Programmed cell necrosis inhibitor as well as preparation method and application thereof

Abstract

The invention provides a programmed cell necrosis inhibitor as well as a preparation method and application thereof. Specifically, the invention provides a compound, or pharmaceutically acceptable salts, hydrates or solvates thereof, and the compound is as shown in a formula I in the specification. The invention also provides a preparation method of the compound and a pharmaceutical composition containing the compound or application of the compound in treatment or prevention of programmed necrosis of cells and / or human receptor interacting protein 1 kinase (RIPK1) related diseases or symptoms. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of small molecule compounds, and in particular relates to a class of inhibitors of programmed cell necrosis and a preparation method and use thereof. Background Art

[0002] In the process of development and aging, the human body is always accompanied by dynamic regulation of cell proliferation and death. Active cell death is indispensable in physiological activities such as normal development, resistance to invasion of pathogenic microorganisms, and maintenance of internal environment homeostasis. Its disorder often leads to developmental malformations, immune system diseases, neurodegenerative diseases, cancer and other diseases, and even individual death. Therefore, intervention in programmed cell death is of great significance for disease treatment research. Apoptosis is the first mechanism of programmed cell death to be elucidated. In recent years, programmed cell necrosis has become a new hotspot in the field of cell death. Many studies have reported that programmed cell necrosis, an important pathological feature, is accompanied by a variety of degenerative diseases (such as Alzheimer's disease (AD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), retinal degenerative diseases, etc.), ischemia-reperfusion injury (cerebral infarction, myocardial infarction, etc.) and pathogen infection.

[0003] On the other hand, the activation of programmed cell necrosis pathways is often accompanied by inflammation. Persistent inflammation may cause irreversible damage to human tissues or organs, and also promote the progression of cardiovascular disease, diabetes, cancer, non-alcoholic fatty liver disease, degenerative diseases or metabolic syndrome, and is described as the root of all diseases. Various chronic inflammatory diseases, such as rheumatoid arthritis (RA), inflammatory bowel disease (IBD), psoriasis or hepatitis, not only seriously affect the health and quality of life of patients, but may also trigger fatal myocardial infarction or cancer; and acute inflammatory storms caused by infection, such as sepsis or severe pneumonia, may directly endanger life; tumor immunotherapy using CAR-T cells or PD-1 / PD-L1 antibodies often has varying degrees of acute inflammatory side effects, which seriously affect the quality of life or life safety of patients. In addition, programmed cell necrosis and inflammation are also involved in the regulation of the tumor microenvironment: lung cancer cells can induce programmed necrosis in specific cells of the vascular wall so that they can pass through the circulatory system and metastasize; the main components of necrosomes are highly expressed in pancreatic cancer, which can induce the expression of chemokine CXCL1 and inhibit the body's immune response. Therefore, inhibiting the occurrence of programmed cell necrosis is recognized to help treat and alleviate a variety of diseases.

[0004] Studies have shown that tumor necrosis factor α (TNF-α) is one of the main ways to stimulate programmed necrosis and inflammation in cells in the body, and its downstream signaling pathway is also the necrosis signaling pathway with the most clear mechanism. In this pathway, RIPK1 plays a key regulatory role: by forming signal complex I (Complex I) with TNFα receptor protein TNFR1 and other regulatory factors (TRADD, TRAF2, cIAP1 / 2, etc.), it activates the IKK complex and NF-κB inflammatory pathway; after kinase activity is activated, it enters the cytoplasm and forms Complex IIa with apoptotic factors (FADD, Caspase-3, etc.) and mediates the occurrence of cell apoptosis; or it activates RIPK3 and polymerizes with it to form necrotic bodies Complex IIb, ultimately inducing the occurrence of programmed cell necrosis (Necroptosis). During the process of programmed necrosis, the cell membrane structure will be destroyed and a large number of inflammatory factors will be released, thereby inducing a chain reaction of inflammation and cell death. Studies have shown that genetic mutations in the RIPK1 gene (D324V, D324H) cause abnormal accumulation and activation of RIPK1 protein, leading to the occurrence of autoimmune diseases; during the aging process, mutation and inactivation of upstream inhibitory regulatory factors (such as OPTN, TBK1, TAK1, A20, CYLD, etc.) can also lead to RIPK1 activation, thereby aggravating degenerative diseases and inflammatory conditions; in a variety of fungal or viral infections and organ ischemic reperfusion injuries, RIPK1 is also abnormally activated and induces inflammatory cell death. In the treatment of central nervous system diseases, antibody drugs have the disadvantages of being difficult to penetrate the blood-brain barrier and immunogenicity, while many existing targeted small molecule drugs may affect the normal function of neuronal cells; at the same time, the inhibition of certain key immune pathways will inevitably affect the normal immune function of the human body, thereby bringing the risk of serious infection. Therefore, RIPK1 kinase is recognized as a potential therapeutic target for a variety of degenerative or inflammatory diseases.

[0005] In preclinical studies, the first-in-class RIPK1 inhibitor Necrostatin-1 (Nec-1) and its analogs have demonstrated clear therapeutic effects on a variety of degenerative diseases, inflammation, cancer and other diseases. For example, it has a relieving effect on AD, ALS, MS, Parkinson's disease (PD), Huntington's disease (PD), inflammatory bowel disease, age-related macular degeneration, etc.; it has a protective effect on psoriasis, retinitis pigmentosa, inflammatory bowel disease, autoimmune diseases, bombesin-induced acute pancreatitis and sepsis / systemic inflammatory response syndrome (SIRS); it can effectively relieve ischemic brain injury, ischemic myocardial injury, retinal ischemia / reperfusion injury, retinal detachment-induced photoreceptor cell necrosis, glaucoma, renal ischemia-reperfusion injury, cisplatin-induced renal injury and traumatic brain injury; it can at least partially relieve other diseases associated with RIPK1-dependent apoptosis, necrosis or cytokine production, including blood and solid organ malignancies, bacterial infections and viral infections (including tuberculosis, influenza, etc.) and lysosomal storage diseases (especially Gaucher disease). Currently, multiple RIPK1 inhibitors are also in clinical trials for the treatment of a variety of autodegenerative or immune diseases. For example, Sanofi's SAR443820 is in Phase II clinical trials for ALS and MS, and its SAR443122 is in Phase II clinical trials for lupus erythematosus and ulcerative bowel disease.

[0006] However, existing RIPK1 inhibitors all have deficiencies to varying degrees. On the one hand, chronic diseases related to programmed cell necrosis, especially degenerative diseases related to the central nervous system, have high requirements for drug safety. For example, SAR443060, which was originally in Phase I clinical trials by Sanofi, was still found to have chronic toxic side effects in primates after reaching the endpoint of Phase I, and had to be replaced by SAR443820 for clinical trials. On the other hand, most RIPK1 inhibitors in clinical trials are analogs of GSK2982772, and their shared structural skeleton limits the optimization space for drugability. Such inhibitors often cannot effectively inhibit RIPK1 in non-primates, which also limits their verification in preclinical disease models, and is not conducive to the selection of clinical indications. Finally, RIPK1 inhibitors with both high activity and brain permeability are very limited, and their long-term drug safety also needs to be further verified. Therefore, small molecule RIPK1 kinase activity inhibitors that are different from the chemical skeleton of GSK2982772 and have both high activity, high specificity and high blood-brain barrier penetration are still the difficulties and hot spots in the creation of new drugs for the clinical treatment of necroptosis-related diseases. In summary, the field urgently needs new RIPK1 inhibitors and / or necroptosis inhibitors with novel chemical structures and more prominent pharmacokinetic and pharmacodynamic properties as candidate drugs for the prevention and treatment of diseases involving cell death and / or inflammation. Summary of the invention

[0007] The purpose of the present invention is to provide a class of necroptosis inhibitors as candidate drugs for preventing and treating diseases or disorders associated with necroptosis and / or human receptor interacting protein 1 kinase (RIPK1).

