P2x7 receptor antagonist

Abstract

The present application relates to the field of chemical drugs, and discloses a P2X7 receptor antagonist. The structure of the P2X7 receptor antagonist is as shown in formula I. The present application has found that the compound shown in formula I not only has excellent antagonistic activity for the P2X7 receptor, but also has excellent metabolic stability and pharmacokinetic properties. Compared with the positive control JNJ47965567, the metabolic stability and pharmacokinetic properties of the compound of the present application are significantly improved, and the druggability is significantly better. The compound shown in formula I is used as a P2X7 receptor antagonist, and can be used to prepare a drug for treating illnesses such as inflammation and inflammation-related diseases, kidney diseases, respiratory diseases, cancer, pain, central nervous system diseases, radiation-induced brain injury, cerebral ischemia, myocardial injury, diabetes, depression, lupus erythematosus, atherosclerosis, and allergic asthma, and has broad application prospects.

Description

P2X7 receptor antagonists Technical Field

[0001] The present application belongs to the field of chemical drugs, and specifically relates to P2X7 receptor antagonists. Background Art

[0002] Buenstock et al. discovered that ATP exerts its effects through its action on the purinergic receptor P2, and classified ATP-sensitive P2 receptors into two types: ligand-gated ion channels (P2X) and G-protein-coupled P2Y. Based on differences in P2X receptor components and signal transduction pathways, seven subtypes (P2X1-7) have been cloned in mammalian cells.

[0003] P2X7 is a specialized subtype of the purinergic receptor P2X, a nonselective cation channel receptor activated by high concentrations of extracellular ATP. P2X receptors share a basic structure, consisting of two transmembrane domains: an intracellular amino-terminus (N-terminus) and a carboxyl-terminus (C-terminus), and an extracellular loop of cysteine ​​residues. However, the P2X7 receptor comprises 595 amino acid residues, with its C-terminus being the longest of all family members (consisting of 239 amino acids). This unique molecular structure not only enables it to function as an ion channel but also, upon activation, to transform from an ion channel into a larger cellular pore, participating in physiological and pathological processes such as transmitter release, signal transduction, and inflammatory responses. P2X7 receptors are widely distributed in mammals. Upon activation, they induce the maturation and release of cytokines such as IL-1β and IL-18, plasma membrane reorganization, extracellular domain shedding, and cell death, potentially acting as a "crisis response sensor" in the body. The P2X7 receptor is widely distributed in tissues and organs such as the brain, kidney, liver, lung, and spleen, and its expression can be seen in a variety of immune-related cells such as monocytes, macrophages, lymphocytes, mast cells, and neutrophils. Due to its wide distribution, the P2X7 receptor is associated with a variety of diseases such as central nervous system (CNS) diseases, inflammatory bowel disease, hepatitis, inflammatory pain, kidney injury, rheumatoid arthritis (RA), and cardiovascular disease, and is an emerging target for the treatment of inflammation and other related diseases.

[0004] JNJ47965567 is a commercially available P2X7 receptor antagonist with good antagonistic effects on both human and rat P2X7 receptors. However, JNJ47965567 has poor metabolic stability and pharmacokinetic properties, which reduce its therapeutic efficacy.

[0005] In order to overcome the above problems, a compound that can not only effectively antagonize the P2X7 receptor but also has excellent metabolic stability and pharmacokinetic properties has been developed, which is of great significance for the treatment of various diseases including inflammation-related diseases, kidney-related diseases, and central nervous system diseases. Summary of the Invention

[0006] The purpose of this application is to provide a new P2X7 receptor antagonist.

[0007] The present application provides a compound represented by Formula I, a salt thereof, an isotope compound thereof, a stereoisomer thereof, or a hydrate thereof:

[0008] in,

[0009] n is 0, 1, 2, or 3;

[0010] R1 is selected from one or more R 1a The following groups substituted: 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 8-membered saturated cycloalkyl, 3- to 8-membered saturated heterocyclic group; R 1a Each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, nitro, carboxyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, SO2R 1b ; R 1b Selected from C 1～8 alkyl;

[0011] Y is CH;

[0012] X is N, R2 is nothing;

[0013] R3 is selected from hydrogen, cyano, halogen, hydroxy, amino, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 10-membered aryl or 5- to 10-membered heteroaryl;

[0014] R4 is selected from the following groups substituted by one or more substituents: 5-6 membered aryl, 5-6 membered heteroaryl, fused heterocyclic group; the substituents are each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, SH, j R h Substituted with the following groups: SR k , 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 5- to 6-membered saturated cycloalkyl, 5- to 6-membered saturated heterocyclic group, R k Selected from 5-6 membered aryl, 5-6 membered heteroaryl,

[0015] j is 0, 1, 2, or 3;

[0016] R h Each independently selected from hydrogen, halogen, hydroxyl, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, cyano, amino, NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl, 5-6 membered heteroaryl, R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

[0017] In one embodiment, R1 is selected from one or more R 1a Substituted groups include 5- to 6-membered aryl and 5- to 6-membered heteroaryl.

[0018] In one embodiment, R1 is selected from one or more R 1a Substituted with the following groups:

[0019] In one embodiment, R1 is selected from one or more R 1a Substituted with the following groups: R 1a are each independently selected from halogen, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, 3- to 8-membered cycloalkyl.

[0020] In one embodiment, R 1a Each is independently selected from halogen, trifluoromethyl, methoxy, and cyclopropyl.

[0021] In one embodiment, the halogen is selected from chlorine and bromine.

[0022] In one embodiment, R3 is selected from hydrogen, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy.

[0023] In one embodiment, R3 is selected from hydrogen, C 1～8alkyl.

[0024] In one embodiment, R3 is selected from C 1～8 alkyl.

[0025] In one embodiment, R3 is selected from methyl.

[0026] In one embodiment, R4 is selected from a 5- to 6-membered heteroaryl group or a fused heterocyclic group substituted with one or two or more of the above substituents.

[0027] In one embodiment, R4 is selected from the following groups substituted with one or two or more substituents:

[0028] In one embodiment, R4 is selected from a 5-membered heteroaryl group substituted with one or two or more of the substituents.

[0029] In one embodiment, R4 is selected from the following groups substituted with one or two or more substituents:

[0030] In one embodiment, the substituent is selected from j R h Substituted groups include: 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 5- to 6-membered saturated cycloalkyl, and 5- to 6-membered saturated heterocyclic group.

[0031] In one embodiment, the substituent is selected from j R h Substituted 6-membered aryl.

[0032] In one embodiment, j is 1, and the R h Selected from NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl, 5-6 membered heteroaryl; R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

[0033] In one embodiment, the R h Selected from MSO2R 4c , M is selected from 5- to 6-membered aryl and 5- to 6-membered heteroaryl, R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

[0034] In one embodiment, the structure of the compound is as shown in Formula II:

[0035] Where n is 0, 1 or 2;

[0036] Y6 is selected from N, CR s6 ; Y7 is selected from N, CR s7 ; Y8 is selected from N, CR s8 ; Y9 is selected from N, CR s9 ; Y 10 Selected from N, CR s10 ;

[0037] R s6 、R s7 、R s8 、R s9 、R s10 are independently selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, carboxyl, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy, 3- to 8-membered cycloalkyl;

[0038] R3 is selected from hydrogen, cyano, halogen, hydroxy, amino, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 10-membered aryl or 5- to 10-membered heteroaryl;

[0039] R4 is selected from the following groups substituted by one or more substituents: 5-6 membered aryl, 5-6 membered heteroaryl, fused heterocyclic group; the substituents are each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, SH, j R h Substituted with the following groups: SR k , 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 5- to 6-membered saturated cycloalkyl, 5- to 6-membered saturated heterocyclic group, R k Selected from 5-6 membered aryl, 5-6 membered heteroaryl,

[0040] j is 0, 1, 2, or 3;

[0041] R h Each independently selected from hydrogen, halogen, hydroxyl, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C1～8 Alkoxy, cyano, amino, NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl, 5-6 membered heteroaryl, R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

[0042] In one embodiment, Y6 is N; Y7 is CR s7 ; Y8 is CR s8 ; Y9 is CR s9 ; Y 10 CR s10 ; R s6 、R s7 、R s8 、R s10 are each independently selected from hydrogen; R s9 Selected from halogen, hydroxy, amino, cyano, nitro, carboxyl, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy, 3-8 membered cycloalkyl; R s9 Preferred are halogen, trifluoromethyl, methoxy, and cyclopropyl.

[0043] In one embodiment, Y7 is N; Y6 is CR s6 ; Y8 is CR s8 ; Y9 is CR s9 ; Y 10 CR s10 ; R s6 、R s7 、R s8 、R s9 are each independently selected from hydrogen; R s10 Selected from halogen, hydroxy, amino, cyano, nitro, carboxyl, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy, 3-8 membered cycloalkyl; R s10 Preferred are halogen, trifluoromethyl, methoxy, and cyclopropyl.

[0044] In one embodiment, the structure of the compound is as shown in Formula II-1, Formula II-2, or Formula II-3:

[0045] Where n is 0 or 1;

[0046] Y6 is selected from N, CR s6 ; Y7 is selected from N, CR s7 ; Y8 is selected from N, CR s8 ; Y9 is selected from N, CR s9 ; Y 10 Selected from N, CR s10 ;

[0047] R s6 、R s7 、R s8 、R s9 、R s10 are independently selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, carboxyl, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy, 3- to 8-membered cycloalkyl;

[0048] R3 is selected from hydrogen, C 1～5 Alkyl, 3-5 membered cycloalkyl;

[0049] R 4a and R 4b Each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, with j R h Substituted groups: 5-6 membered aryl, 5-6 membered heteroaryl, 5-6 membered saturated cycloalkyl, 5-6 membered saturated heterocyclic group,

[0050] j is 0, 1, 2, or 3;

[0051] R h Each independently selected from hydrogen, halogen, hydroxyl, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, cyano, amino, NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl, 5-6 membered heteroaryl; R4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

[0052] In one embodiment, the substituent is selected from j R h Substituted groups include: 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 5- to 6-membered saturated cycloalkyl, and 5- to 6-membered saturated heterocyclic group.

