step 1: Dissolve compound (38c) (0.22 g, 0.82 mmol), (R)-1-methylpiperidin-3-amine (0.12 g, 1.04 mmol), and Na2CO3 (0.20 g, 1.89 mmol) in dry DMF (2 mL). Place this mixed system in a sealed tube and heat to 120 °C for reaction overnight. Determine by TLC that the raw materials are completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of the target compounds (38d) and (39d) (0.15 g, yield: 52.81%, LCMS m / z = 345.2 [M+1] + )。

[0344] Step 2: Add the mixture of compound (38d) and (39d) (0.10 g, 0.29 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (0.074 g, 0.357 mmol), sodium carbonate (0.058 g, 0.55 mmol) and Pd(dppf)Cl2 (20.00 mg, 0.027 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat the mixture to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by PTLC (DCM:CH3OH = 10:1) to obtain the target compound 39 (R)-2-(4-((1-methylpiperidin-3-yl)amino)-6-(trifluoromethyl)phthalazin-1-yl)-5-(trifluoromethyl)phenol) (0.007 g, yield: 5.13%, LCMS m / z = 471.4 [M+1] + )

[0345] Example 40: Synthesis of (R)-5-methyl-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol

[0346]

[0347] Step: Add compound (1d) (0.05 g, 0.18 mmol), (2-hydroxy-4-methylphenyl)boronic acid (40.77 mg, 0.27 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat the mixture to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 40 (R)-5-methyl-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol (0.02 g, yield: 23.69%, LCMS m / z = 349.2 [M+1] + )

[0348] Example 41: Synthesis of (R)-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethoxy)phenol

[0349]

[0350] Procedure: Add compound (1d) (0.05 g, 0.18 mmol), (2-hydroxy-4-(trifluoromethoxy)phenyl)boronic acid (59.94 mg, 0.27 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1), displace with nitrogen for 3 times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 41 (R)-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-(trifluoromethoxy)phenol (0.02 g, yield: 26.52%, LCMS m / z = 419.2 [M+1] + )。

[0351] Example 43: (R)-(1-(2-Hydroxy-4-(trifluoromethyl)phenyl)-4-((1-methylpiperidin-3-yl)amino)phthalazin-6-yl)dimethylphosphine oxide

[0352]

[0353] Step 1: Dissolve compound (43a) (1.0 g, 4.44 mmol) and N2H4·H2O (2.5 g, 41.1 mmol, 80% wt) in acetic acid (20 mL), heat to 120 °C and reflux overnight. Determine that the reaction is complete by TLC, directly filter the reaction solution, wash with water (20 mL × 3), dry the filter cake by suction and then dry it under vacuum to obtain the target compound 43b (6-bromophthalazine-1,4-diol) (1.0 g, yield: 94.34%, LCMS m / z = 241.1 [M+1] + )。

[0354] Step 2: Dissolve compound (43b) (0.70 g, 2.9 mmol) in POCl3 (4 mL), heat to 110 °C and react overnight. Determine that the reaction is complete by TLC. Concentrate the reaction solution to one-tenth of its volume and then slowly add it dropwise to ice water (20 mL). Stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound 43c (6-bromo-1,4-dichlorophthalazine) (0.61 g, yield: 75.68%, LCMS m / z = 277.0 [M+1] + )。

[0355] Step 3: Dissolve compound (43c) (0.60 g, 2.16 mmol), (R)-1-methylpiperidin-3-amine (0.25 g, 2.16 mmol), and Na2CO3 (0.43 g, 4.00 mmol) in dry DMF (4 mL). Place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound (43d) ((R)-7-bromo-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine) (0.50 g, yield: 65.09%, LCMS m / z = 355.1 [M+1] + )。

[0356] Step 4: At room temperature, add compound (43d) (0.40 g, 1.12 mmol), dimethylphosphine oxide (87.4 mg, 1.12 mmol), palladium acetate (25 mg, 0.11 mmol), Xantphos (92 mg, 0.16 mmol), and anhydrous potassium phosphate (467 mg, 2.2 mmol) to dry DMF (6 mL). Replace with argon 3 times, heat to 150 °C and react for 3 hours. After rotary evaporation of the reaction solution, directly separate by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound 43e ((R)-(1-chloro-4-((1-methylpiperazin-3-yl)amino)phthalazin-6-yl)dimethylphosphine oxide) (32 mg, yield: 7.7%, LCMS m / z = 353.2 [M+1] +) and compound 43f ((R)-4-chloro-1-((1-methylpiperazin-3-yl)amino)phthalazin-6-yl)dimethylphosphine oxide) (30 mg, yield: 7.6%, LCMS m / z = 353.2 [M+1] + ).