[0008] In a first aspect of the present invention, there is provided a compound as shown in the following formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof:

[0009]

[0010] Where:

[0011] X1 is selected from the group consisting of CH, N or a chemical bond;

[0012] X2, X3, X4 and X5 are each independently selected from the group consisting of CH, N;

[0013] Provided that the ring formed by X1, X2, X3, X4 and X5 is an aromatic ring;

[0014] M is selected from the group consisting of O, S, NR3, CHR3 or C(R3)2;

[0015] W and U are each independently selected from the group consisting of O, S, NR4, CHR4 or C(R4)2;

[0016] The A ring and the B ring are each independently selected from the following group: a substituted or unsubstituted C6-C10 aryl group, a substituted or unsubstituted 5-12 membered heteroaryl group;

[0017] R1 and R2 are each independently selected from the group consisting of none, H, substituted or unsubstituted C1-C6 alkyl, halogen;

[0018] When M is NR3, CHR3 or C(R3)2, R2 and R3 may form a substituted or unsubstituted 5-7 membered ring together with the C atom or N atom to which they are connected and -CC(O)-;

[0019] R3 and R4 are selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, halogen;

[0020] R6 is selected from the following group: H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 olefin, substituted or unsubstituted C2-C6 alkyne, or -CH≡CR5; wherein R5 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-12 membered heterocyclyl;

[0021] R7 is selected from the following group: H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 olefin, substituted or unsubstituted C2-C6 alkyne, or -CH≡CR8; wherein R8 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-12 membered heterocyclyl;

[0022] The substitution refers to that the hydrogen atoms on the group are replaced by one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, methylsulfone, -S(=O)2NH2, oxo (=O), -CN, hydroxyl, -NH 2. carboxyl, C2-C6 amide (-C(=O)-N(Rc)2 or -NH-C(=O)(Rc), Rc is H or C1-C5 alkyl), C1-C6 alkyl-(C2-C6 amide), C1-C6 amine, C6-C10 aryl, 5-7 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, 4-7 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O.

[0023] In another preferred embodiment, the formula I has a structure shown in the following formula II:

[0024]

[0025] M, W, U, A, B, R1, R6 and R7 are as defined above.

[0026] In another preferred embodiment, the A ring and the B ring are each independently selected from the following group: substituted or unsubstituted phenyl, substituted or unsubstituted 5-7 membered heteroaryl;

[0027] Substitution is as defined above.

[0028] In another preferred embodiment, the A ring and the B ring are each independently selected from the following group:

[0029]

[0030] In another preferred embodiment, the A ring is an unsubstituted phenyl group.

[0031] In another preferred embodiment, the B ring is an unsubstituted phenyl group.

[0032] In another preferred embodiment, the M is NR3, and / or

[0033] W is CHR4;

[0034] R6 is selected from the group consisting of H, halogen, CN, substituted or unsubstituted C1-C6 alkyl;

[0035] R7 is selected from the following group: H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkyne, or -CH≡CR8; wherein R8 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-12 membered heterocyclyl;

[0036] R3, R4 and substitution are as defined above.

[0037] In another preferred embodiment, R7 is selected from the following group: H, halogen, CN, -CH≡CR8; wherein R8 is selected from the following group: substituted or unsubstituted 5-12-membered nitrogen-containing heteroaryl, substituted or unsubstituted 4-8-membered nitrogen-containing heterocyclic group.

[0038] In another preferred embodiment, the formula I has a structure shown in the following formula III:

[0039]

[0040] M, W, U, A, B, X5, R1, R2, R3, R6 and R8 are defined as above.

[0041] In another preferred embodiment, the formula I has a structure shown in the following formula II:

[0042]

[0043] M, W, U, A, B, R1, R6 and R8 are as defined above.

[0044] In another preferred embodiment, the compound shown in formula I is selected from the following table:

[0045]

[0046]

[0047]

[0048]

[0049] The second aspect of the present invention provides a pharmaceutical composition, which comprises (a) a therapeutically effective amount of a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, hydrate or solvate thereof; and (b) a pharmaceutically acceptable carrier.

[0050] In the fourth aspect of the present invention, there is provided a use of the compound as described in the first aspect or the pharmaceutical composition as described in the third aspect in the preparation of a medicament for treating or preventing diseases or conditions associated with programmed cell death and / or human receptor interacting protein 1 kinase (RIPK1).

[0051] In another preferred embodiment, the compound or pharmaceutical composition treats or prevents the disease or condition by inhibiting human receptor interacting protein 1 kinase (RIPK1).

[0052] In another preferred embodiment, the human receptor interacting protein 1 kinase (RIPK1) includes RIPK1 in an inactive (or unactivated) state and RIPK1 in an activated state.

[0053] In another preferred embodiment, the compound or pharmaceutical composition can also treat or prevent (especially treat) the disease or condition by inhibiting the activated human receptor interacting protein 1 kinase (RIPK1).

[0054] In another preferred embodiment, the inhibition of human receptor interacting protein 1 kinase (RIPK1) includes one or more of the following: inhibiting the activity of RIPK1, or inhibiting the phosphorylation of RIPK1.

[0055] In another preferred embodiment, the compound or pharmaceutical composition treats or prevents the disease or condition by inhibiting the programmed necrosis pathway.

[0056] In another preferred embodiment, the inhibition of programmed necrosis pathway includes one or more of the following: inhibiting the activity of RIPK1, inhibiting the phosphorylation of RIPK1, or inhibiting the phosphorylation of MLKL.

[0057] In another preferred embodiment, the disease or condition is selected from one or more of the following groups: degenerative diseases, inflammation, ischemia-reperfusion injury, pathogen infection, Parkinson's disease (PD), age-related macular degeneration, autoimmune diseases, retinal detachment-induced photoreceptor cell necrosis, glaucoma, cisplatin-induced renal injury and traumatic brain injury, atherosclerosis caused by hyperlipidemia, other diseases related to RIPK1-dependent apoptosis, necrosis or cytokine production, bacterial infection, viral infection and lysosomal storage disease.

[0058] In another preferred embodiment, the degenerative diseases include: Alzheimer's disease (AD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), and retinal degenerative diseases.

[0059] In another preferred embodiment, the inflammation includes one or more of the following: enteritis, rheumatoid arthritis, psoriasis, retinitis pigmentosa, inflammatory bowel disease, inflammatory bowel disease (PD), bombesin-induced acute pancreatitis, sepsis / systemic inflammatory response syndrome (SIRS).

[0060] In another preferred embodiment, the ischemia-reperfusion injury includes one or more of the following: cerebral infarction, myocardial infarction, ischemic brain injury, ischemic myocardial injury, retinal ischemia / reperfusion injury, and renal ischemia-reperfusion injury.

[0061] In another preferred embodiment, the other diseases associated with RIPK1-dependent apoptosis, necrosis or cytokine production include one or more of the following: blood and solid organ malignancies.

[0062] In another preferred embodiment, the viral infection includes one or more of the following diseases or conditions: tuberculosis, influenza, coronavirus infection and pneumonia caused by it.

[0063] In another preferred embodiment, the lysosomal storage disease includes Gaucher disease.

[0064] In the fourth aspect of the present invention, a method for treating or preventing diseases or conditions associated with programmed cell death and / or human receptor interacting protein 1 kinase (RIPK1) is provided, the method comprising: administering a therapeutically effective amount of the compound as described in the first aspect or the pharmaceutical composition as described in the second aspect to a subject in need.

[0065] In the fifth aspect of the present invention, a method for inhibiting programmed cell necrosis is provided, comprising the steps of: culturing cells in the presence of the compound as described in the first aspect, thereby inhibiting programmed cell necrosis.

[0066] In another preferred embodiment, the method is non-therapeutic in vitro.

[0067] In the sixth aspect of the present invention, a method for inhibiting the activity of RIPK1 protein kinase is provided, comprising the step of contacting RIPK1 protein kinase with the compound according to the first aspect, thereby inhibiting the activity of RIPK1 protein kinase.

[0068] In another preferred embodiment, the method is non-therapeutic in vitro.

[0069] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Figure 1 The effect of the representative compound QY-18-26 on RIPK1 kinase activity is shown.

[0071] Figure 2 The pharmacokinetic properties of the representative compound QY-18-26 are shown.

[0072] Figure 3 The brain penetration properties of the representative compound QY-18-26 are shown.

[0073] Figure 4 The effects of the representative compound QY-18-26 on the systemic inflammatory response syndrome induced by TNFα are shown. DETAILED DESCRIPTION

[0074] After long and in-depth research, the inventors unexpectedly discovered a class of novel cell programmed necrosis inhibitors. The cell programmed necrosis inhibitor has excellent RIPK1 inhibitory activity. Therefore, it can be used to prepare a pharmaceutical composition for preventing and / or treating diseases involving cell death, RIPK1 and / or inflammation. In particular, the preferred compounds provided by the present invention also have excellent inhibitory activity on activated RIPK1, so compared with existing RIPK1 inhibitors that only inhibit inactivated RIPK1, the compounds provided by the present invention have faster improvement or treatment of diseases or conditions (such as inflammation) involving cell death and / or RIPK1 inflammation. Based on the above findings, the inventors have completed the present invention.