[0053] In one embodiment, the R 4a and R 4b Each independently selected from j R h Substituted 6-membered aryl.

[0054] In one embodiment, j is 1, and the R h Selected from NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl, 5-6 membered heteroaryl; R 4c Selected from C 1～8 Alkyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclic group; the R h Selected from MSO2R 4c , M is selected from 5- to 6-membered aryl and 5- to 6-membered heteroaryl, R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

[0055] In one embodiment, R 4a Selected from hydrogen, R 4b Selected from hydrogen,

[0056] In one embodiment, the structure of the compound is as shown in Formula II-4 or Formula II-5:

[0057] Wherein, R3 is selected from hydrogen, C 1～3 Alkyl; R S Selected from hydrogen, halogen, C 1～3 Alkyl, halogenated C 1～ 3 alkyl, C 1～3 Alkoxy, halogenated C 1～3 Alkoxy, 3-5 membered saturated cycloalkyl;

[0058] R s Selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8Alkyl, nitro, carboxyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, SO2R 1b ; R 1b Selected from C 1～8 alkyl.

[0059] In one embodiment, the structure of the compound is as shown in Formula II-6, Formula II-7, Formula II-8 or Formula II-9:

[0060] Among them, R s Selected from hydrogen, halogen, C 1～3 Alkyl, halogenated C 1～3 Alkyl, C 1～3 Alkoxy, halogenated C 1～3 Alkoxy, 3- to 5-membered saturated cycloalkyl.

[0061] In one embodiment, R s Selected from halogen, trifluoromethyl, methoxy, cyclopropyl.

[0062] In one embodiment, R s Selected from halogen, trifluoromethyl.

[0063] In one embodiment, the structure of the compound is as shown in Formula II-6-1, Formula II-7-1, Formula II-8-1 or Formula II-9-1:

[0064] Among them, R s is selected from halogen, trifluoromethyl, methoxy, cyclopropyl; in one embodiment, R s Selected from halogen, trifluoromethyl.

[0065] In one embodiment, the compound is one of the following compounds:

[0066] The present application also provides a pharmaceutical composition, which is a preparation prepared by using the above-mentioned compound, its salt, its isotope compound, its stereoisomer or its hydrate as an active ingredient, and adding pharmaceutically acceptable excipients.

[0067] The present application also provides the use of the above-mentioned compound, its salt, its isotope compound, its stereoisomer or its hydrate in the preparation of P2X7 receptor antagonists.

[0068] Preferably, the P2X7 receptor antagonist is a drug that antagonizes the activity of the P2X7 receptor.

[0069] Preferably, the P2X7 receptor antagonist is a drug for treating inflammation and inflammation-related diseases, kidney disease, gout, respiratory diseases, cancer, pain, central nervous system diseases, radiation brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis, and allergic asthma.

[0070] Preferably, the inflammation and inflammation-related diseases are selected from neurological inflammation, arthritis, colitis, pancreatitis fibrosis, alcoholic fatty liver disease, bronchitis, pneumonia, lumbar spondylitis, and vasculitis.

[0071] Preferably, the kidney disease is selected from primary glomerular disease, secondary glomerular disease, interstitial nephritis, tubular disease, renal vascular disease, hereditary kidney disease, acute kidney injury caused by ischemia-reperfusion, acute kidney injury caused by sepsis, drug-induced acute kidney injury, contrast agent-induced acute kidney injury, and chronic renal failure; more preferably, the primary glomerular disease is selected from focal segmental glomerulosclerosis, crescentic nephritis, minimal change disease, IgA nephropathy, and membranous nephropathy; more preferably, the secondary glomerular disease is selected from ANCA-associated vasculitis, hypertensive nephropathy, diabetic nephropathy, hepatitis B-related nephritis, lupus nephritis, purpura nephritis, hyperuricemia, and lipoprotein nephropathy; more preferably, the hereditary kidney disease is selected from polycystic kidney disease; more preferably, the drug-induced acute kidney injury is selected from cisplatin-induced acute kidney injury, folic acid-induced acute kidney injury, and aristolochic acid-induced acute kidney injury.

[0072] Preferably, the gout is selected from acute gouty arthritis, intermittent gout, chronic gout, hyperuricemia, secondary gout, gouty nephropathy, and urinary tract stones.

[0073] Preferably, the respiratory disease is selected from airway obstructive diseases, more preferably bronchial asthma, allergic asthma, intrinsic asthma, exogenous asthma, exercise-induced asthma, and drug-induced asthma.

[0074] Preferably, the cancer is selected from prostate cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, intestinal cancer, colon cancer, gastric cancer, skin cancer, brain tumor, leukemia, lymphoma.

[0075] Preferably, the pain is selected from headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone pain, pain caused by cancer and tumor invasion, and neuropathic pain syndrome.

[0076] Preferably, the central nervous system disease is selected from Alzheimer's disease, Parkinson's syndrome, epilepsy, multiple sclerosis and other demyelinating syndromes, cerebral atherosclerosis, and myasthenia gravis.

[0077] Preferably, the lupus erythematosus is systemic lupus erythematosus.

[0078] The present application also provides the use of the compound, its salt, its isotope compound, its stereoisomer or its hydrate in treating diseases related to P2X7 receptors.

[0079] Preferably, the diseases associated with the P2X7 receptor are drugs for inflammation and inflammation-related diseases, kidney diseases, respiratory diseases, cancer, pain, central nervous system diseases, radiation brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis, and allergic asthma.

[0080] Preferably, the inflammation and inflammation-related diseases are selected from neurological inflammation, arthritis, colitis, pancreatitis fibrosis, alcoholic fatty liver disease, bronchitis, pneumonia, lumbar spondylitis, and vasculitis.

[0081] Preferably, the gout is selected from acute gouty arthritis, intermittent gout, chronic gout, hyperuricemia, secondary gout, gouty nephropathy, and urinary tract stones.

[0082] Preferably, the respiratory disease is selected from airway obstructive diseases, more preferably bronchial asthma, allergic asthma, intrinsic asthma, exogenous asthma, exercise-induced asthma, and drug-induced asthma.

[0083] Preferably, the kidney disease is selected from primary glomerular disease, secondary glomerular disease, interstitial nephritis, tubular disease, renal vascular disease, hereditary kidney disease, acute kidney injury caused by ischemia-reperfusion, acute kidney injury caused by sepsis, drug-induced acute kidney injury, contrast agent-induced acute kidney injury, and chronic renal failure; more preferably, the primary glomerular disease is selected from focal segmental glomerulosclerosis, crescentic nephritis, minimal change disease, IgA nephropathy, and membranous nephropathy; more preferably, the secondary glomerular disease is selected from ANCA-associated vasculitis, hypertensive nephropathy, diabetic nephropathy, hepatitis B-related nephritis, lupus nephritis, purpura nephritis, hyperuricemia, and lipoprotein nephropathy; more preferably, the hereditary kidney disease is selected from polycystic kidney disease; more preferably, the drug-induced acute kidney injury is selected from cisplatin-induced acute kidney injury, folic acid-induced acute kidney injury, and aristolochic acid-induced acute kidney injury.

[0084] Preferably, the cancer is selected from prostate cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, intestinal cancer, colon cancer, gastric cancer, skin cancer, brain tumor, leukemia, lymphoma.

[0085] Preferably, the pain is selected from headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone pain, pain caused by cancer and tumor invasion, and neuropathic pain syndrome.

[0086] Preferably, the central nervous system disease is selected from Alzheimer's disease, Parkinson's syndrome, epilepsy, multiple sclerosis and other demyelinating syndromes, cerebral atherosclerosis, and myasthenia gravis.

[0087] Preferably, the lupus erythematosus is systemic lupus erythematosus.

[0088] The present application also provides the use of the compound, its salt, its isotope compound, its stereoisomer or its hydrate in the method of treating diseases related to P2X7 receptor.

[0089] Preferably, the diseases associated with the P2X7 receptor are drugs for inflammation and inflammation-related diseases, kidney diseases, respiratory diseases, cancer, pain, central nervous system diseases, radiation brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis, and allergic asthma.

[0090] Preferably, the inflammation and inflammation-related diseases are selected from neurological inflammation, arthritis, colitis, pancreatitis fibrosis, alcoholic fatty liver disease, bronchitis, pneumonia, lumbar spondylitis, and vasculitis.

[0091] Preferably, the gout is selected from acute gouty arthritis, intermittent gout, chronic gout, hyperuricemia, secondary gout, gouty nephropathy, and urinary tract stones.

[0092] Preferably, the respiratory disease is selected from airway obstructive diseases, more preferably bronchial asthma, allergic asthma, intrinsic asthma, exogenous asthma, exercise-induced asthma, and drug-induced asthma.

[0093] Preferably, the kidney disease is selected from primary glomerular disease, secondary glomerular disease, interstitial nephritis, tubular disease, renal vascular disease, hereditary kidney disease, acute kidney injury caused by ischemia-reperfusion, acute kidney injury caused by sepsis, drug-induced acute kidney injury, contrast agent-induced acute kidney injury, and chronic renal failure; more preferably, the primary glomerular disease is selected from focal segmental glomerulosclerosis, crescentic nephritis, minimal change disease, IgA nephropathy, and membranous nephropathy; more preferably, the secondary glomerular disease is selected from ANCA-associated vasculitis, hypertensive nephropathy, diabetic nephropathy, hepatitis B-related nephritis, lupus nephritis, purpura nephritis, hyperuricemia, and lipoprotein nephropathy; more preferably, the hereditary kidney disease is selected from polycystic kidney disease; more preferably, the drug-induced acute kidney injury is selected from cisplatin-induced acute kidney injury, folic acid-induced acute kidney injury, and aristolochic acid-induced acute kidney injury.

[0094] Preferably, the cancer is selected from prostate cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, intestinal cancer, colon cancer, gastric cancer, skin cancer, brain tumor, leukemia, lymphoma.

[0095] Preferably, the pain is selected from headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone pain, pain caused by cancer and tumor invasion, and neuropathic pain syndrome.