[0357] Step Five: Add compound (43e) (30 mg, 0.08 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35 mg, 0.17 mmol), sodium carbonate (26 mg g, 0.24 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1), displace with nitrogen 3 times, and heat to 110 °C for reaction for 16 hours. After rotary evaporation of the reaction solution, directly separate by silica gel column chromatography (DCM:CH3OH = 10:1) to obtain compound 43 ((R)-(1-(2-hydroxy-4-(trifluoromethyl)phenyl)-4-((1-methylpiperidin-3-yl)amino)phthalazin-6-yl)dimethylphosphine oxide) (0.01 g, yield: 24.59%, LCMS m / z = 479.6 [M+1] + ).

[0358] Example 44: Synthesis of (R)-5-fluoro-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol

[0359]

[0360] Step: Add compound (1d) (0.05 g, 0.18 mmol), (4-fluoro-2-hydroxyphenyl)boronic acid (41.85 mg, 0.27 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1), displace with nitrogen 3 times, and heat to 110 °C for reaction for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 44 (R)-5-fluoro-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol (0.02 g, yield: 31.54%, LCMS m / z = 353.2 [M+1] + ).

[0361] Example 45: (R)-(4-(2-Hydroxy-4-(trifluoromethyl)phenyl)-1-((1-methylpiperidin-3-yl)amino)phthalazin-6-yl)dimethylphosphine oxide

[0362]

[0363] Step 1: Add compound (43f) (30 mg, 0.08 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35 mg, 0.17 mmol), sodium carbonate (26 mg, 0.24 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the air with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 45 (R)-(4-(2-Hydroxy-4-(trifluoromethyl)phenyl)-1-((1-methylpiperidin-3-yl)amino)phthalazin-6-yl)dimethylphosphine oxide (0.01 g, yield: 24.59%, LCMS m / z = 479.6 [M+1] + )

[0364] Example 46: Synthesis of (R)-4-Fluoro-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol

[0365]

[0366] Step: Add compound (1d) (0.05 g, 0.18 mmol), (5-fluoro-2-hydroxyphenyl)boronic acid (41.85 mg, 0.27 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the air with nitrogen three times, heat to 110 °C and react for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and perform column chromatography separation (DCM:CH3OH = 10:1) of the residue to obtain the target compound 46 (R)-4-Fluoro-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol (0.012 g, yield: 18.93%, LCMS m / z = 353.2 [M+1] + )

[0367] Example 47: Synthesis of ((R)-4-Methyl-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol

[0368]

[0369] Procedure: Add compound (1d) (0.05 g, 0.18 mmol), (2-Hydroxy-5-methylphenyl)boronic acid (40.77 mg, 0.27 mmol), sodium carbonate (42.40 mg, 0.40 mmol) and Pd(dppf)Cl2 (19.75 mg, 0.027 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the air with nitrogen three times, heat to 110 °C and react for 3 hours. Quench the reaction mixture with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 47 (R)-4-Methyl-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol (0.015 g, yield: 23.%, LCMS m / z = 349.2 [M+1] + )。

[0370] Example 48: 2-(3-Methyl-8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol and

[0371] Example 49: 2-(3-Methyl-5-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol

[0372]

[0373] Step 1: Dissolve compound (48a) (3-Methyl-5H,7H-furo[3,4-b]pyridine-5,7-dione) (1.0 g, 6.13 mmol) and N2H4·H2O (2.5 g, 41.1 mmol, 80% wt) in acetic acid (20 mL), heat to 110 °C and reflux overnight. Determine that the reaction is complete by TLC, directly filter the reaction mixture, wash with water (20 mL × 3), dry the filter cake under vacuum after suction drying, to obtain the target compound 43b (3-Methylpyrido[2,3-d]pyridazine-5,8-diol) (1.0 g, yield: 92.08%, LCMS m / z = 177.1 [M+1] + )。