[0075] the term

[0076] Unless expressly stated otherwise, the terms used according to the present invention and herein have the following meanings:

[0077] The term "alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms. For example, a C1-C6 alkyl group refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, etc., or similar groups.

[0078] The term "cycloalkyl" refers to a cyclic alkyl group having a specified number of carbon atoms. For example, "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 1 to 8 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0079] The term "alkoxy" refers to an alkyl group as defined above, which is attached to the remainder of the molecule via an oxygen atom. For example, C1-C6 alkoxy refers to C1-C6 alkyl-O-.

[0080] The term "halogen" refers to F, Cl, Br and I.

[0081] The term "haloalkyl" refers to an alkyl group (alkyl group is as defined above) substituted with a halogen.

[0082] The term "aryl" refers to a polyunsaturated (usually aromatic) hydrocarbon group having a specified number of ring atoms, which may be a single ring or multiple rings (such as a bicyclic ring) fused together or linked covalently. Examples of aryl include, but are not limited to, phenyl and naphthyl.

[0083] The term "heteroaryl" refers to an aryl group (or ring) having a specified number of ring atoms and containing 1 to 5 (e.g., 1, 2, 3, 4, or 5) heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized; for example, a 5-10 membered heteroaryl group (or ring) refers to a heteroaryl group (or ring) containing 5, 6, 7, 8, 9, or 10 ring atoms. As used herein, a nitrogen-containing heteroaryl group refers to a heteroatom in which at least one of the heteroatoms is a nitrogen heteroatom. The heteroaryl group can be attached to the rest of the molecule via a heteroatom. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridine, benzothiazolyl, benzofuranyl, benzothiophenyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furanyl, thienyl.

[0084] The term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic response), ie, at a reasonable benefit / risk ratio.

[0085] The term "effective amount" refers to the amount of a therapeutic agent that treats, alleviates or prevents a target disease or condition, or an amount that exhibits a detectable therapeutic or preventive effect. The exact effective amount for a subject depends on the subject's size and health, the nature and extent of the condition, and the therapeutic agent and / or combination of therapeutic agents selected for administration. Therefore, it is not useful to specify an exact effective amount in advance. However, for a given condition, the effective amount can be determined by routine experimentation, which is within the judgment of the clinician.

[0086] Unless otherwise specified, in the present invention, all compounds appearing are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). Among all compounds of the present invention, each chiral carbon atom can be optionally in R configuration or S configuration, or a mixture of R configuration and S configuration.

[0087] The term "compound of the present invention" refers to a compound of formula I. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of formula I.

[0088] The term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, benzenesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid.

[0089] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of the compound of the present invention with water.

[0090] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the unrecorded specific conditions in the following examples are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0091] Synthesis Example 1

[0092] Method 1:

[0093] Synthesis of compound QY-12-88:

[0094]

[0095] 1-Methyl-1H-1,2,3-triazole-4-carboxylic acid methyl ester (QY-12-88): 1,2,3-triazole-4-carboxylic acid methyl ester (1.27 g, 10.0 mmol) was dissolved in 20 mL N,N-dimethylformamide in a 100 mL round-bottom flask, potassium carbonate (0.83 g, 6.0 mmol) was added at room temperature, iodomethane (1.50 g, 10.5 mmol) was added dropwise to the reaction solution under ice bath conditions, stirred for 1 hour, and then gradually returned to room temperature and continued to stir for 16 hours. The reaction was stopped when the substrate was completely consumed by LC-MS real-time monitoring. After the organic solvent was removed by rotary evaporation, the reaction solution was diluted with dichloromethane and transferred to a separatory funnel, washed with distilled water (10 mL × 3), extracted with dichloromethane (20 mL × 3), the organic phases were combined, washed with saturated sodium chloride (10 mL × 2), and dried with anhydrous sodium sulfate. The organic phase was filtered and rotary evaporated to remove dichloromethane, and separated and purified by silica gel column chromatography (PE: EA = 10–90%) to obtain 305 mg of white powdery solid with a yield of 22%. ESI-MS m / z 142.1 (M+H) + .

[0096] Synthesis of compound QY-12-95:

[0097]

[0098] 5-Benzyl-1-methyl-1H-1,2,3-triazole-4-carboxylic acid methyl ester (QY-12-95): 5 mL toluene and QY-12-88 (305 mg, 2.16 mmol) were added to a 15 mL pressure bottle, and palladium acetate (24 mg, 0.11 mmol), triphenylphosphine (113 mg, 0.43 mmol), potassium carbonate (746 mg, 5.40 mmol), pivalic acid (66 mg, 0.65 mmol) were added, and benzyl chloride (274 mg, 2.16 mmol) was added dropwise. The mixture was heated to 100 °C under nitrogen protection and reacted overnight. After the reaction raw materials were consumed, toluene was removed by rotary evaporation, and the reaction solution was diluted with ethyl acetate. After diatomaceous earth filtration, the organic solvent was removed by rotary evaporation, and silica gel column chromatography was used for separation and purification (PE: EA = 0-80%) to obtain 173 mg of light yellow transparent oily liquid with a yield of 35%. ESI-MS m / z 232.1(M+H) + .

[0099] Synthesis of compound QY-13-15:

[0100]

[0101] 5-Benzyl-1-methyl-1H-1,2,3-triazole-4-carboxylic acid (QY-13-15): QY-12-95 (173 mg, 0.75 mmol) was dissolved in 3 mL of tetrahydrofuran and transferred to an 8 mL pressure bottle. Lithium hydroxide monohydrate (63 mg, 1.50 mmol) was dissolved in 0.8 mL of distilled water and added dropwise to the reaction solution at room temperature. LC-MS monitored the reaction progress in real time. After 8 h, the raw material was consumed. All solvents in the reaction system were removed by rotary evaporation. The crude product can be directly used for subsequent synthesis without purification. ESI-MS m / z218.0 (M+H) + .

[0102] Synthesis of compound QY-13-17:

[0103]

[0104] (S)-5-Benzyl-N-(1-(4-iodophenyl)ethyl)-1-methyl-1H-1,2,3-triazole-4-carboxamide (QY-13-17): QY-13-15 (0.75 mmol) was dissolved in 5 mL of N,N-dimethylformamide in a 15 mL pressure bottle, HATU (341 mg, 0.90 mmol) and 4-iodobenzylamine (174 mg, 0.75 mmol) were added, and N,N-diisopropylethylamine (289 mg, 2.24 mmol) was added dropwise at room temperature, and stirring was continued at room temperature. LC-MS real-time monitoring, after the reaction is completed, add 15mL of dichloromethane to dilute the reaction solution, transfer to a separatory funnel, wash with 10mL of distilled water, extract with dichloromethane (10mL×3), combine the organic phases, wash with saturated sodium chloride (10mL×2), post-process the organic phase, dry it over anhydrous sodium sulfate, filter and spin dry to remove dichloromethane, separate and purify by silica gel column chromatography (PE:EA=0–100%) to obtain 153mg of white foam solid, with a yield of 47%. ESI-MS m / z 447.0(M+H) + .

[0105] Synthesis of compound QY-18-26:

[0106]

[0107] (S)-5-Benzyl-1-methyl-N-(1-(4-(pyridin-2-ylethynyl)phenyl)ethyl)-1H-1,2,3-triazole-4-carboxamide (QY-18-26): 1 mL of tetrahydrofuran and QY-13-15 (32 mg, 0.072 mmol) were added to an 8 mL pressure bottle in sequence. After stirring and dissolving evenly, 2-ethynylpyridine (11 mg, 0.11 mmol), cuprous iodide (2.7 mg, 0.014 mmol), N,N-diisopropylethylamine (37 mg, 0.288 mmol), and bis(triphenylphosphine)palladium dichloride (5 mg, 0.007 mmol) were added. The reaction was stirred at room temperature under nitrogen protection. After the reactants were consumed, the organic solvent was removed by rotary evaporation after filtration with diatomaceous earth, and purified by silica gel column chromatography (PE:EA=10–80%) to obtain 10 mg of a yellow-brown oily liquid with a yield of 33%. ESI-MS m / z 422.2 (M+H) + .

