Indazolone compound, preparation method therefor, pharmaceutical composition, and use

Abstract

Disclosed in the present invention are an indazolone compound, a preparation method therefor, a pharmaceutical composition, and a use. Specifically, provided are a compound of formula (I), or a pharmaceutically acceptable salt, tautomer, stereoisomer, isotopic compound, pharmaceutically usable salt, ester, prodrug or hydrate thereof, a preparation method therefor, a pharmaceutical composition, and a use. The compound of the present invention has good anti-tumor activity as a CRL4CRBNE3 ubiquitin ligase modulator, and can be used in the preparation of a drug for treating CRL4CRBN E3 ubiquitin ligase-related diseases.

Description

Indazolone compounds, preparation methods, pharmaceutical compositions and applications thereof Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a class of indazolone compound derivatives, preparation methods, pharmaceutical compositions and applications thereof. Background Art

[0002] Targeted protein degradation is a promising area for the discovery and development of innovative therapeutics. Targeted protein degradation technology transcends the mode of action of traditional small molecule inhibitors and can directly induce substrate proteins through the ubiquitin-proteasome protein degradation (UPS) pathway in vivo. Currently, targeted protein degradation technologies based on the ubiquitin-proteasome system (UPS) primarily include proteolysis targeting chimeras (PROTACs) and molecular glues.

[0003] Molecular glues are small molecules that induce protein-protein interactions and can exert precise temporal control over various biological processes. Molecular glues are small molecules that act on protein interfaces and form novel protein-protein interactions. Molecular glues mediate proximity-induced protein degradation and possess intrinsic advantages over heterobifunctional proteolysis-targeting chimeras, including unprecedented mechanisms, unique bioactivities, and favorable physicochemical properties. However, the paucity of serendipitously discovered molecular glues has hindered their development as reagents for bioassays and disease treatment. Therefore, rational discovery and design strategies are essential for the development of molecular glue-based drugs.

[0004] Currently, CRBN-based molecular glue degraders are mainly immunomodulatory drugs (IMiDs), such as thalidomide, lenalidomide, and pomalidomide. Because CRBN ligands and immunomodulatory imide drugs (IMiDs) have good drug-like properties, targeting CRBN is the main mode of action of molecular glue degraders. However, IMiDs are inherently unstable and easily hydrolyzed in body fluids. For example, thalidomide undergoes racemization and hydrolysis in body fluids, and the rate accelerates with increasing pH. Therefore, at pH 7, 7.4, and 8, the hydrolysis half-life of thalidomide is 11, 5, and 1.25 hours, respectively. Therefore, the development of small molecules with good pharmaceutical properties targeting CRBN is an important direction for the design of molecular glue degraders.

[0005] Lenalidomide is currently mainly used to treat multiple myeloma and myelodysplastic syndrome, but its efficacy in other indications is not very ideal; therefore, the development of a class of novel molecular glue degraders can, on the one hand, improve the therapeutic effect on tumors and expand new indications, which can be used to further address unmet clinical needs.CRBN New ligands and new molecular glues for E3 ubiquitin ligases have important research value and practical significance in the development of new drugs.

[0006] Summary of the Invention

[0007] The present inventors have found through literature research that indazolones and their derivatives have unique biological activities and good metabolic stability. CRBN Through molecular docking and computational simulation, we found that the new skeleton (3-(3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione) derived from the combination of indazolone and glutarimide ring can better bind to CRL4. CRBN The E3 ubiquitin ligase can bind to the pocket in the E3 ubiquitin ligase and reduce the steric repulsion with the carbonyl group on the nearby polypeptide chain in the binding protein, thus, it can serve as a kind of favorable binding to CRL4. CRBN Therefore, based on the good bioavailability and drug metabolism properties of indazolone structures, we designed a new skeleton by combining indazolone and glutarimide rings. This indazolone-derived glutarimide structure can bind well to CRL4. CRBN E3 ubiquitin ligase binding; various novel CRL4 structures can be obtained through structural modification CRBN E3 ubiquitin ligase molecular glue degrader.

[0008] The purpose of the present invention is to provide a novel indazolone skeleton structure CRL4 CRBN E3 ubiquitin ligase molecular glue degraders have improved tumor treatment effects and can broaden the indications of molecular glue degraders, and provide CRL4 with better physicochemical properties for the design and development of degraders of related pathogenic proteins. CRBN Ligand for E3 ubiquitin ligase.

[0009] One aspect of the present invention is to provide a compound represented by general formula (I), its enantiomers, diastereomers, racemates, isotopic compounds, metabolic precursors, metabolites, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof.

[0010] Another aspect of the present invention is to provide a method for preparing the compound represented by general formula (I), an important intermediate for preparing the compound and a preparation method thereof.