[0374] Step 2: Dissolve compound (48b) (1.0 g, 5.6 mmol) in POCl3 (5 mL), heat to 110 °C and react overnight. Determine that the reaction is complete by TLC. Concentrate the reaction solution to one-tenth of its volume and then slowly add it dropwise to ice water (20 mL), stir well, adjust the pH to 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound 48c (5,8-dichloro-3-methylpyrido[2,3-d]pyridazine) (0.70 g, yield: 58.66%, LCMS m / z = 214.2 [M+1] + )

[0375] Step 3: Dissolve compound (48c) (0.46 g, 2.16 mmol), (R)-1-methylpiperidin-3-amine (0.25 g, 2.16 mmol), and Na2CO3 (0.43 g, 4.00 mmol) in dry DMF (4 mL). Place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC. Add the reaction solution to water (20 mL), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 48d ((R)-5-chloro-3-methyl-N-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyridazin-8-amine) (0.20 g, yield: 31.73%, LCMS m / z = 292.2 [M+1] + ) and compound 48e ((3R)-N-(8-chloro-3-methylpyrido[2,3-d]pyridazin-5-yl)-1-methylpiperidin-3-amine) (50 mg, yield: 7.95%, LCMS m / z = 292.2 [M+1] + )

[0376] Step 4: Add compound (48d) (200 mg, 0.69 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (280 mg, 1.38 mmol), sodium carbonate (220 mg, 2.07 mmol) and Pd(dppf)Cl2 (50 mg, 0.07 mmol) into a mixed solvent of 10 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 48 (2-(3-methyl-8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol) (0.10 g, yield: 34.72%, LCMS m / z = 418.6 [M+1] + ).

[0377] Step 5: Add compound (48e) (25 mg, 0.086 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35 mg, 0.17 mmol), sodium carbonate (28 mg, 0.26 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into a mixed solvent of 5 mL dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 49 (2-(3-methyl-5-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol) (20 mg, yield: 55.71%, LCMS m / z = 418.6 [M+1] + ).

[0378] Example 50: (R)-2-(3-ethyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol and

[0379] Example 51: (R)-2-(3-ethyl-5-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol

[0380]

[0381] Step 1: Dissolve compound (50a) 3-ethylfuro[3,4-b]pyridine-5,7-dione (1.0 g, 5.64 mmol), N2H4·H2O (2.5 g, 41.1 mmol, 80% wt) in acetic acid (20 mL), heat to 110 °C and reflux overnight. Determine that the reaction is complete by TLC, directly filter the reaction solution, wash with water (20 mL × 3), dry the filter cake by suction and then dry it under vacuum to obtain the target compound 50b (3-ethylpyrido[2,3-d]pyridazine-5,8-diol) (0.8 g, yield: 74.26%, LCMS m / z = 192.2 [M+1] + ).

[0382] Step 2: Dissolve compound (50b) (0.8 g, 4.2 mmol) in POCl3 (5 mL), heat to 110 °C and react overnight. Determine that the reaction is complete by TLC, concentrate the volume of the reaction solution to one-tenth and then slowly add it dropwise to ice water (20 ml), stir well, adjust the pH = 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound 50c (5,8-dichloro-3-ethylpyrido[2,3-d]pyridazine) (0.70 g, yield: 73.09%, LCMS m / z = 228.1 [M+1] + ).

[0383] Step 3: Dissolve compound (50c) (0.46 g, 2.01 mmol), (R)-1-methylpiperidin-3-amine (0.25 g, 2.16 mmol), Na2CO3 (0.43 g, 4.00 mmol) in dry DMF (4 mL), place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC, add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 50d ((R)-5-chloro-3-ethyl-N-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyridazin-8-amine) (0.20 g, yield: 32.51%, LCMS m / z = 306.2 [M+1] + ) and compound 50e (R)-8-chloro-3-ethyl-N-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyridazin-5-amine (100 mg, yield: 16.25%, LCMS m / z = 305.4 [M+1] + ).