[0108] Method 2:

[0109] Synthesis of compound QY-16-97:

[0110]

[0111] 3-(tert-Butoxymethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (QY-16-97): 4-(tert-Butoxy)-3-oxobutanoic acid ethyl ester (2.98 g, 14.7 mmol) and dimethoxy-N,N-dimethylmethanamine (2.45 g, 20.6 mmol) were uniformly dissolved in 25 mL of toluene, heated to 65 ° C and stirred for reaction overnight, and then directly rotary evaporated to remove the organic solvent; 9 mL of glacial acetic acid was added to dissolve the reaction system, and hydrazine hydrate (1.03 g, 20.6 mmol) was slowly added dropwise to the reaction solution under stirring at room temperature, and the reaction was stirred at room temperature overnight. Add 250mL of ethyl acetate to dilute the reaction system, transfer to a separatory funnel, wash with saturated sodium bicarbonate solution (50mL×2), wash with distilled water (10mL×2), wash with saturated sodium chloride (10mL×2), dry with anhydrous sodium sulfate, filter and remove the organic solvent by rotary evaporation, and separate and purify by silica gel column chromatography (PE:EA=0–60%) to obtain 1.76g of orange-yellow oily liquid with a yield of 53%. ESI-MSm / z 227.1(M+H) + .

[0112] Synthesis of compound QY-16-98:

[0113]

[0114] 3-(tert-Butoxymethyl)-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (QY-16-98): QY-16-97 (1.76 g, 7.8 mmol) was dissolved in 35 mL of acetonitrile in a 100 mL round-bottom flask, cesium carbonate (6.34 g, 19.5 mmol) was added at room temperature, iodomethane (1.32 g, 9.3 mmol) was added dropwise to the reaction solution under ice bath conditions, and the mixture was gradually returned to room temperature and stirred for 10 h. LC-MS real-time monitoring showed that the substrate was completely consumed and the reaction was stopped. After the organic solvent was removed by rotary evaporation, the reaction solution was diluted with ethyl acetate and transferred to a separatory funnel, washed with distilled water (10 mL × 2), washed with saturated sodium chloride (10 mL × 2), and the organic phase was dried over anhydrous sodium sulfate, filtered and rotary evaporated to remove dichloromethane, and separated and purified by silica gel column chromatography (PE: EA = 0–80%) to obtain 624 mg of a light yellow transparent oily liquid with a yield of 33%. ESI-MS m / z 241.2 (M+H) + .

[0115] Synthesis of compound QY-17-11:

[0116]

[0117] 5-Benzyl-3-(tert-butoxymethyl)-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (QY-17-11): 10 mL toluene and QY-16-98 (603 mg, 2.51 mmol) were added to a 30 mL pressure bottle, and palladium acetate (56 mg, 0.25 mmol), triphenylphosphine (262 mg, 1.0 mmol), potassium carbonate (1214 mg, 8.79 mmol), pivalic acid (77 mg, 0.75 mmol) were added, and benzyl chloride (413 mg, 3.26 mmol) was added dropwise. The mixture was heated to 110 °C under nitrogen protection and reacted overnight. After the reaction raw materials were consumed, toluene was removed by rotary evaporation, and the reaction solution was diluted with ethyl acetate. After filtration with diatomaceous earth, the organic solvent was removed by rotary evaporation, and the mixture was separated and purified by silica gel column chromatography (PE: EA = 0-50%) to obtain 108 mg of light yellow transparent oily liquid with a yield of 13%. ESI-MS m / z331.1(M+H) + .

[0118] Synthesis of compound QY-17-54:

[0119]

[0120] 5-Benzyl-3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (QY-17-54): QY-17-11 (108 mg, 0.33 mmol) was directly dissolved in 2 mL of hydrogen chloride in dioxane solution, stirred at room temperature for 4 hours, and the reaction was complete. It was separated and purified by C18 reverse phase chromatography column (CH3CN:H2O=10-90%) to obtain 60 mg of light yellow transparent oily liquid, with a yield of 66%. ESI-MS m / z275.1 (M+H) + .

[0121] Synthesis of compound QY-17-69:

[0122]

[0123] 5-Benzyl-3-formyl-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (QY-17-69): QY-17-54 (86 mg, 0.31 mmol) was directly dissolved in 4 mL of dichloromethane, and manganese dioxide powder (136 mg, 1.57 mmol) was added and heated to reflux and stirred for overnight reaction. The reaction was monitored by LC-MS. The solid was removed by membrane filtration and dichloromethane was removed by rotary evaporation to obtain a light yellow oily liquid product, which could be directly used for the next step without separation and purification. ESI-MS m / z 273.1 (M+H) + .

[0124] Synthesis of compound QY-18-57:

[0125]

[0126] (S)-5-Benzyl-3-(((1-(4-iodophenyl)ethyl)amino)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (QY-18-57): QY-17-69 (0.169 mmol) was mixed into 3 mL of 1,2-dichloroethane, added to a 15 mL pressure bottle and stirred; (S)-1-(4-iodophenyl)ethane-1-amine hydrochloride (62 mg, 0.22 mmol), glacial acetic acid (3 drops), sodium triacetoxyborohydride (90 mg, 0.42 mmol) were added, stirred at room temperature overnight, and the reaction was stopped after LC-MS monitoring when the reaction was complete. The organic solvent was removed by rotary evaporation, the reaction system was dissolved in dimethyl sulfoxide and then filtered through a filter membrane, and separated and purified by C18 reverse phase chromatography column (CH3CN:H2O=10-90%) to obtain 73 mg of colorless transparent oily liquid with a yield of 86%. ESI-MS m / z 504.1(M+H) + .

[0127] Synthesis of compound QY-18-59:

[0128]

[0129] (S)-5-Benzyl-3-(((1-(4-iodophenyl)ethyl)amino)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid (QY-18-59): QY-18-57 (73 mg, 0.145 mmol) was dissolved in 2 mL of ethanol and 2 mL of tetrahydrofuran, and transferred to a 15 mL pressure bottle. Sodium hydroxide (58 mg, 1.45 mmol) was dissolved in 1.0 mL of distilled water and added dropwise to the reaction solution at room temperature. After heating to 40°C, the reaction was stirred overnight. The reaction was stopped after LC-MS monitoring. 2N hydrochloric acid was added to acidify the reaction system to weak acidity, and the reaction was filtered through a C 18 Reverse phase chromatography column separation and purification (CH3CN:H2O=0-80%) gave 46 mg of colorless transparent oily liquid with a yield of 67%. ESI-MS m / z 476.0 (M+H) + .

[0130] Synthesis of compound QY-18-62:

[0131]

[0132] (S)-3-benzyl-5-(1-(4-iodophenyl)ethyl)-2-methyl-5,6-dihydropyrrolo[3,4-c]py razol-4(2H)-one(QY-18-62): QY-18-59 (46 mg, 0.097 mmol) was dissolved in 2.5 mL N,N-dimethylformamide in a 15 mL pressure bottle, HATU (44 mg, 0.116 mmol) was added, and N,N-diisopropylethylamine (50 mg, 0.388 mmol) was added dropwise at room temperature, and stirring was continued at room temperature. LC-MS real-time monitoring, after the reaction was completed, trifluoroacetic acid was added to the reaction solution to adjust it to weak acidity, and then filtered through a filter membrane and purified by C 18 Reverse phase chromatography column separation and purification (CH3CN:H2O=10-80%) gave 27 mg of colorless transparent oily liquid, with a yield of 61%. ESI-MS m / z 457.9 (M+H) + .

[0133] Synthesis of compound QY-18-63:

[0134]

[0135] (S)-3-benzyl-2-methyl-5-(1-(4-(pyridin-2-ylethynyl)phenyl)ethyl)-5,6-dih ydropyrrolo[3,4-c]pyrazol-4(2H)-one(QY-18-63): In a 15 mL pressure bottle, add 1 mL of tetrahydrofuran and QY-18-62 (46 mg, 0.097 mmol) in sequence. After stirring and dissolving evenly, add 2-ethynylpyridine (9.1 mg, 0.089 mmol), cuprous iodide (2.2 mg, 0.012 mmol), N,N-diisopropylethylamine (50 mg, 0.388 mmol), and bis(triphenylphosphine)palladium dichloride (4.1 mg, 0.006 mmol). Stir the reaction at room temperature under nitrogen protection. After the reactants were consumed, the organic solvent was removed by diatomaceous earth filtration and rotary evaporation, and then separated and purified by silica gel column chromatography (PE: EA = 10-100%) to obtain 21 mg of a yellow-brown oily liquid with a yield of 82%. ESI-MS m / z 433.2 (M+H) + .