[0096] Preferably, the central nervous system disease is selected from Alzheimer's disease, Parkinson's syndrome, epilepsy, multiple sclerosis and other demyelinating syndromes, cerebral atherosclerosis, and myasthenia gravis.

[0097] Preferably, the lupus erythematosus is systemic lupus erythematosus.

[0098] Definitions of terms used in this application: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, they should be given the meaning that a person skilled in the art would give them based on the disclosure and context.

[0099] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, the prefix C a～b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. For example, C 1～8 Alkyl refers to a straight-chain or branched alkyl group containing 1 to 8 carbon atoms, such as methyl, ethyl, propyl, butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, heptyl, octyl, and isomers of hexyl, heptyl, and octyl.

[0100] As used herein, "aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group with a conjugated π electron system. The aryl ring may be fused to other cyclic groups (including saturated and unsaturated rings), but cannot contain heteroatoms such as nitrogen, oxygen, or sulfur. The point of attachment to the parent group must be on a carbon atom in the ring with a conjugated π electron system. Examples include phenyl, naphthyl, benzyl, and biphenyl. Aryl groups may be substituted or unsubstituted.

[0101] As used herein, "heteroaryl" refers to a heteroaromatic group containing one to two or more heteroatoms. The heteroatoms referred to herein include nitrogen, oxygen, sulfur, phosphorus, and the like. Examples include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, and tetrazolyl. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. The heteroaryl group may be optionally substituted or unsubstituted.

[0102] In this application, an "isotopic compound" refers to a compound in which one or more atoms are replaced by their corresponding isotopes. For example, a compound in which one or more hydrogens (H) are replaced by deuterium (D) or tritium (T); or a compound in which one or more carbons (C12) are replaced by carbons (C11) or (C13).

[0103] In this application, "pharmaceutically acceptable excipients" refer to auxiliary materials widely used in the field of drug production. These excipients should be non-toxic, do not interfere with or impair the efficacy of the active ingredients of the drug, and can be flexibly selected according to the specific dosage form of the drug. These excipients are generally used to improve the physical properties, stability, solubility or absorbability of the drug. The main purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in nature and / or has specific functionality, while helping the active ingredients to dissolve at a desired rate or be effectively absorbed in the body. Pharmaceutically acceptable excipients include, but are not limited to: excipients, which are used to increase the volume or weight of the drug to facilitate formulation molding and processing, such as cocoa butter, suppository wax, vegetable oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil, etc.), alcohols (such as propylene glycol), esters (such as ethyl oleate, ethyl dodecanoate), etc.; binders, which are used to bind drug powder particles together to form tablets with a certain hardness and shape, such as microcrystalline cellulose, hydroxypropyl cellulose, povidone, starch, dextrin, etc.; disintegrants, which are used to quickly disintegrate tablets into fine particles after administration to facilitate drug dissolution and absorption. Lubricants include, for example, cross-linked polyvinylpyrrolidone, pregelatinized starch, silicon dioxide, and cross-linked sodium carboxymethylcellulose. Lubricants are used to reduce friction between drug particles, making tablets easier to compress and form, while also reducing resistance during administration. Examples include magnesium stearate and sodium lauryl sulfate. Coating agents form a thin film on the drug surface to improve its taste, stability, moisture resistance, or control its release rate. Examples include polyacrylic acid resin (used as a coating material for tablets, pills, and granules), hypromellose (used as a water-soluble film coating material for tablets and pills), and sugar coatings (such as those made from sucrose or glucose). Other excipients include colorants (used to color the drug for easier identification), flavorings (used to improve the taste of the drug), sweeteners (used to increase the sweetness of the drug), and preservatives (used to prevent drug deterioration). For example, benzalkonium chloride can be used as a preservative, and fatty acid esters of sorbitol can be used as emulsifiers.

[0104] In the present application, halogen is fluorine, chlorine, bromine or iodine.

[0105] In this application, "alkoxy" is usually represented by RO-, where R represents the alkyl part. It is a type of substituent in organic compound molecules, consisting of an alkyl group and an oxygen atom, such as methyl (CH3-), ethyl (C2H5-), etc.1～8 "Alkoxy" refers to an alkoxy group having 1 to 8 carbon atoms, that is, C1 alkoxy (methoxy) to C8 alkoxy groups are included in this range, such as methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, and octyloxy.

[0106] In this application, "halogenated C 1～8 "Alkoxy" refers to a compound in which one or more hydrogen atoms on the alkyl group in the alkoxy group are replaced by a halogen atom, such as monochloromethoxy, dichloroethoxy, octafluorooctyloxy, etc.

[0107] In this application, "halogenated C 1～8 "Alkyl" refers to a group formed by replacing one or more (such as 1, 2, 3, 4 or 5) hydrogen atoms in an alkyl group (a straight or branched saturated hydrocarbon group with 1 to 8 carbon atoms) with a halogen (fluorine, chlorine, bromine, iodine). Alkyl is a saturated hydrocarbon group in a hydrocarbon molecule, and its general formula is C n H 2n+1 , where n is the number of carbon atoms. In this example, n ranges from 1 to 8. Halogenation refers to the process in which one or more hydrogen atoms in an alkyl group are replaced by a halogen atom. Halogens include fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). Specific examples include: monochloromethyl (CH2Cl), dichloromethyl (CHCl2), trifluoromethyl (CF3), 1,2-dichloroethyl (CHClCH2Cl), trichloroethyl. Other haloalkyl groups include, but are not limited to, monobromoethyl, difluoroethyl, trifluoroethyl, monofluorocyclopropyl, difluorocyclopropyl, etc., formed when some or all of the hydrogen atoms in an alkyl group are replaced by a halogen atom.

[0108] In this application, "3- to 8-membered cycloalkyl" includes saturated and unsaturated cyclic hydrocarbon substituents, and the cyclic hydrocarbon ring atoms do not contain heteroatoms (including but not limited to O, S, or N). For example, 3- to 8-membered cycloalkyl includes saturated cycloalkyl with 3 to 8 ring atoms and unsaturated cycloalkyl with 3 to 8 ring atoms. Examples of 3- to 8-membered cycloalkyl are:

[0109] In this application, "saturated cycloalkyl" refers to a saturated hydrocarbon group with a cyclic structure, in which the carbon atoms are connected by single bonds, and there are no double bonds or triple bonds. The cyclic hydrocarbon has no heteroatoms (including but not limited to O, S or N) in the ring atoms. For example, a 3- to 8-membered saturated cycloalkyl group refers to a saturated cycloalkyl group with 3 to 8 ring atoms, such as: cyclopropyl (C3H5-), cyclobutyl (C4H7-), cyclopentyl (C5H9-), cyclohexyl (C6H 11 -), norbornyl.

[0110] In this application, a "saturated heterocyclic group" refers to a saturated group with a cyclic structure, wherein the ring atoms contain at least one heteroatom (such as nitrogen, oxygen, sulfur, etc.) in addition to carbon atoms, and all bonds within the ring are single bonds, with no unsaturated bonds (such as double bonds or triple bonds). For example, a 3- to 8-membered saturated heterocyclic group refers to a saturated heterocyclic group with 3 to 8 ring atoms. Specific examples include tetrahydrofuranyl, pyrrolidinyl, piperidinyl, tetrahydrothienyl, and morpholinyl.

[0111] In the present application, "fused heterocyclic group" refers to a polycyclic heterocyclic group of 5 to 20 members (more commonly 6 to 14 members, more preferably 7 to 10 members), wherein each ring shares a pair of adjacent carbon atoms with other rings; these rings may contain one or more double bonds, but no ring has a completely conjugated electron system; some or all of the ring atoms in the fused heterocyclic group are heteroatoms (such as nitrogen, oxygen, sulfur, etc.), and the remaining ring atoms are carbon. Depending on the number of member rings, the fused heterocyclic group can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups.

[0112] The compound of Formula I provided herein, as a P2X7 receptor antagonist, not only effectively antagonizes P2X7 receptor activity but also exhibits excellent metabolic stability and pharmacokinetic properties. Compared to the positive control, JNJ47965567, the compound of the present application exhibits significantly improved metabolic stability and pharmacokinetic properties, resulting in significantly better drugability.

[0113] As is well known to those skilled in the art, P2X7 receptor antagonists can be used to treat inflammation and inflammation-related diseases, kidney diseases, gout, respiratory diseases, cancer, pain, central nervous system diseases, radiation-induced brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis, allergic asthma, and the like. Therefore, the compound represented by Formula I, as a P2X7 receptor antagonist, can be used to prepare a medicament for treating inflammation and inflammation-related diseases, kidney diseases, respiratory diseases, cancer, pain, central nervous system diseases, radiation-induced brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis, allergic asthma, and the like, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0114] Figure 1 is a metabolic phenotype study of compound A28 of the present application; wherein the control group represents the metabolic rate of the compound without the addition of any CYP450 enzyme inhibitor; the relative inhibition rate is obtained by calculating the ratio of the metabolic rate of the CYP enzyme experimental group to that of the control group;

[0115] FIG2 shows the effect of compound A27 of the present application on hP2X7-HEK293 current activated by Bz-ATP;

[0116] Figure 3 shows the effect of compound A27 of the present application on ATP-induced mitochondrial damage; wherein, (A) cells were treated with ATP (3 mM) for 1 hour and then stained with MitoTracker (red), which marks mitochondrial membrane potential; (B) cells were stained with MitoSOX (red), which marks mitochondrial reactive oxygen species; the degree of mitochondrial damage was analyzed using a fluorescence microscope (NIKON Eclipse Ts2R / FL inverted microscope), the scale bar represents 100 μm, and the ratio of MitoTracker to MitoSOX was calculated; statistical differences were analyzed using one-way ANOVA: ### indicates successful modeling, ***P<0.001;