[0384] Step 4: Add compound (50d) (100 mg, 0.326 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (280 mg, 1.38 mmol), sodium carbonate (220 mg, 2.07 mmol) and Pd(dppf)Cl2 (50 mg, 0.07 mmol) to a mixed solvent of 10 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 50 ((R)-2-(3-ethyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol) (0.05 g, yield: 35.56%, LCMS m / z = 432.6 [M+1] + )

[0385] Step 5: Add compound (50e) (100 mg, 0.326 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35 mg, 0.17 mmol), sodium carbonate (28 mg, 0.26 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 51 ((R)-2-(3-ethyl-5-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol) (0.1 g, yield: 71.17%, LCMS m / z = 432.6 [M+1] + )

[0386] Example 52: (R)-2-(2-methyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol and

[0387] Example 53: (R)-2-(2-methyl-5-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol

[0388]

[0389] Step 1: Dissolve compound (52a) (2-methylfuro[3,4-b]pyridine-5,7-dione) (1.0 g, 6.13 mmol) and N2H4·H2O (2.5 g, 41.1 mmol, 80% wt) in acetic acid (20 mL), heat to 110 °C and reflux overnight. Determine that the reaction is complete by TLC, directly filter the reaction solution, wash with water (20 mL × 3), dry the filter cake by suction and then dry it under vacuum to obtain the target compound 52b (2-methylpyrido[2,3-d]pyridazine-5,8-diol) (1.0 g, yield: 92.08%, LCMS m / z = 177.1 [M+1] + )

[0390] Step 2: Dissolve compound (52b) (1.0 g, 5.6 mmol) in POCl3 (5 mL), heat to 110 °C and react overnight. Determine that the reaction is complete by TLC, concentrate the volume of the reaction solution to one-tenth and then slowly add it dropwise to ice water (20 ml), stir well, adjust the pH = 8 with 2N sodium hydroxide aqueous solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound 52c (5,8-dichloro-2-methylpyrido[2,3-d]pyridazine) (0.70 g, yield: 58.66%, LCMSm / z = 214.2 [M+1] + )

[0391] Step 3: Dissolve compound (52c) (0.46 g, 2.16 mmol), (R)-1-methylpiperidin-3-amine (0.25 g, 2.16 mmol), and Na2CO3 (0.43 g, 4.00 mmol) in dry DMF (4 mL), place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC, add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 52d ((R)-5-chloro-2-methyl-N-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyridazin-8-amine) (0.10 g, yield: 15.90%, LCMS m / z = 292.4 [M+1] + ) and compound 52e ((R)-8-chloro-2-methyl-N-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyridazin-5-amine) (0.10 g, yield: 15.90%, LCMS m / z = 292.4 [M+1] + )

[0392] Step 4: Add compound (52d) (100 mg, 0.34 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (280 mg, 1.38 mmol), sodium carbonate (220 mg, 2.07 mmol) and Pd(dppf)Cl2 (50 mg, 0.07 mmol) to a mixed solvent of 10 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After evaporating the reaction solution to dryness, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 52 ((R)-2-(2-methyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethyl)phenol) (25 mg, yield: 17.63%, LCMS m / z = 418.6 [M+1] + )

[0393] Step 5: Add compound (52e) (100 mg, 0.34 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (35 mg, 0.17 mmol), sodium carbonate (28 mg, 0.26 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After evaporating the reaction solution to dryness, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 53 ((R)-2-(2-methyl-5-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-8-yl)-5-(trifluoromethyl)phenol) (20 mg, yield: 14.10%, LCMS m / z = 418.6 [M+1] + )

[0394] Example 54: (R)-2-(4-((1-methylpiperidin-3-yl)amino)thieno[3,4-d]pyridazin-1-yl)-5-(trifluoromethyl)phenol

[0395]