[0136] According to the above method, by replacing different synthetic substrates, the compounds shown in Table 1 were obtained:

[0137] Table 1

[0138]

[0139]

[0140]

[0141]

[0142] The NMR data of the compounds shown in Table 1 are as follows:

[0143] QY-17-24: 1 H NMR (400MHz, DMSO-d6) δ9.06 (d, J=8.4Hz, 1H), 7.65

[0144] (t,J=1.8Hz,1H),7.42(dt,J=7.9,1.8Hz,2H),7.32–7.25(m,3H),7.24–7.15(m,3H),5. 16(p,J=7.3Hz,1H),4.41(q,J=15.3Hz,2H),3.89(s,3H),1.49(d,J=7.1Hz,3H).ESI-MS m / z 399.0(M+H) + .

[0145] QY-17-25: 1 H NMR(400MHz,DMSO-d6)δ8.92(d,J=8.5Hz,1H),7.42

[0146] (d,J=7.1Hz,2H),7.37–7.09(m,8H),5.18(p,J=7.3Hz,1H),4.42(q,J=15.3Hz,2H),3.88(s,3H),1.50(d,J=7.1Hz,3H).ESI-MS m / z 321.1(M+H) + .

[0147] QY-18-15: 1 H NMR(400MHz,DMSO-d6)δ8.96(d,J=8.3Hz,1H),7.73

[0148] –7.61(m,2H),7.33–7.13(m,7H),5.12(p,J=7.3Hz,1H),4.40(q,J=15.3Hz,2H),3.88(s,3H),1.48(d,J=7.1Hz,3H).ESI-MS m / z 446.9(M+H) + .

[0149] QY-18-25: 1 H NMR(400MHz,DMSO-d6)δ9.02(d,J=8.4Hz,1H),8.75

[0150] (d,J=2.1Hz,1H),8.59(dd,J=4.9,1.7Hz,1H),7.98(dt,J=8.0,2.0Hz,1H),7.56(d,J=8.0Hz,2H),7.52–7.44(m,3H),7.28(t,J=7.3Hz,2H),7.24–7.15(m,3H),5.21(q,J=7.4 Hz,1H),4.42(q,J=15.3 Hz,2H),3.89(s,3H),1.52(d,J=7.1 Hz,3H).ESI-MSm / z 422.2(M+H) + .

[0151] QY-18-26: 1 H NMR(400 MHz,DMSO-d6)δ9.03(d,J=8.4 Hz,1H),8.61

[0152] (d,J=4.8 Hz,1H),7.86(td,J=7.8,1.9 Hz,1H),7.64(d,J=7.8 Hz,1H),7.58(d,J=7.9 Hz,2H),7.50(d,J=7.9 Hz,2H),7.42(dd,J=7.6,4.9 Hz,1H),7.28(t,J=7.3Hz,2H),7.24–7.15(m,3H),5.22(p,J=7.1 Hz,1H),4.42(q,J=15.3 Hz,2H),3.89(s,3H),1.52(d,J=7.0 Hz,3H).ESI-MS m / z 422.2(M+H) + .

[0153] QY-18-27: 1 H NMR(400 MHz,DMSO-d6)δ9.04(d,J=8.3 Hz,1H),8.69

[0154] (s,2H),7.65(d,J=5.1 Hz,2H),7.60(d,J=8.1 Hz,2H),7.52(d,J=8.0 Hz,2H),7.28(t,J=7.3 Hz,2H),7.24–7.16(m,3H),5.22(p,J=7.2 Hz,1H),4.42(q,J=15.3Hz,2H),3.89(s,3H),1.52(d,J=7.1 Hz,3H).ESI-MS m / z 422.2(M+H) + .

[0155] QY-18-28: 1 H NMR(400 MHz,DMSO-d6)δ9.01(d,J=8.4 Hz,1H),7.89

[0156] (d,J=8.6 Hz,4H),7.64(dd,J=8.4,1.8 Hz,1H),7.48(pd,J=7.1,1.7 Hz,2H),7.28–7.22(m,2H),7.22–7.14(m,3H),5.35(p,J=7.2 Hz,1H),4.52–4.21(m,2H),3.88(s,3H),1.60(d,J=7.0 Hz,3H).ESI-MS m / z 371.2(M+H) + .

[0157] QY-18-41: 1H NMR(400 MHz,DMSO-d6)δ9.12(d,J=8.2 Hz,1H),7.80

[0158] (d,J=8.0 Hz,2H),7.61(d,J=8.0 Hz,2H),7.30–7.24(m,2H),7.24–7.12(m,3H),5.22(p,J=7.3 Hz,1H),4.40(q,J=15.3 Hz,2H),3.88(s,3H),1.51(d,J=7.1 Hz,3H).ESI-MS m / z 346.1(M+H) + .

[0159] QY-18-42: 1 H NMR(400 MHz,DMSO-d6)δ8.95(d,J=8.4 Hz,1H),7.40

[0160] (d,J=8.0 Hz,2H),7.34(d,J=7.9 Hz,2H),7.27(d,J=7.2 Hz,2H),7.20(dd,J=14.8,7.1 Hz,3H),5.18(p,J=7.1 Hz,1H),4.41(q,J=15.3 Hz,2H),3.88(s,3H),1.49(d,J=7.0 Hz,3H),1.45(s,6H).ESI-MS m / z 403.2(M+H) + .

[0161] QY-18-43: 1 H NMR(400 MHz,DMSO-d6)δ8.97(d,J=8.3 Hz,1H),8.05

[0162] (s,1H),7.67(s,1H),7.43(s,4H),7.28(t,J=7.3 Hz,2H),7.24–7.16(m,3H),5.19(p,J=7.2 Hz,1H),4.42(q,J=15.3 Hz,2H),3.88(s,3H),3.85(s,3H),1.50(d,J=7.0 Hz,3H).ESI-MS m / z 425.2(M+H) + .

[0163] QY-18-44: 1 H NMR(400 MHz,DMSO-d6)δ8.95(d,J=8.4 Hz,1H),7.37

[0164] (q,J=8.4 Hz,4H),7.28(dd,J=7.9,6.3 Hz,2H),7.24–7.13(m,3H),5.17(p,J=6.9 Hz,1H),4.41(q,J=15.3 Hz,2H),3.88(s,3H),3.83–3.76(m,2H),3.48–3.40(m,2H),2.87(tt,J=9.0,4.1 Hz,1H),1.90–1.76(m,2H),1.64–1.54(m,2H),1.49(d,J=7.0 Hz,3H).ESI-MS m / z 429.2(M+H) + .

[0165] QY-18-47: 1 H NMR(400 MHz,DMSO-d6)δ8.93(d,J=8.6 Hz,1H),7.36(d,J=8.0Hz,2H),7.33–7.24(m,4H),7.24–7.15(m,3H),5.16(p,J=7.4 Hz,1H),4.41(q,J=15.3Hz,2H),3.88(s,3H),1.57–1.50(m,1H),1.49(s,3H),0.92–0.82(m,2H),0.75–0.66(m,2H).ESI-MS m / z 385.1(M+H) + .

[0166] QY-18-49: 1 H NMR(400 MHz,DMSO-d6)δ8.98(d,J=8.4 Hz,1H),7.54–7.37(m,4H),7.31–7.24(m,2H),7.24–7.14(m,3H),5.18(p,J=7.2 Hz,1H),4.41(q,J=15.3 Hz,2H),4.12(s,1H),3.88(s,3H),1.49(d,J=7.1 Hz,3H).ESI-MS m / z 345.2(M+H) + .

[0167] QY-18-63: 1H NMR(400 MHz,DMSO-d6)δ8.61(d,J=4.8 Hz,1H),7.86(td,J=7.7,1.8 Hz,1H),7.65(d,J=7.8 Hz,1H),7.60(d,J=8.0 Hz,2H),7.42(dd,J=7.6,5.0 Hz,1H),7.38(d,J=8.0 Hz,2H),7.32(d,J=5.8 Hz,4H),7.23(dt,J=5.8,2.8 Hz,1H),5.46(q,J=7.2 Hz,1H),4.40(d,J=16.3 Hz,1H),4.19(s,2H),4.00(d,J=16.3 Hz,1H),3.78(s,3H),1.61(d,J=7.2 Hz,3H).ESI-MS m / z 433.2(M+H) + .