[0117] Figure 4 shows that compound A27 of the present application alleviates renal fibrosis in UUO mice; wherein, (A) animal experiment schematic diagram; (B) H&E staining of renal tissue of mice in each group; (C) renal tubular injury score of mice in each group (n=6); (D) Masson staining and α-SMA immunohistochemical staining of renal tissue of mice in each group; (E) collagen deposition score of renal tissue of mice in each group (n=6); (F) positive area of ​​α-SMA immunohistochemical staining of renal tissue of mice in each group (n=6); (G) Fn1 mRNA level of renal tissue of mice in each group (H) Acta2 mRNA levels in renal tissues of mice in each group (n=6); (I) Col1a1 mRNA levels in renal tissues of mice in each group (n=6); (J) Col4a1 mRNA levels in renal tissues of mice in each group (n=6); Data are expressed as mean ± SD; Abbreviations: po, oral administration; ip, intraperitoneal injection; biw., twice a week; ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05, ns, not significant;

[0118] Figure 5 shows that compound A27 of the present application alleviates adenine-induced renal fibrosis in CKD mice; (A) Schematic diagram of the animal experiment; (B) H&E staining of renal tissues of mice in each group; (C) renal tubular injury scores of mice in each group (n=6); (D) blood urea nitrogen levels of mice in each group (n=6); (E) serum creatinine levels of mice in each group; (F) Masson staining and α-SMA immunohistochemical staining of renal tissues of mice in each group; (G) collagen deposition score of renal tissues of mice in each group (n=6); (H) positive area of ​​α-SMA immunohistochemical staining of renal tissues of mice in each group (n=6); (I) Fn1, Col1a1, Col4a1 and A in renal tissues of mice in each group The levels of cta2 mRNA (n=6, ****P<0.0001, adenine group vs. blank control group; ####P<0.0001, adenine A27 treatment group vs. adenine group; $$$P<0.001, $$P<0.01, $P<0.05, adenine A27 treatment group vs. adenine A839977 treatment group) were significantly higher in the renal tissues of the mice in each group. (J) The levels of Fibronectin, Collagen I, Collagen VI, and α-SMA proteins in the renal tissues of the mice in each group were significantly higher in the renal tissues of the mice in each group (n=6, ****P<0.0001, adenine group vs. blank control group; ####P<0.0001, adenine A27 treatment group vs. Adenine A27 treatment group vs. adenine group; $$$$P<0.0001, $$P<0.01, $P<0.05, adenine A27 treatment group vs. adenine A839977 treatment group); (K) Il1b, Nlrp3, Gsdmd and Casp1 mRNA levels in kidney tissues of mice in each group (n=6, ****P<0.0001, adenine group vs. blank control group; ####P<0.0001, adenine A27 treatment group vs. adenine group; $$$P<0.001, $P<0.05, adenine A27 treatment group vs. adenine A839977 treatment group); (L) Mice in each group Renal tissue NLRP3, cleaved Caspase-1 (p20), GSDMD-N, and IL-1β protein levels (n=6, ****P<0.0001, adenine group vs. blank control group; ####P<0.0001, adenine A27-treated group vs. adenine group; $$$$P<0.0001, $$P<0.01, adenine A27-treated group vs. adenine A839977-treated group); data are expressed as mean ± SD; Abbreviations: po, oral; ip, intraperitoneal; biw., twice weekly; ****P<0.0001, ***P<0.001;

[0119] Figure 6 shows the effect of compound A27 of the present application on gouty arthritis mice; (A) is a representative image of the mouse ankle joint, clearly depicting the differences in the appearance of the ankle joints of mice in different experimental groups; (B) is the quantitative data of the mouse ankle swelling index, which intuitively reflects the degree of ankle swelling in each group of mice; (C) is the measurement result of IL-1β content in mouse serum, revealing the changes in the levels of inflammatory factors among different experimental groups; (D) is the measurement result of TNF-α content in mouse serum, further revealing the differences in inflammatory factors among different experimental groups; all data are presented as mean ± standard deviation (SD) and analyzed using one-way ANOVA; sample size (n) = 4; in the statistical results, ### indicates successful model establishment, **P<0.01 indicates a statistically significant difference compared with the control group, and ***P<0.001 indicates an extremely significant difference compared with the control group. DETAILED DESCRIPTION

[0120] The following is a detailed description of the above content of this application through specific implementation methods in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of this application to the following examples. Obviously, based on the above content of this application, in accordance with common technical knowledge and customary means in this field, various other forms of modification, replacement, or change can be made without departing from the basic technical concept of this application. All technologies implemented based on the above content of this application fall within the scope of this application.

[0121] The raw materials and equipment used in this application are all known products and are obtained by purchasing commercially available products.

[0122] To facilitate the subsequent description of the synthetic routes and methods of the examples, the abbreviations of some of the raw materials or reagents used in the examples are listed in Table 1.

[0123] Table 1 Abbreviations of some raw materials or reagents used in the examples

[0124] Example 1 Preparation of N-((1-(5-bromopyridin-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-2-(4-(methylsulfonyl)phenyl)thiazole-5-carboxamide (A1)

[0125] Step 1: Preparation of Intermediate 1

[0126] 2-Fluoro-5-bromopyridine (1 eq.), hydrazine hydrate (5 eq.), and ethanol (5 ml) were added to the reaction flask and stirred in an oil bath at 60°C for 1.5 h, monitoring the reaction by TLC. After completion, the reaction solution was cooled to room temperature. A white solid precipitated, filtered, washed with ether, and dried. The white solid was diluted with 1 ml of water without purification and 3 ml of glacial acetic acid was added. Aqueous NaNO₂ (5 eq. NaNO₂ dissolved in 10 ml of water) was slowly added dropwise with stirring at room temperature. After the addition was complete, the mixture was stirred at room temperature for 2 h, monitoring the reaction by TLC. After completion of the reaction, saturated sodium carbonate solution was added to the reaction system to adjust the pH to neutral. An appropriate amount of water was added, and the mixture was extracted three times with a mixed organic solvent system of EA:PE = 1:1. The organic phases were combined and distilled under reduced pressure at low temperature to obtain a white solid (Intermediate 1).

[0127] 1 HNMR (400MHz, DMSO-d6) δ9.38(d,J=7.3Hz,1H),8.52(d,J=2.0Hz,1H),7.56(dd,J=7.3,2.1Hz,1H).

[0128] Step 2: Preparation of Intermediate 2

[0129] 2-bromothiazole-5-carboxylic acid (1 eq.) and 5 ml of DCM were added to the reaction flask, and a drop of DMF was added as a catalyst. The reaction was stirred at room temperature for 4 hours. The reaction solution was spin-dried to obtain an intermediate of 2-bromothiazole-5-carboxylic acid chloride. A clean reaction flask was added, triethylamine (2.1 eq.), propargylamine (1 eq.) and 3 ml of DMF were added, and the prepared benzoyl chloride intermediate was dissolved in 2 ml of DMF. The mixture was then added dropwise to the reaction system under stirring at room temperature. The reaction was stirred at room temperature for 0.5 hours, and the reaction was monitored by TLC during this period. After the reaction was completed, a large amount of water was added, and a white solid was precipitated. The product was filtered, washed with ether, and dried to obtain a white solid (intermediate 2).

[0130] 1 HNMR (400MHz, DMSO-d6) δ9.32–9.26(m,1H),8.25(s,1H),4.05(dd,J=5.5,2.7Hz,2H),3.21(t,J=2.4Hz,1H).

[0131] Step 3: Preparation of Intermediate 3

[0132] Intermediate 2 (1 eq.), 4-methanesulfonylphenylboronic acid (1.5 eq.), tetrakis(triphenylphosphine)palladium (0.1 eq.), and sodium carbonate (2 eq.) were dissolved in a mixture of 5 mL of dioxane and 1 mL of water. Under nitrogen, the reaction mixture was stirred at 85°C for 8 hours and monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to obtain Intermediate 3.

[0133] 1 H NMR(400MHz, DMSO-d6)δ9.32(t,J=5.5Hz,1H),8.56(d,J=1.6Hz,1H),8.27–8.23(m,2H),8.06(d d,J=8.5,1.7Hz,2H),4.09(dd,J=5.7,2.7Hz,2H),3.28(d,J=1.6Hz,3H),3.21(t,J=2.4Hz,1H).

[0134] Step 4: Preparation of Compound A1

[0135] To a dry three-necked flask, add Intermediate 1 (1.5 eq.), Intermediate 3 (1 eq.), sodium L-ascorbate (5 eq.), copper sulfate pentahydrate (1.5 eq.), and 5 ml of DMF. Under nitrogen, stir at 35°C and react overnight, monitoring the reaction by TLC. After completion, add a large amount of water and extract three times with dichloromethane. The organic phase is collected, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to obtain product A1.

[0136] 1 H NMR (400MHz, DMSO-d6) δ9.47(t,J=5.6Hz,1H),8.75(d,J=2.4Hz,1H),8.74(s,1H),8.60(s,1H),8.36(dd,J=8.7,2 .4Hz,1H),8.27–8.23(m,2H),8.09(d,J=8.8Hz,1H),8.08–8.04(m,2H),4.64(d,J=5.6Hz,2H),3.28(s,3H).Exact mass calcd for C 19 H 15 Br N6O3S2Na,540.9728,[M+Na]+:540.9723.

[0137] Examples 2 to 11

[0138] Referring to the method of Example 1, compounds A2-A11 can be prepared by changing the raw materials for preparing intermediate 1.

[0139] Example 12 Preparation of N-((1-(4-bromopyridin-2-yl)-1H-1,2,3-triazol-4-yl)methyl)-2-(4-(methylsulfonyl)phenyl)thiazole-4-carboxamide (A12)

[0140] Step 1: Preparation of intermediate 4

[0141] 2-Fluoro-4-bromopyridine (1 eq.), hydrazine hydrate (5 eq.), and ethanol (5 ml) were added to the reaction flask and stirred in an oil bath at 60°C for 1.5 h, during which the reaction was monitored by TLC. After completion of the reaction, the reaction solution was cooled to room temperature, and a white solid precipitated. This solid was filtered, washed with ether, and dried. The white solid was diluted with 1 ml of water without purification, and 3 ml of glacial acetic acid was added. Aqueous NaNO2 (5 eq. NaNO2 dissolved in 10 ml of water) was slowly added dropwise with stirring at room temperature. After the addition was complete, the solution was stirred at room temperature for 2 h, during which the reaction was monitored by TLC. After completion of the reaction, saturated sodium carbonate solution was added to the reaction system to adjust the pH to neutral. An appropriate amount of water was added, and the solution was extracted three times with a mixed organic solvent system of EA:PE = 1:1. The organic phases were combined and distilled under reduced pressure at low temperature to obtain a white solid (Intermediate 4).