[0396] Step 1: Dissolve compound (54a) (3,4-thiophenedicarboxylic anhydride) (180 mg, 1.16 mmol), N2H4·H2O (0.6 mL, 9.88 mmol, 80% wt) in acetic acid (10 mL), heat to 110 °C and reflux overnight. Determine that the reaction is complete by TLC, directly filter the reaction solution, wash with water (10 mL × 3), dry the filter cake by suction and then dry it under vacuum to obtain the target compound 54b (thieno[3,4-d]pyridazine-1,4-diol) (150 mg, yield: 76.89%, LCMS m / z = 169.1 [M+1] + )。

[0397] Step 2: Dissolve compound (54b) (150 mg, 0.89 mmol) in POCl3 (3 mL), heat to 110 °C and react overnight. Determine that the reaction is complete by TLC, concentrate the volume of the reaction solution to one-tenth and then slowly add it dropwise to ice water (10 ml), stir well, adjust the pH = 8 with 2N aqueous sodium hydroxide solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (PE:EA = 20:1) to obtain the target compound 54c (1,4-dichlorothieno[3,4-d]pyridazine) (76 mg, yield: 41.64%, LCMS m / z = 205.0 [M+1] + )。

[0398] Step 3: Dissolve compound (54c) (70 mg, 0.34 mmol), (R)-1-methylpiperidin-3-amine (40 mg, 0.35 mmol), Na2CO3 (74 mg, 0.70 mmol) in dry DMF (3 mL), place this mixed system in a sealed tube and heat to 120 °C and react overnight. Determine that the raw materials are completely converted by TLC, add the reaction solution to water (10 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compound 54d ((R)-4-chloro-N-(1-methylpyridin-3-yl)thieno[3,4-d]pyridazin-1-amine) (50 mg, yield: 52.00%, LCMS m / z = 283.1 [M+1] + )。

[0399] Step 4: Add compound (54d) (50 mg, 0.18 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (74 mg, 0.36 mmol), sodium carbonate (60 mg, 0.57 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) to 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 54 ((R)-2-(4-((1-methylpiperidin-3-yl)amino)thieno[3,4-d]pyridazin-1-yl)-5-(trifluoromethyl)phenol) (20 mg, yield: 27.2%, LCMS m / z = 409.4 [M+1] + ).

[0400] Example 55: (R)-5-Fluoro-2-(3-methyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)phenol

[0401]

[0402] Step 1: At room temperature, add compound (48d) (30 mg, 0.10 mmol), 5-fluoro-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (86 mg, 0.36 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) to 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 55 ((R)-5-fluoro-2-(3-methyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)phenol) (20 mg, yield: 45.36%, LCMS m / z = 368.6 [M+1] + )

[0403] Example 56: (R)-2-(3-methyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)phenol

[0404]

[0405] Step 1: At room temperature, add compound (48d) (30 mg, 0.10 mmol), 5-fluoro-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (79 mg, 0.36 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 56 ((R)-2-(3-methyl-8-((1-methylpiperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)phenol) (30 mg, yield: 71.55%, LCMS m / z = 350.6 [M+1] + )

[0406] Example 60: 5-Methyl-2-(3-methyl-8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)phenol

[0407]

[0408] Step 1: At room temperature, add compound (48d) (30 mg, 0.10 mmol), 5-methyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (47 mg, 0.20 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) into 5 mL of a mixed solvent of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, heat to 110 °C and react for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 60 (5-methyl-2-(3-methyl-8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)phenol) (10 mg, yield: 27.51%, LCMS m / z = 364.6 [M+1] + )

[0409] Example 61: 2-(3-Methyl-8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethoxy)phenol

[0410]

[0411] Step 1: At room temperature, add compound (48d) (30 mg, 0.10 mmol), 5-trifluoromethoxy-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (61 mg, 0.20 mmol), sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2 (5 mg, 0.007 mmol) to a mixed solvent of 5 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times, and heat to 110 °C for reaction for 16 hours. After rotary evaporation of the reaction solution, directly perform silica gel column chromatography separation (DCM:CH3OH = 10:1) to obtain compound 61 (2-(3-methyl-8-{[(3R)-1-methylpiperidin-3-yl]amino}pyrido[2,3-d]pyridazin-5-yl)-5-(trifluoromethoxy)phenol) (20 mg, yield: 46.14%, LCMS m / z = 434.6 [M+1] + )