[0168] QY-18-77: 1 H NMR(400 MHz,DMSO-d6)δ9.43(s,1H),9.01(d,J=8.4Hz,1H),8.95(s,1H),7.49(q,J=8.2 Hz,4H),7.28(t,J=7.3 Hz,2H),7.24–7.14(m,3H),5.21(p,J=7.3 Hz,1H),4.42(q,J=15.4 Hz,2H),3.89(s,3H),1.51(d,J=7.1 Hz,4H).ESI-MS m / z412.1(M+H) + .

[0169] QY-18-102: 1 H NMR(400 MHz,DMSO-d6)δ8.62(d,J=4.8 Hz,1H),7.87

[0170] (td,J=7.7,1.8 Hz,1H),7.64(dd,J=17.6,7.9 Hz,3H),7.43(d,J=7.9 Hz,3H),7.36–7.26(m,4H),7.25–7.18(m,1H),5.97(q,J=7.1 Hz,1H),4.39(d,J=4.9 Hz,2H),3.64(s,3H),3.50–3.42(m,1H),3.12–3.04(m,1H),2.80–2.62(m,2H),1.54(d,J=7.1Hz,3H).ESI-MS m / z 447.2(M+H) + .

[0171] QY-19-53: 1 H NMR(400 MHz,DMSO-d6)δ8.98(d,J=8.4 Hz,1H),7.43

[0172] –7.34(m,4H),7.28(t,J=7.3 Hz,2H),7.24–7.15(m,3H),5.17(p,J=7.3 Hz,1H),4.53–4.29(m,2H),3.88(s,3H),3.45–3.39(m,2H),3.26–3.18(m,2H),3.15–3.09(m,1H),3.04–3.00(m,1H),2.29–2.19(m,1H),1.96–1.86(m,1H),1.49(d,J=7.1 Hz,3H).ESI-MS m / z 414.2(M+H) + .

[0173] QY-19-54: 1 H NMR(400 MHz,DMSO-d6)δ9.00(d,J=8.4 Hz,1H),8.77

[0174] (s,1H),7.50–7.36(m,4H),7.28(dd,J=8.0,6.4 Hz,2H),7.24–7.12(m,3H),5.18(p,J=7.1 Hz,1H),4.41(q,J=15.3 Hz,2H),4.15(t,J=8.8 Hz,2H),4.01–3.90(m,3H),3.88(s,3H),1.49(d,J=7.0 Hz,3H).ESI-MS m / z 400.2(M+H) + .

[0175] QY-19-55: 1 H NMR(400 MHz,DMSO-d6)δ8.98(d,J=8.4 Hz,1H),7.39

[0176] (q,J=8.4 Hz,4H),7.28(dd,J=8.0,6.4 Hz,2H),7.24–7.13(m,3H),5.17(p,J=7.2 Hz,1H),4.50–4.29(m,2H),3.88(s,3H),3.46–3.38(m,2H),3.20–3.00(m,3H),2.69(s,3H),2.42–2.32(m,1H),2.06–1.96(m,1H),1.49(d,J=7.0Hz,3H).ESI-MS m / z 428.2(M+H) + .

[0177] QY-19-56: 1 H NMR(400 MHz,DMSO-d6)δ8.99(d,J=8.4 Hz,1H),7.48

[0178] –7.35(m,4H),7.28(t,J=7.3 Hz,2H),7.24–7.13(m,3H),5.18(p,J=7.2 Hz,1H),4.41(q,J=15.3 Hz,2H),4.10–4.00(m,2H),3.88(s,3H),3.82–3.66(m,3H),2.64(s,3H),1.49(d,J=7.0 Hz,3H).ESI-MS m / z 414.2(M+H) + .

[0179] QY-19-60: 1 H NMR(400 MHz,DMSO-d6)δ8.98(d,J=8.4 Hz,1H),7.39

[0180] (q,J=8.0 Hz,4H),7.28(t,J=7.4 Hz,2H),7.23–7.15(m,3H),5.18(p,J=7.2Hz,1H),4.41(q,J=15.3 Hz,2H),4.10(t,J=6.6 Hz,1H),3.88(s,3H),3.06–2.98(m,1H),2.92–2.84(m,1H),2.16–2.04(m,1H),1.90–1.67(m,3H),1.49(d,J=7.1 Hz,3H).ESI-MS m / z 414.2(M+H) + .

[0181] QY-19-62: 1H NMR(400 MHz,DMSO-d6)δ8.98(d,J=8.4 Hz,1H),7.45

[0182] –7.34(m,4H),7.28(dd,J=8.1,6.6 Hz,2H),7.24–7.13(m,3H),5.17(p,J=7.3Hz,1H),4.41(q,J=15.3 Hz,2H),3.88(s,3H),3.39(s,1H),2.80(s,1H),2.41(s,0H),2.37(s,3H),2.13(q,J=7.3 Hz,1H),1.81(ddt,J=24.7,18.6,8.0Hz,3H),1.49(d,J=7.1 Hz,3H).ESI-MS m / z 428.2(M+H) + .

[0183] SYL-30-52: 1 H NMR(400 MHz,DMSO-d6)δ9.08–8.98(m,1H),7.82(d,J=3.0 Hz,1H),7.59(d,J=7.6 Hz,1H),7.43(d,J=7.4 Hz,1H),7.28(dt,J=7.2,3.5 Hz,2H),7.24–7.10(m,4H),5.28–4.90(m,1H),4.56–4.27(m,2H),3.89(d,J=2.7 Hz,3H),1.48(dd,J=6.8,2.8 Hz,3H).ESI-MS m / z 447.1(M+H) + .

[0184] SYL-30-56: 1 H NMR(400 MHz,DMSO-d6)δ9.01(d,J=8.4 Hz,1H),7.47

[0185] (s,1H),7.40(d,J=7.7 Hz,1H),7.35–7.13(m,7H),5.15(t,J=7.6 Hz,1H),4.41(q,J=15.3 Hz,2H),3.88(s,3H),1.49(d,J=7.0 Hz,3H),1.45(s,6H).ESI-MS m / z 403.2(M+H) + .

[0186] SYL-30-58: 1H NMR(400 MHz,DMSO-d6)δ9.04(d,J=8.4 Hz,1H),8.78

[0187] (d,J=2.2 Hz,1H),8.61(dd,J=4.9,1.7 Hz,1H),8.02(dt,J=8.0,2.0 Hz,1H),7.67(d,J=1.9 Hz,1H),7.54–7.45(m,3H),7.41(t,J=7.6 Hz,1H),7.30–7.24(m,2H),7.22–7.13(m,3H),5.20(p,J=7.1 Hz,1H),4.42(q,J=15.3 Hz,2H),3.89(s,3H),1.53(d,J=7.1 Hz,3H).ESI-MS m / z 422.2(M+H) + .

[0188] SYL-30-62: 1 H NMR(400 MHz,DMSO-d6)δ9.03(d,J=8.4 Hz,1H),7.66

[0189] (s,1H),7.53(d,J=7.7 Hz,1H),7.48(d,J=7.7 Hz,1H),7.40(t,J=7.7 Hz,1H),7.27(t,J=7.4 Hz,2H),7.23–7.14(m,3H),5.18(p,J=7.3 Hz,1H),4.51–4.32(m,2H),3.89(s,3H),1.51(d,J=7.0 Hz,3H).ESI-MS m / z 345.1(M+H) + .

[0190] SYL-30-65: 1 H NMR(400 MHz,DMSO-d6)δ8.99(d,J=8.4 Hz,1H),8.52

[0191] (s,1H),8.39(s,1H),7.48(s,1H),7.42(d,J=7.6 Hz,1H),7.35–7.24(m,4H),7.23–7.15(m,3H),5.15(p,J=7.2 Hz,1H),4.50–4.26(m,2H),3.88(s,3H),3.29–3.21(m,2H),3.10–2.93(m,3H),2.08–1.98(m,2H),1.81–1.70(m,2H),1.49(d,J=7.0 Hz,3H).ESI-MS m / z 428.2(M+H) + .