[0142] 1 HNMR (400MHz, DMSO-d6) δ9.29(d,J=7.2Hz,1H),8.66(d,J=1.9Hz,1H),7.63(dd,J=7.2,1.9Hz,1H).

[0143] Step 2: Preparation of Intermediate 5

[0144] 2-bromo-4-thiazolecarboxylic acid (1 eq.) and 5 ml of DCM were added to the reaction flask, and a drop of DMF was added as a catalyst. The reaction was stirred at room temperature for 4 hours. The reaction solution was spin-dried to obtain an intermediate of 2-bromothiazole-5-carboxylic acid chloride. A clean reaction flask was added, triethylamine (2.1 eq.), propargylamine (1 eq.) and 3 ml of DMF were added, and the prepared benzoyl chloride intermediate was dissolved in 2 ml of DMF. The solution was then added dropwise to the reaction system under stirring at room temperature, and the reaction was stirred at room temperature for 0.5 hours. During this time, the reaction was monitored by TLC. After the reaction was completed, a large amount of water was added, and a white solid was precipitated. The product was filtered, washed with ether, and dried to obtain a white solid (intermediate 5).

[0145] 1HNMR (400MHz, DMSO-d6) δ8.92(d,J=6.1Hz,1H),8.31(dd,J=13.9,1.4Hz,1H),3.99(dt,J=5.9,1.8Hz,2H),3.11–3.03(m,1H).

[0146] Step 3: Preparation of Intermediate 6

[0147] Intermediate 5 (1 eq.), 4-methanesulfonylphenylboronic acid (1.5 eq.), tetrakis(triphenylphosphine)palladium (0.1 eq.), and sodium carbonate (2 eq.) were dissolved in a mixture of 5 mL of dioxane and 1 mL of water. Under nitrogen, the reaction mixture was stirred at 85°C for 8 hours and monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to obtain Intermediate 6.

[0148] 1 HNMR(400MHz,DMSO-d6)δ9.07(t,J=6.0Hz,1H),8.47(d,J=1.6Hz,1H),8.33(dd,J=8.4,1.7Hz,2 H),8.08(dd,J=8.4,1.7Hz,2H),4.10–4.05(m,2H),3.29(d,J=1.5Hz,3H),3.13(q,J=2.2Hz,1H).

[0149] Step 4: Preparation of Compound A12

[0150] To a dry three-necked flask, add Intermediate 4 (1.5 eq.), Intermediate 6 (1 eq.), sodium L-ascorbate (5 eq.), copper sulfate pentahydrate (1.5 eq.), and 5 ml of DMF. Under nitrogen, stir at 35°C and react overnight, monitoring the reaction by TLC. After completion, add a large amount of water and extract three times with dichloromethane. The organic phase is collected, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to obtain product A12.

[0151] 1 HNMR(400MHz,DMSO-d6)δ9.29(t,J=6.0Hz,1H),8.69(s,1H),8.48(d,J=5.3Hz,2H),8.35–8.32(m,2H),8 .31(d,J=1.7Hz,1H),8.09–8.06(m,2H),7.81(dd,J=5.4,1.7Hz,1H),4.67(d,J=6.0Hz,2H),3.29(s,3H).

[0152] 13 C NMR(100MHz,DMSO-d6)δ165.16,160.41,150.75,150.14,149.09,146.29,142.16,136.64,13 4.71,127.95,127.95,127.24,127.24,127.24,126.03,120.50,116.51,43.37,34.37.Exact mass calcd forC 19 H 15 BrN6O3S2Na,540.9728,[M+Na]+:540.9727.

[0153] Examples 13 to 24

[0154] Referring to the method of Example 12, compounds A13-A24 can be prepared by changing the raw materials for preparing intermediate 4.

[0155] Example 25 Preparation of (S)-N-(1-(1-(4-trifluoromethylpyridin-2-yl)-1H-1,2,3-triazol-4-yl)ethyl)-2-(4-(methylsulfonyl)phenyl)thiazole-4-carboxamide (A25)

[0156] Step 1: Preparation of intermediate 7

[0157] 2-Fluoro-4-trifluoromethylpyridine (1 eq.), hydrazine hydrate (5 eq.), and ethanol (5 ml) were added to the reaction flask and stirred in an oil bath at 60°C for 1.5 h, with TLC monitoring. After completion, the reaction solution was cooled to room temperature, and a white solid precipitated. This solid was filtered, washed with ether, and dried. The white solid was diluted with 1 ml of water without purification, and 3 ml of glacial acetic acid was added. Aqueous NaNO₂ (5 eq. NaNO₂ dissolved in 10 ml of water) was slowly added dropwise with stirring at room temperature. After the addition was complete, the mixture was stirred at room temperature for 2 h, with TLC monitoring. After completion, the pH was adjusted to neutral by adding saturated sodium carbonate solution. Water was added as needed, and the mixture was extracted three times with a 1:1 organic solvent mixture of EA:PE. The organic phases were combined, the solvent removed by vacuum distillation, and purified by column chromatography to yield a colorless, transparent oil (Intermediate 7).

[0158] 1H NMR (400MHz, DMSO-d6) δ9.57 (dt, J=7.2, 0.9Hz, 1H), 8.90 (dq, J=1.9, 1.1Hz, 1H), 7.77 (dd, J=7.2, 1.8Hz, 1H).

[0159] Step 2: Preparation of Intermediate 8

[0160] To a dry three-necked flask, add Intermediate 7 (1.5 eq.), tert-butyl (S)-but-3-yn-2-ylcarbamate (1 eq.), sodium L-ascorbate (5 eq.), copper sulfate pentahydrate (1.5 eq.), and 5 ml of DMF. Under nitrogen, stir at 35°C and react overnight. Monitor the reaction by TLC. After completion, add a large amount of water and extract three times with dichloromethane. The organic phase is collected, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to yield Intermediate 8.

[0161] 1 H NMR (400MHz, DMSO-d6) δ8.88(d,J=5.2Hz,1H),8.65(s,1H),8.34(s,1H),7.93(dd,J=5.2,1.6H z,1H),7.41(d,J=8.4Hz,1H),4.93–4.81(m,1H),1.45(d,J=7.0Hz,3H),1.38(d,J=4.6Hz,9H).

[0162] Step 3: Preparation of Intermediate 9

[0163] To a reaction flask, add intermediate 8 (1 eq.) and 5 ml of DCM, then dropwise add 2 ml of trifluoroacetic acid. Stir the reaction at room temperature while monitoring the reaction by TLC. After completion, adjust the pH to 8 with 3 M NaOH solution. Add water to the reaction system and extract three times with ethyl acetate. The organic phase is collected and concentrated under reduced pressure to remove the solvent to yield intermediate 9. Use it directly in the next step without purification.

[0164] 1 H NMR (400MHz, DMSO-d6) δ8.89(d,J=5.2Hz,1H),8.71(s,1H),8.35(s,1H),7.93(dd,J=5.2,1.6Hz,1H),4.16(d,J=6.8Hz,1H),1.39(d,J=6.7Hz,3H).

[0165] Step 4: Preparation of Intermediate 10

[0166] 2-Bromo-4-thiazolecarboxylic acid (1 eq.) and 5 ml of DCM were added to a reaction flask, and a drop of DMF was added as a catalyst. The reaction was stirred at room temperature for 4 hours, and the reaction solution was spin-dried to obtain an intermediate of 2-bromothiazole-4-carboxylic acid chloride. Separately, a clean reaction flask was added with triethylamine (2.1 eq.), intermediate 9 (1 eq.), and 3 ml of DMF. The prepared acid chloride intermediate was dissolved in 2 ml of DMF and added dropwise to the reaction system under stirring at room temperature. The reaction was stirred at room temperature for 0.5 hours, and the reaction was monitored by TLC during this period. After the reaction was completed, a large amount of water was added to precipitate a white solid, which was filtered, washed with ether, and dried to obtain a white solid (intermediate 10).

[0167] 1 HNMR(400MHz,DMSO-d6)δ8.91–8.87(m,2H),8.79(s,1H),8.35(d,J=1.5Hz,1H),8 .34(s,1H),7.95(dd,J=5.2,1.6Hz,1H),5.44–5.37(m,1H),1.61(d,J=7.0Hz,3H).

[0168] Step 5: Preparation of Compound A25

[0169] Intermediate 10 (1 eq.), 4-methanesulfonylphenylboronic acid (1.5 eq.), tetrakis(triphenylphosphine)palladium (0.1 eq.), and sodium carbonate (2 eq.) were dissolved in a mixture of 5 mL of dioxane and 1 mL of water. Under nitrogen, the reaction mixture was stirred at 85°C for 8 hours and monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to obtain compound A25.

[0170] 1 H NMR (400MHz, DMSO-d6) δ8.99(d,J=8.6Hz,1H),8.89(d,J=5.2Hz,1H),8.84(s,1H),8.49(s,1H),8.38–8.32(m ,3H),8.10–8.05(m,2H),7.95(d,J=5.2Hz,1H),5.51–5.44(m,1H),3.29(s,3H),1.68(d,J=7.0Hz,3H).Exact mass calcd for C 21 H 17 F3N6O3S2Na,545.0653,[M+Na]+:545.0653.

[0171] Example 27 Preparation of (S)-2-(4-(methylsulfonyl)phenyl)-N-(1-(1-(4-trifluoromethylpyridin-2-yl)-1H-1,2,3-triazol-4-yl)ethylthiazole-5-carboxamide (A27)

[0172] Step 1: Preparation of intermediate 11

[0173] 2-bromothiazole-5-carboxylic acid (1 eq.) and 5 ml of DCM were added to the reaction flask, and a drop of DMF was added as a catalyst. The reaction was stirred at room temperature for 4 hours. The reaction solution was spin-dried to obtain an intermediate of 2-bromothiazole-5-carboxylic acid chloride. A clean reaction flask was added, triethylamine (2.1 eq.), intermediate 9 (1 eq.) and 3 ml of DMF were added. The prepared acyl chloride intermediate was dissolved with 2 ml of DMF and then added dropwise to the reaction system under stirring at room temperature. The reaction was stirred at room temperature for 0.5 hours, and the reaction was monitored by TLC during this period. After the reaction was completed, a large amount of water was added, and a white solid was separated out. The product was filtered, washed with ether, and dried to obtain a white solid (intermediate 11).