[0412] Example 62: ((R)-2-(1-Methyl-7-((1-methylpiperidin-3-yl)amino)-1H-pyrazolo[3,4-d]pyridazin-4-yl)-5-(trifluoromethyl)phenol) and

[0413] Example 63: ((R)-2-(1-Methyl-4-((1-methylpiperidin-3-yl)amino)-1H-pyrazolo[3,4-d]pyridazin-7-yl)-5-(trifluoromethyl)phenol)

[0414]

[0415] Step 1: Under an ice-water bath, dissolve compound (62a) (10.0 g, 69.84 mmol), DMAP (0.14 g, 1.25 mmol), and triethylamine (9.19 g, 90.79 mmol) in dry tetrahydrofuran (100 mL). Slowly add a tetrahydrofuran solution (50 mL) of ethyl 2-chloro-2-oxoacetate (11.44 g, 83.81 mmol) dropwise to this mixture. After addition, allow it to naturally rise to room temperature and react for 1 hour. TLC shows that the raw material (62a) is consumed. Filter the salt formed in the reaction solution, add the filtrate back to the reaction flask, add tert-butyl 1-methylhydrazine-1-carboxylate (12.25 g, 83.81 mmol), and continue to react at room temperature for 2 hours after addition. Directly concentrate the reaction solution to dryness to obtain a yellow solid. This solid is recrystallized with methanol / water (100 mL, v / v = 1:1) and then dried under vacuum to obtain the target compound 62b ((Z / E mixture)-diethyl 2-(2-(tert-butoxycarbonyl)-2-methylhydrazino)methylene)-3-oxosuccinate) (19.2 g, yield: 79.83%)

[0416] Step 2: The raw material (62b) (19.2 g, 55.75 mmol) was dispersed in ethyl acetate (60 ml), and HCl / dioxane solution (200 ml) was added. This mixed solution was heated to 50 °C and reacted for 3 hours. It was concentrated to dryness, redissolved in ethyl acetate (200 ml), adjusted to pH > 7 with 2N aqueous sodium carbonate solution, separated by liquid-liquid extraction, the organic phase was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, the solvent in the filtrate was removed under reduced pressure, and the residue was separated by column chromatography (PE:EA = 5:1) to obtain the target compound 62c (diethyl 1-methyl-1H-pyrazole-4,5-dicarboxylate) (10.2 g, yield: 80.87%, LCMS m / z = 227.2 [M+1]+)

[0417] Step 3: Dimethyl 1-methyl-1H-pyrazole-4,5-dicarboxylate (62c) (0.8 g, 4.04 mmol) and N2H4.H2O (0.61 g, 12.12 mmol) were dissolved in acetic acid (5 mL), and the mixture was heated to 120 °C and refluxed overnight. TLC confirmed the completion of the reaction. The reaction solution was directly concentrated to dryness, then dispersed in water (30 ml) to form a solid, filtered, the filter cake was dried by suction and then dried under vacuum overnight to obtain the target compound 62d (1-methyl-1H-pyrazolo[3,4-d]pyridazine-4,7-diol) (0.40 g, yield: 59.59%, LCMS m / z = 167.2 [M+1] + )。

[0418] Step 4: Compound 62d (0.40 g, 2.41 mmol) was dissolved in POCl3 (10 mL), and the mixture was heated to 100 °C and reacted overnight. TLC confirmed the completion of the reaction. The reaction solution was directly concentrated, and after removing POCl3, the crude oil was slowly added dropwise to ice water (50 ml), stirred well, adjusted to pH = 8 with 2N aqueous sodium hydroxide solution, extracted with ethyl acetate (30 mL × 3), the combined organic phases were washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, the solvent in the filtrate was removed under reduced pressure, and the residue was separated by column chromatography (PE:EA = 5:1) to obtain the target compound (62e) (4,7-dichloro-1-methyl-1H-pyrazolo[3,4-d]pyridazine) (0.40 g, yield: 81.75%, LCMS m / z = 203.2 [M+1] + )。