[0192] QY-20-1: 1 H NMR(400 MHz,DMSO-d6)δ8.91(d,J=8.0 Hz,1H),8.71–

[0193] 8.53(m,1H),7.87(td,J=7.8,1.9 Hz,1H),7.66(d,J=7.8 Hz,1H),7.61–7.52(m,3H),7.42(td,J=7.9,5.7 Hz,3H),7.33–7.21(m,3H),7.10(dd,J=7.4,2.0Hz,2H),7.01(d,J=2.1 Hz,1H),5.68(s,2H),5.14(p,J=7.2 Hz,1H),1.46(d,J=7.1 Hz,3H).ESI-MS m / z 407.1(M+H) + .

[0194] QY-20-2: 1 H NMR(400 MHz,DMSO-d6)δ8.62(d,J=5.0 Hz,1H),8.44(d,

[0195] J=7.9 Hz,1H),8.07(s,1H),7.88(td,J=7.7,2.0 Hz,1H),7.66(d,J=7.8 Hz,1H),7.59(dd,J=8.2,1.9 Hz,2H),7.51–7.38(m,3H),7.29–7.23(m,2H),7.21–7.11(m,3H),5.17(p,J=7.1 Hz,1H),4.46–4.35(m,2H),3.65(s,3H),1.47(d,J=7.0 Hz,3H).ESI-MS m / z 421.2(M+H) + .

[0196] QY-20-5: 1 H NMR(400 MHz,DMSO-d6)δ8.89(d,J=7.9 Hz,1H),8.70–

[0197] 8.56(m,1H),7.87(td,J=7.7,1.8 Hz,1H),7.66(dd,J=6.1,4.3 Hz,2H),7.56(d,J=7.9Hz,2H),7.48–7.35(m,3H),7.33–7.25(m,3H),7.09(dd,J=6.6,2.9Hz,2H),5.59–5.36(m,2H),5.12(p,J=7.2Hz,1H),1.42(d,J=7.0Hz,3H).ESI-MS m / z 425.2(M+H) + .

[0198] QY-20-18: 1 H NMR(400MHz,DMSO-d6)δ9.12(d,J=8.3Hz,1H),8.80

[0199] (d,J=5.7Hz,1H),8.66–8.56(m,1H),7.88(td,J=7.8,1.8Hz,1H),7.66(d,J=7.9Hz,1H),7.57(d,J=8.0Hz,2H),7.54–7.41(m,5H),7.34(t,J=7.4Hz,1H),7.29(d,J=7.9Hz,2H),7.17(d,J=5.7Hz,1H),5.12(p,J=7.2Hz,1H),1.48(d,J=7.0Hz,3H).ESI-MSm / z 420.1(M+H) + .

[0200] QY-20-53: 1 H NMR(400MHz,DMSO-d6)δ9.06(d,J=8.3Hz,1H),8.70

[0201] –8.53(m,1H),7.86(td,J=7.7,1.8Hz,1H),7.65(d,J=7.8Hz,1H),7.58(d,J=8.2Hz,2H),7.50(d,J=8.1Hz,2H),7.42(ddd,J=7.6,4.9,1.3Hz,1H),7.29–7.21(m,2H),7.16–7.07(m,2H),5.22(p,J=7.3Hz,1H),4.40(q,J=15.3Hz,2H),3.90(s,3H),1.52(d,J=7.1Hz,3H).ESI-MS m / z 440.2(M+H) + .

[0202] QY-20-54: 1 H NMR(400MHz,DMSO-d6)δ9.08(d,J=8.3Hz,1H),8.72

[0203] –8.55(m,1H),7.87(td,J=7.8,1.9Hz,1H),7.66(d,J=7.8Hz,1H),7.58(d,J=8.1Hz,2H),7.50(d,J=8.1Hz,2H),7.46–7.40(m,1H),7.33(q,J=7.4Hz,1H),7.11–7.00(m,3H),5.22(p,J=7.2Hz,1H),4.50–4.37(m,2H),3.92(s,3H),1.52(d,J=7.1Hz,3H).ESI-MS m / z 440.2(M+H) + .

[0204] QY-20-55: 1 H NMR(400MHz,DMSO-d6)δ9.06(d,J=8.3Hz,1H),8.90

[0205] (s,1H),7.96(d,J=1.4Hz,1H),7.60(d,J=8.1Hz,2H),7.52(d,J=8.1Hz,2H),7.28(dd,J=8.0,6.5Hz,2H),7.24–7.16(m,3H),5.21(p,J=7.2Hz,1H),4.51–4.33(m,2H),3.89(s,3H),3.86(s,3H),1.52(d,J=7.1Hz,3H).ESI-MS m / z 425.2(M+H) + .

[0206] Biological test example 1: Test of the inhibitory activity of RIPK1 inhibitor on programmed cell necrosis

[0207] The biological test scheme used was: the effect of the compound on TNF-induced FADD (Fas-associated death domain)-deficient Jurkat cells and L929 cells programmed necrosis.

[0208] In order to verify the inhibitory effect of the compounds of the present invention on programmed cell necrosis at the cellular level, cell types closely related to the RIP1 pathway, namely FADD-deficient Jurkat cells (human peripheral blood leukemia T cell line) and L929 cells, were selected, and two different stimulation methods were used: tumor necrosis factor (TNFα) alone, or TNFα combined with mitochondrial-derived cysteine ​​aspartate activator (SMAC) SM164. The cell viability was calculated by detecting the chemiluminescence value, thereby obtaining the biological activity of the compound in inhibiting programmed cell necrosis.

[0209] Methods: FADD-deficient Jurkat cells: FADD-deficient Jurkat cells (human peripheral blood leukemia T cell line) were cultured in vitro. After growing to the logarithmic growth phase, the cells were collected and centrifuged at 1000 rpm for 5 min. The supernatant was discarded and the cell concentration was adjusted to 2.5×10 5 / mL, the cells were seeded into 384-well plates, 40μl per well. SM164 (50nM) and 5μL of the compound diluted in cell culture medium were added to the corresponding wells. After pretreatment at 37°C for 1h, 5μL of TNFα (50ng / mL) diluted in cell culture medium was added to each well of the stimulation group, and 5μL of culture medium was added to the control group. After culturing in a cell culture incubator (37°C, 5% CO2) for 14h, 15μl of Cell Titer-Glo solution was added to each well, incubated at room temperature for 30min, and the chemiluminescence value (luminescence) was detected to measure the intracellular ATP level. The unstimulated DMSO control well was taken as 100% cell viability. L929 cells: L929 cells (mouse fibroblasts) cultured in vitro were digested and diluted to 6.25×10 4 / ml, cells were seeded into 384-well plates, 40μl per well. Place in a cell culture incubator (37°C, 5% CO2) and culture for 12h. Add 5μL of SM164 (500nM) and compound diluted with cell culture medium to the corresponding wells. After pretreatment at 37°C for 1h, 5μL of TNFα (500ng / mL) diluted with cell culture medium was added to each well of the stimulation group, and 5μL of culture medium was added to the control group. After culturing in a cell culture incubator (37°C, 5% CO2) for 14h, 15μl of CellTiter-Glo solution was added to each well, incubated at room temperature for 30min, and chemiluminescence was detected to measure the intracellular ATP level. The unstimulated DMSO control well was taken as 100% cell viability. Prism Graphpad statistical software was used to calculate the compound EC 50 The results are shown in Table 2.

[0210]

[0211] Table 2. Test results of the inhibitory activity of RIPK1 inhibitors on programmed cell necrosis

[0212]

[0213]

[0214] The experimental results in Table 2 show that: whether stimulated by TNFα alone or in combination with SM164, the preferred compounds of the present invention, such as QY-18-26, QY-18-27, QY-20-53 and QY-20-54, all exhibited stronger programmed necrosis inhibitory activity than the clinical inhibitor SAR443122 on human FADD-deficient Jurkat cells; and on mouse L929 cells where SAR443122 was almost inactivated, the preferred compounds of the present invention were still able to well inhibit the occurrence of programmed necrosis.

[0215] Biological test example 2: Testing the effect of the representative compound QY-18-26 on the kinase activity of RIPK1.

[0216] The biological test scheme used: The effect of the test compound QY-18-26 on the kinase activity of the RIPK1 (1-330) protein. The in vitro purified RIPK1 (1-330) protein retains the complete kinase activity domain and maintains good kinase activity. The known RIPK1 clinical inhibitor SAR443122 was used as a control.