[0174] 1 H NMR (400MHz, DMSO-d6) δ9.24(d,J=7.9Hz,1H),8.89(d,J=5.2Hz,1H),8.84(s,1H),8.36( s,1H),8.32(s,1H),7.95(d,J=5.1Hz,1H),5.35(p,J=7.1Hz,1H),1.62(d,J=7.0Hz,3H).

[0175] Step 2: Preparation of Compound A27

[0176] Intermediate 11 (1 eq.), 4-methanesulfonylphenylboronic acid (1.5 eq.), tetrakistriphenylphosphine palladium (0.1 eq.), and sodium carbonate (2 eq.) were dissolved in a mixture of 5 mL of dioxane and 1 mL of water. Under nitrogen, the reaction mixture was stirred at 85°C for 8 hours and monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and purified by silica gel column chromatography to obtain compound A27.

[0177] 1H NMR (400MHz, DMSO-d6) δ9.35(d,J=7.9Hz,1H),8.89(d,J=5.2Hz,1H),8.86(s,1H),8.67(s,1H),8.36(s,1H),8.27–8. 23(m,2H),8.08–8.04(m,2H),7.95(dd,J=5.1,1.6Hz,1H),5.41(p,J=7.1Hz,1H),3.28(s,3H),1.66(d,J=7.0Hz,3H).

[0178] 13 C NMR(100MHz,DMSO-d6)δ168.05,158.96,151.03,150.61,149.30,144.71,142.35,139.90,136.95,136.72, 128.11,128.11,127.29,127.29,122.33(q,J=272.1),119.85,119.66,109.64,43.38,41.78,20.26.Exact mass calcd for C 21 H 18 F3N6O3S2,523.0834,[M+H] + :523.0830.

[0179] Examples 26-32

[0180] Referring to the methods of Example 25 and Example 27, compounds A26 and A28-A32 can be prepared by changing the raw materials of intermediate 7 and intermediate 9 or 11.

[0181] The beneficial effects of the present application are demonstrated below through specific test examples.

[0182] Test Example 1: Antagonistic activity test of compounds on P2X7 receptors

[0183] 1. Experimental Methods

[0184] As is well known to those skilled in the art, the P2X7 receptor (P2X7R) is primarily regulated by ATP. High ATP concentrations of 0.5 to 1 mM and above can stimulate P2X7R activation, leading to the opening of the receptor pore. The open "macropore" allows the uptake of hydrophilic solutes with molecular weights up to 900 Da (such as ethidium bromide, YO-Pro, propidium iodide (PI), or Lucifer Yellow), which are otherwise impermeable to the intracellular environment. In this experiment, PI was used as a fluorescent dye to conduct PI dye uptake experiments.

[0185] By lentiviral transfection, HEK293 (human embryonic kidney 293) cells (mP2X7-HEK293 and hP2X7-HEK293) were constructed to stably express human and mouse P2X7R. An in vitro cell-based assay was established using ATP-induced P2X7R activation and simultaneous labeling with fluorescent dyes to test the antagonistic activity of drugs against hP2X7R (human P2X7R) and mP2X7R (mouse P2X7R).

[0186] Materials: PI, ATP, DMEM culture medium.

[0187] Test drugs: Compounds A1-A32, with the known P2X7R antagonist JNJ47965567 as a positive control.

[0188] Preparation of drug solution:

[0189] The compound was prepared into a 10 mmol / L stock solution using 100% DMSO;

[0190] PI was prepared into a 1 mg / ml stock solution using sterile PBS;

[0191] ATP was prepared into a 600 mM stock solution in sterile PBS.

[0192] Assay Method: mP2X7R-HEK293 and hP2X7R-HEK293 cells (2×105 cells / mL, 100 μL / well) were seeded into 96-well black, transparent flat-bottom microplates and incubated in an incubator (37°C, 25% CO2) for 24 hours. Compounds, PI, and ATP were diluted to their active concentrations in DMEM medium and mixed and added to the plates (100 μL / well). Incubated in the dark for 2 hours, the medium was aspirated and pre-chilled PBS (100 μL / well) was added. Fluorescence was immediately measured using a microplate reader (Ex / Em = 535 / 617 nM). The active concentrations of PI and ATP were 0.05 g / mL and 3 mM, respectively. The raw data for each well was read and recorded, and the raw data were converted accordingly to calculate the antagonistic activity of each compound against hP2X7R and mP2X7R.

[0193] 2. Experimental Results

[0194] The results are shown in Tables 2 and 3. The effective concentration of the compounds in Table 2 was 1 μM, and the effective concentration of the compounds in Table 3 was 100 nM. Antagonism rates > 80% were indicated as "+++," 50% < antagonism < 80% was indicated as "++," and antagonism rates < 50% were indicated as "+."

[0195] Table 2 Antagonistic activity of compounds at 1 μM against hP2X7R and mP2X7R

[0196] As can be seen from Table 2 above, the compounds of the present application all have antagonistic activity on P2X7R at 1 μM, among which compounds A9, A12, A19 and A25 all showed good activity on P2X7R in humans and mice, and the antagonistic effect at this concentration was comparable to that of the positive control JNJ47965567.

[0197] Table 3 Antagonistic activity of compounds at 100 nM against mP2X7R and hP2X7R

[0198] As can be seen from Table 3 above, the antagonistic effects of compounds A9, A12, A19 and A25 of the present application on P2X7R at low concentrations (100 nM) are still comparable to those of the positive control JNJ47965567.

[0199] The above results indicate that the compounds provided in the present application can effectively antagonize the activity of human and mouse P2X7 receptors and can be used to prepare P2X7 receptor antagonists.

[0200] Test Example 2 Liver microsome metabolic stability test

[0201] 1. Experimental Methods

[0202] Test drugs: Compounds A8, A9, A12, A19, A20, A21, A22, A24, A25, A26, A27, A28, A29, A30, A31, A32, with the known P2X7R antagonist JNJ47965567 as a positive control.

[0203] Test method: The compound is incubated in vitro with human or mouse liver microsome solution at 37°C. The concentration of the compound at the 0-min incubation time point is set as 100%. The concentrations at other incubation time points are compared with this concentration to obtain the residual percentage. The natural logarithm of the residual percentage at each time point is linearly regressed against the incubation time to obtain the slope k. The half-life (T) can be calculated according to the following formula: 1 / 2 ) and clearance (CL). T 1 / 2 =-0.693 / k; CL = [0.693 / T 1 / 2 ]×[incubation medium volume (ml) / microsome mass (mg)].

[0204] 2. Experimental Results

[0205] Table 4 Metabolic stability test results of the compounds Note: h and m represent human and mouse species, respectively.

[0206] As can be seen from Table 4, compared with the positive control JNJ47965567, the half-life of the compound of the present application is significantly prolonged and the clearance rate is significantly reduced, indicating that the metabolic stability of the compound of the present application is significantly improved compared with the positive control JNJ47965567.

[0207] Test Example 3 Pharmacokinetic Test

[0208] 1. Experimental Methods

[0209] Test drugs: Compounds A12, A19, A25, A27, and A29, with the known P2X7R antagonist JNJ47965567 as a positive control.

[0210] Test Methods: Male BALB / c mice (18-22 g) were randomly divided into two groups (n=3) and administered intravenously (10 mg / kg) or orally (10 mg / kg). All animal experiments were conducted in accordance with the standard guidelines for the care and use of laboratory animals. The compound was administered at 1.0 mg / mL in normal saline. Eye blood was collected 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 24 hours, and 48 hours after administration. All blood samples were centrifuged and quantitatively analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using DAS2.0.

[0211] Table 5 Intravenous pharmacokinetic test results of the compounds

[0212] Table 6 Oral administration pharmacokinetic test results of the compounds

[0213] As can be seen from Tables 5 and 6, compared with the positive control JNJ47965567, the pharmacokinetic properties of the compound of the present application are significantly improved, the half-life is significantly prolonged, the clearance rate is significantly reduced, and the in vivo exposure is significantly increased.

[0214] The experimental results of Test Example 2 and Test Example 3 show that compared with the positive control JNJ47965567, the compound of the present application has better drugability.

[0215] Test Example 4 Plasma protein binding rate experiment

[0216] The plasma protein binding test was performed using a plasma matrix containing 10 μM of the drug. The dialysis chamber of the dialysis plate was filled with phosphate buffered saline (PBS, pH 7.4), and an equal amount of drug-containing plasma matrix was added to the sample side. Analysis was performed after 6 hours of reaction at 37°C. After the reaction was completed, samples were taken from each compartment, treated with acetonitrile containing the internal standard SAHA, and centrifuged. The supernatant was then placed in an injection bottle for LC-MS / MS analysis. The control sample was prepared in the same way, but the dialysis step was omitted, and all experiments were repeated three times to ensure accuracy and reliability. The test results are presented as the percentage of protein binding in plasma. The plasma protein binding rate of compound A28 is 97.87%, the plasma protein binding rate of compound A25 is 98.62%, and the plasma protein binding rate of compound A27 is 99.27%.