[0419] Step 5: Dissolve compound (62e) (0.40 g, 1.97 mmol), (R)-1-methylpiperidin-3-amine (0.27 g, 2.36 mmol), and Na2CO3 (0.21 g, 1.97 mmol) in dry DMF (5 mL). Place this mixed system in a sealed tube and heat it to 120 °C for overnight reaction. TLC was used to determine that the raw materials were completely converted. Add the reaction solution to water (20 ml), stir well, extract with ethyl acetate (10 mL × 3), combine the organic phases, wash with saturated brine (10 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, and separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain the target compounds (a mixture of 62f and 63f) (((R)-4-chloro-1-methyl-N-(1-methylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyridazin-7-amine, (R)-7-chloro-1-methyl-N-(1-methylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyridazin-4-amine) (0.31 g, yield: 56.04%, LCMS m / z = 281.2 [M+1] + ).

[0420] Step 6: Add compound (a mixture of 62f and 63f) (0.31 g, 1.10 mmol), (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (350.00 mg, 1.72 mmol), sodium carbonate (220.10 mg, 2.22 mmol), and Pd(dppf)Cl2 (90.75 mg, 0.13 mmol) to a mixed solvent of 30 mL of dioxane and water (v / v = 4:1). Replace the gas with nitrogen three times and heat to 110 °C for 3 hours. Quench the reaction solution with water (50 mL), extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure from the filtrate, separate the residue by column chromatography (DCM:CH3OH = 10:1) to obtain a mixture of target compounds (a mixture of 62 and 63), and then separate by preparative pre-HPLC to obtain target compound 62 ((R)-2-(1-methyl-7-((1-methylpiperidin-3-yl)amino)-1H-pyrazolo[3,4-d]pyridazin-4-yl)-5-(trifluoromethyl)phenol) (0.04 g, yield: 8.91%, LCMS m / z = 407.4 [M+1] + ), target compound 63 ((R)-2-(1-methyl-4-((1-methylpiperidin-3-yl)amino)-1H-pyrazolo[3,4-d]pyridazin-7-yl)-5-(trifluoromethyl)phenol) (0.16 g, yield: 35.66%, LCMS m / z = 407.4 [M+1] + ).

[0421] Example 26 and 28 are similar to Example 25, replacing the amine in Step 1 with the corresponding commercially available starting material amine.

[0422] Examples 57 - 59 are similar to Example 36, replacing the imidazole dicarboxylate in Step 2 with the corresponding commercially available starting material, and replacing it with the required boric acid or borate ester in Step 5.

[0423] Examples 64 - 75 are similar to Example 62, replacing the amine in Step 5 with the required commercially available amine, and replacing it with the required boric acid or borate ester in Step 6.

[0424] Examples 76 - 88 are similar to Example 36, replacing the amine in Step 4 with the required commercially available amine, and replacing it with the required boric acid or borate ester in Step 5.

[0425] Table 1. Data Characterization of Example Compounds

[0426]

[0427]

[0428]

[0429]

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436]

[0437]

[0438]

[0439] Example 89: Pyroptosis Test

[0440] The in vitro activities of the above compounds were demonstrated in the following assays:

[0441] In vitro activity screening of small molecule compounds targeting the inhibition of NLRP3 inflammasome was carried out using THP-1 human monocytic cells (THP-1). THP-1 can be induced to differentiate into macrophages by phorbol 12-myristate 13-acetate (PMA), and then macrophage M1 polarization can be induced by lipopolysaccharide (LPS) to release cytokines such as TNF-α and IL-6, thus establishing a typical inflammation model.

[0442] 1. Experimental materials:

[0443] RPMI Medium 1640 culture medium was purchased from Gibco, penicillin and streptomycin were purchased from Hyclone, lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA), and nigericin were purchased from MedChemExpress (MCE), thiazolyl blue (MTT) was purchased from Beijing Solarbio Science & Technology Co., Ltd., and sodium dodecyl sulfate (SDS) was purchased from Biofroxx.

[0444] 2. THP-1 cell culture:

[0445] THP-1 cells were cultured in 1640 medium + 10% FBS + 1% penicillin / streptomycin medium in an incubator at 37°C and 5% CO2.