[0217] Methods: RIPK1 (1-330) protein with a final concentration of 2 μM and ATP (1X kinase buffer) of the corresponding concentration were added to a 384-well plate, with a final concentration of 5 μL. At least 3 replicate wells were set up for each group, and the reaction was carried out at 37°C for 2 h. 5 μL ADP-Glo ​​reagent was added to stop the kinase reaction and remove the residual ATP in the reaction system. The reaction was allowed to stand at room temperature for 40 min. 10 μL Kinase Detection Reagent was added to convert ADP into ATP and introduce luciferase and luciferin for detecting ATP into the system. The reaction was allowed to react at room temperature for 1 h. Luminescence was detected using the 7500 Fast Real-Time PCR System. The IC of the compound inhibiting the kinase reaction was calculated using Prism Graphpad statistical software. 50 The experimental results can be found in Figure 1 .

[0218] The experimental results showed that the representative compound QY-18-26 exhibited effective concentration-dependent inhibition of RIPK1 kinase, with an effective half-inhibitory concentration of 6.4 nM, which was superior to the control SAR443122 (effective half-inhibitory concentration of 9.6 nM).

[0219] Biological Test Example 3: Pharmacokinetic Properties of Representative Compound QY-18-26

[0220] The biological test scheme used is: drug metabolism test of the compound in living mice.

[0221] In order to verify the pharmacokinetic properties of the compounds of the present invention in vivo, the representative compound QY-18-26 was selected and tested for its pharmacokinetic properties in mice (n=3) by single administration via oral gavage (PO, 10 mg / kg) or intravenous injection (IV, 1 mg / kg). The experimental results are shown in Figure 2 shown.

[0222] The results showed that after a single dose of QY-18-26, the peak concentration in mice was reached rapidly and a high maximum blood concentration (C max ), exposure (AUC) and bioavailability (F). During the 24 hours after oral administration of 10 mg / kg, the blood drug concentration could be stably maintained above the effective inhibitory concentration (5 ng / mL) for most of the time.

[0223] Biological Test Example 4: Brain Penetration Properties of Representative Compound QY-18-26

[0224] The biological testing scheme used is: brain / blood drug concentration testing of compounds in living mice and rats.

[0225] In order to verify the brain-penetrating property of the compounds of the present invention in vivo, the representative compound QY-18-26 was selected and the brain / blood drug concentration of the compound at different time points in mice (n=3) was tested by single administration of oral gavage (PO, 10 mg / kg) or intravenous injection (IV, 2 mg / kg); at the same time, the brain / blood drug concentration of the compound at different time points in rats (n=3) was tested by single administration of oral gavage (PO, 5 mg / kg). The experimental results are shown in Figure 2. Figure 3 shown.

[0226] After a single dose of QY-18-26, the concentration in the brain of mice and rats was approximately 40% of that in plasma, and it maintained a stable brain-blood distribution at 4 hours. Its brain drug concentration was far above the effective inhibitory concentration (5 ng / g), indicating that the compound has ideal brain penetrating properties.

[0227] Biological Test Example 5: Effect of Representative Compound QY-18-26 on Systemic Inflammatory Response Syndrome Induced by TNFα

[0228] Biological test scheme used: Systemic inflammatory response syndrome, also known as inflammatory storm, refers to a systemic nonspecific inflammatory response caused by infectious or non-infectious injuries such as severe infection, multiple trauma, burns, ischemia-reperfusion, acute pancreatitis, etc. Under this condition, a large number of inflammatory factors will be released, and in severe cases, the body will lose control of the inflammatory response, causing multiple organ failure and even death. Tail vein injection of tumor necrosis factor TNF can cause systemic inflammatory response syndrome in mice, resulting in a decrease in body temperature and death of mice. The representative compound QY-18-26 was tested for its effect on the systemic inflammatory response syndrome caused by TNFα, and the temperature changes and death of mice were monitored. The known RIPK1 inhibitor Nec-1s was used as a control, and the clinical inhibitor SAR443122 had poor inhibitory activity on mouse RIPK1, so it was not used.

[0229] Method: The compound to be tested was dissolved in 0.5% carboxymethyl cellulose according to the required concentration one day in advance and sonicated overnight. Each mouse was gavaged with 200 μl, and 20 minutes later, 10 mg TNFα (dissolved in 125 μl PBS) was injected through the tail vein. After injection, the temperature of the mice was monitored with an infrared thermometer. The effect of the compound was calculated using Prism Graphpad statistical software. The experimental results are shown in Figure 2. Figure 4 shown.

[0230] The experimental results showed that in the absence of inhibitors, all mice died within hours after being injected with TNFα; the compound QY-18-26 was able to effectively resist the hypothermia, inflammatory response and death caused by TNFα at a low dose (1 mg / kg).

Claims

1. A compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, characterized in that: The compound is shown in Formula I: Where: X1 is selected from the group consisting of CH, N or a chemical bond; X2, X3, X4 and X5 are each independently selected from the group consisting of CH, N; Provided that the ring formed by X1, X2, X3, X4 and X5 is an aromatic ring; M is selected from the group consisting of O, S, NR3, CHR3 or C(R3)2; W and U are each independently selected from the group consisting of O, S, NR4, CHR4 or C(R4)2; The A ring and the B ring are each independently selected from the following group: a substituted or unsubstituted C6-C10 aryl group, a substituted or unsubstituted 5-12 membered heteroaryl group; R1 and R2 are each independently selected from the group consisting of none, H, substituted or unsubstituted C1-C6 alkyl, halogen; When M is NR3, CHR3 or C(R3)2, R2 and R3 may form a substituted or unsubstituted 5-7 membered ring together with the C atom or N atom to which they are connected and -CC(O)-; R3 and R4 are selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, halogen; R6 is selected from the following group: H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 olefin, substituted or unsubstituted C2-C6 alkyne, or -CH≡CR5; wherein R5 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-12 membered heterocyclyl; R7 is selected from the following group: H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 olefin, substituted or unsubstituted C2-C6 alkyne, or -CH≡CR8; wherein R8 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-12 membered heterocyclyl; The substitution refers to that the hydrogen atoms on the group are replaced by one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, methylsulfone, -S(=O)2NH2, oxo (=O), -CN, hydroxyl, -NH 2. carboxyl, C2-C6 amide (-C(=O)-N(Rc)2 or -NH-C(=O)(Rc), Rc is H or C1-C5 alkyl), C1-C6 alkyl-(C2-C6 amide), C1-C6 amine, C6-C10 aryl, 5-7 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, 4-7 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, characterized in that: The compound is shown in Formula II: The definitions of M, W, U, A, B, R1, R6 and R7 are as described in claim 1.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, characterized in that: The A ring and the B ring are each independently selected from the following group: substituted or unsubstituted phenyl, substituted or unsubstituted 5-7 membered heteroaryl; Substitution is defined as in claim 1.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, characterized in that: The A ring and the B ring are each independently selected from the following group:

5. The compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, characterized in that: In the formula I, M is NR3, and / or W is CHR4; R6 is selected from the group consisting of H, halogen, CN, substituted or unsubstituted C1-C6 alkyl; R7 is selected from the following group: H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkyne, or -CH≡CR8; wherein R8 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-12 membered heterocyclyl; R3, R4 and substitution are as defined in claim 1.

6. The compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, characterized in that: The compound is shown in formula III: The definitions of M, W, U, A, B, X5, R1, R2, R3, R6 and R8 are as described in claim 1.

7. The compound according to claim 1, characterized in that The compound is selected from the following table:

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises (a) a therapeutically effective amount of the compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof; and (b) a pharmaceutically acceptable carrier.

9. Use of the compound according to claim 1 or the pharmaceutical composition according to claim 8 in the preparation of a medicament for treating or preventing a disease or condition associated with programmed cell death and / or human receptor interacting protein 1 kinase (RIPK1).

10. The use according to claim 9, characterized in that The disease or condition is selected from one or more of the following groups: degenerative diseases, inflammation, ischemia-reperfusion injury, pathogen infection, Parkinson's disease (PD), age-related macular degeneration, autoimmune diseases, retinal detachment-induced photoreceptor cell necrosis, glaucoma, cisplatin-induced renal injury and traumatic brain injury, atherosclerosis caused by hyperlipidemia, other diseases related to RIPK1-dependent apoptosis, necrosis or cytokine production, bacterial infection, viral infection and lysosomal storage disease.

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