[0217] Experimental Example 5 CYP450 metabolic phenotype analysis

[0218] 1 μL of compound A28 (1 μM) was mixed with 5 μL of human liver microsomes and 1 μL of specific CYP450 enzyme inhibitors (ticlopidine hydrochloride as an inhibitor of CYP2C19, α-naphthol as a specific selective inhibitor of CYP1A2, pilocarpine as a specific selective inhibitor of CYP2A6, sodium diethyldithiocarbamate as a specific selective inhibitor of CYP2E1, sulfaphenazole as a specific selective inhibitor of CYP2C9, quinidine as a specific selective inhibitor of CYP2C9). Incubations were performed at 37°C in a total volume of 200 μL with 0.1 M phosphate buffer (pH 7.4), an NADPH regeneration system (1 mM NADP, 5 mM glucose-6-phosphate, 1 U / mL glucose-6-phosphate dehydrogenase, and 3.3 mM magnesium chloride). Each sample was run in triplicate, with a control group without selective inhibitors. After a 2-h incubation, the reaction was terminated by adding 3 volumes of pre-chilled acetonitrile. The residual concentration of the parent drug in the incubation solution was determined by LC-MS / MS.

[0219] The experimental results are shown in Figure 1. Compound A28 was primarily metabolized by CYP2C19, CYP2D6, and CYP2E1 within two hours, with the remaining 84.2%. The experimental results showed that compound A27 was essentially not metabolized within two hours.

[0220] Experimental Example 6 Drug Renal Distribution

[0221] Animal experiments were conducted to investigate the distribution of compound A27 in plasma and kidneys. These experiments were approved by the Animal Experiment Committee of West China Hospital, Sichuan University (IACUC number: 2020192A), and the care and use of experimental animals were carried out in accordance with the regulations. Compound A27 was prepared into a 1 mg / mL solution with 5% DMSO and 95% saline. Balb / c male mice (3 mice at each time point) were gavaged at 10 mg / kg (200 μL injection volume per 20 g mouse body weight). Plasma samples and mouse kidneys were collected at 1 h, 6 h, 24 h, 3 d, 7 d, and 14 d after administration. Plasma samples were centrifuged at 4500 rpm for 10 minutes, and 5 μL of plasma was mixed with 100 μL of acetonitrile containing 20 ng / mL internal standard SAHA and centrifuged at 13000 rpm for 10 minutes. The supernatant was collected again and analyzed by LC-MS / MS (ABSCIEX 5500 Qtrapmass spherometer). Kidney samples were added to deionized water and ground with pre-cooled steel balls (60 Hz, 2 minutes). After grinding, the balls were removed and 20 μL of the homogenate was added to 200 μL of acetonitrile containing 20 ng / mL internal standard SAHA. The mixture was centrifuged at 13,000 rpm for 10 minutes. The supernatant was again transferred to a vial for analysis. Standard concentrations of the test compound were prepared and a standard curve was established. Pharmacokinetic parameters were analyzed using DAS2.0 software.

[0222] Table 7 Drug concentrations in blood and kidneys within one week after single oral administration of A27

[0223] The experimental results are shown in Table 7. After oral administration of 10 mg / kg once, the concentration in the mouse kidney was 373.18 ng / g after 1 week, which is still higher than the IC 90 (337.54 ng / mL)

[0224] Experimental Example 7 Electrophysiological Experiment

[0225] HEK293 cells stably expressing the human P2X7 receptor (gene information: NM_002562.6) were used. These cells were cultured in DMEM supplemented with 10% fetal bovine serum at 37°C in a 5% CO2 atmosphere. Before patch-clamp experiments, cells were digested with 0.25% trypsin-EDTA, seeded onto coverslips in 24-well plates (final volume: 500 μL), and incubated for 18 hours. The patch-clamp experiment involved fabricating microelectrodes using an electrode puller, placing the coverslip in a recording chamber under an inverted microscope, and then inserting the microelectrode into the extracellular fluid to measure electrode resistance (Rpip). The electrode was gently applied to the cell surface, and negative pressure was applied to form a high-resistance seal (GΩ). Fast capacitance compensation was then performed. Further negative pressure was then applied to rupture the cell membrane, establishing a whole-cell recording configuration, and measuring slow capacitance compensation and series resistance as experimental parameters. All electrophysiological experiments were performed at room temperature.

[0226] Whole-cell patch clamp recordings of P2X7 receptor currents utilized gap current mode stimulation at -80 mV. Cells were first treated with 100 μM BZ-ATP, followed by a 1-minute preincubation with the drug, and then the mixed working solution was added at varying concentrations. Peak P2X7 receptor currents induced by drug plus BZ-ATP (100 μM) (A27 + BZ-ATP 100 μM) were compared with peak currents induced by BZ-ATP alone (100 μM) (BZ-ATP 100 μM). The inhibitory percentage of drug concentration was calculated as: % inhibition = [1-(A27 + BZ-ATP 100 μM / BZ-ATP 100 μM)] × 100%. Data are presented as mean ± standard error (SEM). Sustained ATP stimulation increases P2X7 receptor membrane permeability and currents. As shown in FIG2 , compound A27 significantly inhibited hP2X7 receptor currents induced by Bz-ATP in hP2X7-HEK293 cells, with an inhibition rate of 67.99%±0.98% at a concentration of 10 μM.

[0227] Experimental Example 8 Effects on ATP-induced mitochondrial damage

[0228] The P2X7 receptor plays a key role in mitochondrial damage, particularly under ATP stimulation. ATP, as a P2X7 agonist, induces intracellular calcium influx, triggering mitochondrial damage and promoting the production of reactive oxygen species (ROS). Mitochondrial damage can be identified by changes in red oxygen status and membrane potential. When detected using MitoTracker staining, active mitochondria exhibit stronger fluorescence than apoptotic mitochondria. As superoxide levels increase, the fluorescence intensity of MitoSOX increases accordingly. In the experiment, mP2X7R-HEK293 and hP2X7R-HEK293 cells were pretreated with compound A27 for 30 minutes before stimulation with 3 mM ATP. MitoTracker and MitoSOX staining were then added 30 minutes before ATP stimulation. ATP stimulation lasted for 1 hour, after which the cells were washed with ice-cold PBS and observed under a fluorescence microscope. Image analysis was performed using ImageJ software.

[0229] As shown in Figure 3, MitoTracker staining indicated that A27 significantly reduced the proportion of dysfunctional mitochondria in hP2X7R and mP2X7R-HEK293 cells after ATP exposure (Figure 3A). MitoSOX fluorescence results showed that ATP treatment led to increased mitochondrial superoxide levels in cells, indicating increased ROS production, while A27 significantly reduced this level (Figure 3B). These results indicate that compound A27 has a protective effect on mitochondrial function.

[0230] Experimental Example 9 In vivo pharmacodynamic evaluation

[0231] Male C57BL / 6J mice (8–10 weeks old) were purchased from Nanjing GenScript Biotechnology Co., Ltd. All experimental protocols were approved by the Ethics Committee of West China Hospital of Sichuan University (approval number: 20230709001), and mice were randomly divided into groups (n = 6 / group).

[0232] 1) Effect of compound A27 on renal fibrosis in UUO mice

[0233] Construction of the mouse unilateral ureteral obstruction (UUO) model: First, the mouse's back was shaved and disinfected under isoflurane anesthesia. Then, the skin was incised in the left kidney area to expose and ligate the left ureter. The ligation position should be consistent to ensure the comparability of the experiments. Finally, the incision was sutured and the mouse was returned to the cage for recovery. Postoperative monitoring and care were also performed.

[0234] Construction of adenine-induced chronic kidney disease (CKD) model: 8-10 week old male C57BL / 6J mice were fed a diet containing 0.2% adenine (Research Diets, Inc., Brunswick, NJ) (adenine dose of 160 mg / kg / day) and allowed to eat freely for 2 weeks.

[0235] Experimental Methods: Compound A27 was administered orally at doses of 0.3 mg / kg, 1 mg / kg, or 10 mg / kg, once or twice weekly. A839977 (30 μmol / kg, intraperitoneal injection, twice weekly) and pirfenidone (PFD) (250 mg / kg, oral, once daily) were selected as positive controls. After the experiment, the animals were euthanized by cervical dislocation, and blood and kidney tissue samples were collected. Serum was separated by centrifugation at 3000 rpm for 30 minutes. A portion of the kidney tissue sample was snap-frozen in liquid nitrogen and stored at -80°C for Western blot and RT-qPCR analysis. The other portion was fixed in 10% formaldehyde, dehydrated, and embedded in paraffin for H&E, Masson's, and IHC staining. For histological assessment of renal tubular damage and renal fibrosis, kidney sections were scanned at 20x magnification using an Olympus VS200 slide scanner (Tokyo, Japan). Tubular injury was scored as follows: 0, normal; 1, percent injury area <25%; 2, percent injury area 25%-50%; 3, percent injury area 51%-75%; and 4, percent injury area >75%. Masson staining-positive area was assessed using collagen volume fraction (CVF). Immunohistochemical staining-positive area was calculated using ImageJ software (version 1.51, Wayne Rasband, National Institutes of Health).

[0236] As shown in Figure 4, in the unilateral ureteral obstruction (UUO) model (Figure 4A). H&E staining of renal sections and tubular injury scores showed that both doses of A27 reduced tubular injury, characterized by reduced inflammatory cell infiltration and tubular dilation (Figures 4B and C). Masson staining showed that interstitial collagen deposition in the kidneys of UUO mice was increased, which was alleviated after A27 treatment (Figures 4D and E). In addition, immunohistochemistry (IHC) staining of α-SMA showed that both doses of A27 reduced the increase in α-SMA expression in the kidneys of UUO mice (Figures 4D and F). After treatment with both doses of A27, the mRNA levels of profibrotic markers (Fn1, Acta2, Col1a1, and Col4a1) in the kidneys of UUO mice were also reduced (Figures 4G-J). The experimental results showed that lower doses of A27 (0.3 mg / kg and 1 mg / kg) could alleviate renal tubular damage and interstitial collagen deposition in UUO mice, and significantly downregulated the expression of pro-fibrotic markers, among which 1 mg / kg had a more significant effect (Figure 4A-J).

[0237] As shown in Figure 5, A27 significantly alleviated adenine-induced renal injury in CKD mice, reduced serum creatinine and BUN levels, improved renal fibrosis, and downregulated the mRNA and protein levels of pro-fibrotic markers (Figure 5A-J). In addition, A27 also inhibited the activation of the NLRP3 inflammasome, as demonstrated by decreased mRNA levels of Il1b, Nlrp3, Gsdmd, and Casp1, as well as decreased protein levels of NLRP3, activated Caspase-1, GSDMD-N, and activated IL-1β (Figure 5K and L).