[0446] 3. THP-1 cell pyroptosis test:

[0447] THP-1 cells in the logarithmic growth phase were collected to prepare a cell suspension of 1×10 6 cells / mL. At the same time, PMA was added to make its final concentration in the cell suspension 300 ng / mL, and then inoculated into a 96-well plate at 1×10 5 cells per well and cultured in a cell incubator at 37°C and 5% CO2 for 24 hours to induce the cells to differentiate into macrophages.

[0448] On the next day, LPS was added to stimulate the cells to induce an inflammatory model. The specific operation was as follows: Remove the original culture medium in the well plate, add 100 μL of 1640 medium containing 2 μg / mL LPS to each well, and then place the 96-well plate in a cell culture incubator at 37°C and 5% CO2 for 3 - 4 hours to establish an inflammatory model. In each 96-well plate, a drug treatment group, an inflammatory model group, a normal cell group (containing only cells and 1640 medium), and a blank control group (containing only medium without cells) were set up. After the LPS stimulation ended, the test compound was diluted to the corresponding concentration (0.004 - 40 μM) with 1640 medium and added to the corresponding wells of the 96-well plate, 50 μL per well, with 3 replicates for each sample concentration; the inflammatory model group and the normal cell group were added with 50 μL of 1640 medium, and then the 96-well plate was placed in a cell culture incubator at 37°C and 5% CO2 for 30 min. After the culture ended, 50 μL of 1640 medium containing 40 μM nigericin was added to the drug treatment group and the inflammatory model group respectively. Nigericin can activate the NLRP3 inflammasome of the cells and induce pyroptosis; the normal cell group was added with 50 μL of 1640 medium. At this time, in the 200 μL system in the 96-well plate, the final concentration of each drug was 0.001 - 10 μM, and the final concentration of nigericin was 10 μM. After adding nigericin, the 96-well plate was placed in a cell culture incubator at 37°C and 5% CO2 for 3 - 4 hours. After the culture ended, 20 μL of MTT solution (5 mg / mL) was added to each well, and it was incubated in a cell culture incubator at 37°C and 5% CO2 for 1.5 h. Then, 50 μL of 20% SDS solution (containing 0.1% hydrochloric acid) was added to each well, and the 96-well plate was placed in a cell culture incubator at 37°C and 5% CO2 and incubated overnight. On the third day, the absorbance was measured at a wavelength of 562 nm using an enzyme-linked immunosorbent assay (ELISA) reader. And the pyroptosis protection rate of the drug on the cells was calculated according to the following formula:

[0449] Pyroptosis protection rate of cells = [(X - C0) / (C - C0)] × 100%

[0450] Among them, C, C0, and X represent the average absorbance values of the normal cell group, the blank control group, and the drug treatment group, respectively. Finally, the Graphpad Prism 5.0 software was used to fit the cell survival rate curve and calculate the EC50 value of the test compound inhibiting pyroptosis of cells caused by the NLRP3 inflammasome.

[0451] Table 2: EC50 values of the compounds in each example in vitro determination 1

[0452]

[0453]

[0454] For the EC50 value, "+" indicates that the EC50 value is greater than 1 μM, "++" indicates that the EC50 value is greater than 500 nM and less than or equal to 1 μM, "+++" indicates that the EC50 value is greater than 100 nM and less than or equal to 500 nM; "++++" indicates that the EC50 value is greater than 20 nM and less than or equal to 100 nM; "+++++" indicates that the EC50 value is less than 20 nM.

Claims

1. A compound represented by the following chemical formula (IVd) or a pharmaceutically acceptable salt thereof: Wherein: R3 is selected from hydrogen, methyl, cyclopropyl, ethyl, fluorine, trifluoromethyl; A is a single bond, -CH2-, -(CH3)CH-, -CH2CH2-; R4 is selected from R 11 selected from trifluoromethyl, methyl, trifluoromethoxy, fluorine; Among them, the following compounds are not included:

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R3 is selected from hydrogen, methyl, cyclopropyl, trifluoromethyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, A is a single bond, -CH2-, -CH2CH2-.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 11 Selected from trifluoromethyl, fluorine.

5. The following compound or a pharmaceutically acceptable salt thereof, 6. A pharmaceutical composition, characterized in that, The composition comprises a compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

7. Use of a compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition of claim 6, in the preparation of a medicament for treating NLRP3-mediated diseases.

Images