[0238] 2) Effects of Compound A27 on Gouty Arthritis Mice:

[0239] In this study, we investigated the manifestation of urate (MSU)-induced gouty arthritis model in Balb / c male mice (18-20 g) housed in a specific pathogen-free clean environment. The mice were randomly assigned to three different experimental groups and received the following treatments: (1) The control group received an intra-articular injection of 20 μL of sterile saline into the right ankle joint; (2) The model group received an intraperitoneal injection of 100 μL of ATP solution (10 mM) and 30 minutes later received an intra-articular injection of 20 μL of MSU crystal suspension (25 mg / ml) into the right ankle joint; (3) The drug group received oral administration of A27 (10 mg / kg) 1 hour before the injection of ATP and MSU. Twelve hours after administration, the inflammatory response of the mice was assessed by measuring the ankle circumference and calculating the joint swelling index. The joint swelling index was calculated as follows: (post-treatment circumference - initial circumference) / initial circumference. In addition, we extracted samples from mouse serum and measured the levels of inflammatory factors IL-1β and TNF-α by ELISA experiments.

[0240] As shown in Figure 6, after 12 hours, the joint swelling of the model group mice was significantly higher than that of the control group, while the joint swelling of the mice in the A27 treatment group was significantly alleviated (see Figures AB). By analyzing the levels of IL-1β (see Figure C) and TNF-α (see Figure D) in the mouse serum, we found that the levels of inflammatory cytokines in the model group were significantly higher than those in the control group, while the release of inflammatory factors in the A27 treatment group was significantly suppressed.

[0241] In summary, the present application provides a P2X7 receptor antagonist shown in Formula I. The compound provided herein not only has excellent antagonistic activity against the P2X7 receptor, but also has excellent metabolic stability and pharmacokinetic properties. Compared with the positive control JNJ47965567, the metabolic stability and pharmacokinetic properties of the compound of the present application are significantly improved, and the drugability is significantly better. The compound provided herein can be used to prepare P2X7 receptor antagonists, as well as to prepare drugs for treating inflammation and inflammation-related diseases, kidney disease, gout, respiratory diseases, cancer, pain, central nervous system diseases, radiation brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis, allergic asthma and other diseases, and has broad application prospects.

Claims

1. The compound represented by formula I, its salt, its isotope compound, its stereoisomer or its hydrate: in, n is 0, 1, 2 or 3; R1 is selected from one or more R 1a Substituted from the following groups: 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 8-membered saturated cycloalkyl, 3- to 8-membered saturated heterocyclic group; R 1a are each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, nitro, carboxyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, SO2R 1b ; R 1b Selected from C 1～8 alkyl; Y is CH; X is N, R2 is nothing; R3 is selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 10-membered aryl or 5- to 10-membered heteroaryl; R4 is selected from the following groups substituted by one or more substituents: 5-6 membered aryl, 5-6 membered heteroaryl, fused heterocyclic group; the substituents are each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, SH, j R h Substituted with the following groups: SR k , 5- to 6-membered aryl, 5- to 6-membered heteroaryl, 5- to 6-membered saturated cycloalkyl, 5- to 6-membered saturated heterocyclic group, R k is selected from 5-6 membered aryl, 5-6 membered heteroaryl, j is 0, 1, 2, or 3; R h are independently selected from hydrogen, halogen, hydroxyl, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, cyano, amino, NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl, 5-6 membered heteroaryl, R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

2. The compound of formula I according to claim 1, its salt, its isotope compound, its stereoisomer or its hydrate, wherein R1 is selected from 1a Substituted with the following groups: And / or, R4 is selected from the following groups substituted by one or more of the substituents:

3. The compound of formula I according to claim 1, its salt, its isotope compound, its stereoisomer or its hydrate, wherein the structural formula of the compound is as shown in formula II: in, n is 0, 1 or 2; Y6 is selected from N, CR s6 ; Y7 is selected from N, CR s7 ; Y8 is selected from N, CR s8 ; Y9 is selected from N, CR s9 ; Y 10 Selected from N, CR s10 ; R s6 , R s7 , R s8 , R s9 , R s10 are each independently selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, carboxyl, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy, 3- to 8-membered cycloalkyl.

4. The compound of formula I according to claim 1, its salt, its isotope compound, its stereoisomer or its hydrate, wherein the structure of the compound is as shown in formula II-1 or formula II-2 or formula II-3: in, n is 0 or 1; Y6 is selected from N, CR s6 ; Y7 is selected from N, CR s7 ; Y8 is selected from N, CR s8 ; Y9 is selected from N, CR s9 ; Y 10 Selected from N, CR s10 ; R s6 , R s7 , R s8 , R s9 , R s10 are each independently selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, carboxyl, C 1～5 Alkyl, halogenated C 1～5 Alkyl, C 1～5 Alkoxy, halogenated C 1～5 Alkoxy, 3- to 8-membered cycloalkyl; R3 is selected from hydrogen, C 1～5 Alkyl, 3-5 membered cycloalkyl; R 4a and R 4b are each independently selected from hydrogen, cyano, halogen, hydroxyl, amino, C 1～8 Alkyl, 3-8 membered cycloalkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, with j R h Substituted groups: 5-6 membered aryl, 5-6 membered heteroaryl, 5-6 membered saturated cycloalkyl, 5-6 membered saturated heterocyclic group, j is 0, 1, 2, or 3; R h are independently selected from hydrogen, halogen, hydroxyl, C 1～8 Alkyl, halogenated C 1～8 Alkyl, C 1～8 Alkoxy, halogenated C 1～8 Alkoxy, cyano, amino, NHR 4c NR 4c R 4c NHSO2R 4c 、SO2R 4c 、MSO2R 4c 、MNHSO2R 4c ; M is selected from 5-6 membered aryl and 5-6 membered heteroaryl; R 4c Selected from C 1～8 Alkyl, 3- to 8-membered saturated cycloalkyl, and 3- to 8-membered saturated heterocyclic group.

5. The compound of formula (I) according to claim 4, its salt, its isotope compound, its stereoisomer or its hydrate, R 4a Selected from hydrogen, R 4b Selected from hydrogen, 6. The compound of formula I according to claim 5, its salt, its isotope compound, its stereoisomer or its hydrate, wherein the structure of the compound is as shown in formula II-4 or formula II-5: in, R3 is selected from hydrogen, C 1～3 alkyl; R S Selected from hydrogen, halogen, C 1～3 Alkyl, halogenated C 1～3 Alkyl, C 1～3 Alkoxy, halogenated C 1～3 Alkoxy, 3- to 5-membered saturated cycloalkyl.

7. The compound of formula I according to claim 6, its salt, its isotope compound, its stereoisomer or its hydrate, wherein the structure of the compound is as shown in formula II-6, formula II-7, formula II-8 or formula II-9:

8. The compound of formula I according to claim 1, its salt, its isotope compound, its stereoisomer or its hydrate, wherein the compound is one of the following compounds:

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8, its salt, its isotope compound, its stereoisomer or its hydrate as an active ingredient, and a pharmaceutically acceptable excipient.

10. Use of the compound according to any one of claims 1 to 9, its salt, its isotope compound, its stereoisomer or its hydrate in the preparation of a P2X7 receptor antagonist.

11. The use according to claim 10, wherein the P2X7 receptor antagonist is a drug for treating inflammation and inflammation-related diseases, gout, kidney disease, respiratory diseases, cancer, pain, central nervous system diseases, radiation brain damage, cerebral ischemia, myocardial damage, diabetes, depression, lupus erythematosus, atherosclerosis or allergic asthma.

12. The use according to claim 10, wherein the inflammation and inflammation-related diseases are preferably neurological inflammation, arthritis, colitis, pancreatitis fibrosis, alcoholic fatty hepatitis, bronchitis, pneumonia, lumbar spondylitis, vasculitis; The respiratory disease is an airway obstructive disease; The gout is acute gouty arthritis, intermittent gout, chronic gout, hyperuricemia, secondary gout, gouty nephropathy, and urinary tract stones; The kidney disease is primary glomerular disease, secondary glomerular disease, interstitial nephritis, renal tubular disease, renal vascular disease, hereditary kidney disease, acute kidney injury caused by ischemia-reperfusion, acute kidney injury caused by sepsis, drug-induced acute kidney injury, contrast agent-induced acute kidney injury, and chronic renal failure; The cancer is prostate cancer, breast cancer, lung cancer, ovarian cancer, pancreatic cancer, intestinal cancer, colon cancer, stomach cancer, skin cancer, brain tumor, leukemia, lymphoma; The pain is headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone pain, pain caused by cancer and tumor invasion, and neuropathic pain syndrome; The central nervous system diseases are Alzheimer's disease, Parkinson's syndrome, epilepsy, multiple sclerosis and other demyelinating syndromes, cerebral atherosclerosis, myasthenia gravis; The lupus erythematosus is systemic lupus erythematosus.

13. The use according to claim 11, wherein the airway obstructive disease is bronchial asthma, allergic asthma, intrinsic asthma, exogenous asthma, exercise-induced asthma, and drug-induced asthma; The primary glomerular disease is preferably focal segmental glomerulosclerosis, crescentic nephritis, minimal change nephropathy, IgA nephropathy, membranous nephropathy; The secondary glomerular disease is preferably ANCA-associated vasculitis, hypertensive nephropathy, diabetic nephropathy, hepatitis B-associated nephritis, lupus nephritis, purpura nephritis, hyperuricemia nephropathy, and lipoprotein nephropathy; The hereditary kidney disease is preferably polycystic kidney disease; The drug-induced acute kidney injury is preferably cisplatin-induced acute kidney injury, folic acid-induced acute kidney injury, and aristolochic acid-induced acute kidney injury.

14. Use of the compound according to any one of claims 1 to 9, its salt, its isotope compound, its stereoisomer or its hydrate in treating diseases related to P2X7 receptor.

15. Use of the compound according to any one of claims 1 to 9, its salt, its isotope compound, its stereoisomer or its hydrate in a method for treating a disease associated with P2X7 receptor.

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