[0011] Another aspect of the present invention is to provide compounds as shown in the general formula (I) and their variants

[0012] Isomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof, characterized in that the compound is used to prepare a drug for preventing or treating CRL4 CRBN Drugs or diagnostic reagents for diseases related to E3 ubiquitin ligase; preferably, the CRBN Diseases associated with E3 ubiquitin ligases include cancer, pain, central nervous system diseases, and immune system diseases.

[0013] To achieve the above-mentioned object, the present invention provides a compound represented by the following general formula (I), or a pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof:

[0014] wherein R1 is hydrogen, methyl or ethyl;

[0015] R2 is hydrogen or fluorine;

[0016] R3 is hydrogen, deuterium or fluorine;

[0017] L is none, -CH2-, S, -CO-, -CH(CH3)-;

[0018] X-A1 combination is -(CH2) n -O-CH2-A1, -(CH2) n -NHCONH-A1, -(CH2) n -NHCOCF2-A1, -CONH-(CH2) n -A1, -CONH-CO-A1, -CONHCH(CH3)-A1, -CONHCH(CF3)-A1;

[0019] n is 0 or 1;

[0020] When the X-A1 combination is -(CH2) n -O-CH2-A1 or, L is none, -CH2-, S, -CO-, -CH(CH3)-;

[0021] A1 is selected from: halogen-substituted and unsubstituted phenyl, halogen-substituted and unsubstituted pyridyl, and five-membered heteroaryl containing N, O, and S;

[0022] A2 is selected from: phenyl, pyridyl, piperazinyl, piperidinyl, morpholinyl, azetidinyl; wherein the phenyl, pyridyl, piperazinyl, piperidinyl, morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, nitro, hydroxyl, carboxyl, aminocarbonyl, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy-substituted C1-C6 Alkoxy; wherein the substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy;

[0023] When the X-A1 combination is -(CH2) n -NHCONH-A1 or -(CH2) n -NHCOCF2-A1, L and A2 are both absent;

[0024] A1 is selected from: phenyl, wherein the phenyl is substituted by one or more groups selected from the following: hydrogen, halogen, cyano, C1-C6 alkyl;

[0025] When the X-A1 combination is -CONHCH(CH3)-A1, -CONH-(CH2) n When -A1, -CONH-CO-A1 or -CONHCH(CF3)-A1, L and A2 are not present at the same time;

[0026] A1 is selected from: phenyl, pyridyl, 5-10 membered heteroaryl, C3-C10 cycloalkyl, 4-6 membered heterocyclyl, 4-6 membered heterocyclylphenyl, C1-C6 alkyl; the aforementioned C6-10 aryl, 5-10 membered heteroaryl, C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl is substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy.

[0027] In another preferred embodiment, R1 is hydrogen, methyl or ethyl;

[0028] R2 is hydrogen or fluorine;

[0029] R3 is hydrogen, deuterium or fluorine;

[0030] L is none, -CH2-, S, -CO-, -CH(CH3)-;

[0031] X-A1 combination is -(CH2) n -O-CH2-A1, -(CH2) n -NHCONH-A1, -(CH2) n -NHCOCF2-A1, -(CH2) n -NHCH2-A1, -CONH-(CH2) n -A1, -CONHCH(CH3)-A1, -CONHCH(CF3)-A1, or X is absent;

[0032] n is 0 or 1;

[0033] When the X-A1 combination is -(CH2) n -O-CH2-A1 or, L is none, -CH2-, S, -CO-, -CH(CH3)-;

[0034] A1 is selected from: halogen-substituted and unsubstituted phenyl, halogen-substituted and unsubstituted pyridyl, and five-membered heteroaryl containing N, O, and S;

[0035] A2 is selected from: phenyl, pyridyl, piperazinyl, piperidinyl, morpholinyl; wherein the phenyl, pyridyl, piperazinyl, piperidinyl, morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, nitro, hydroxyl, carboxyl, aminocarbonyl, 4-10 membered heterocyclic radical, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl , C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy substituted C1-C6 alkoxy; wherein the substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy;

[0036] When the X-A1 combination is -(CH2) n -NHCONH-A1 or -(CH2) n -NHCOCF2-A1, L and A2 are both absent;

[0037] A1 is selected from: phenyl, wherein the phenyl is substituted by one or more groups selected from the following: hydrogen, halogen, cyano, C1-C6 alkyl;

[0038] When the X-A1 combination is -CONHCH(CH3)-A1, -CONH-(CH2)n When -A1 or -CONHCH(CF3)-A1, L and A2 are not present at the same time;

[0039] A1 is selected from: phenyl, pyridyl, 5-10 membered heteroaryl, C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl; the aforementioned C6-10 aryl, 5-10 membered heteroaryl, C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl is substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy.

[0040] In another preferred embodiment, the compound has a structure shown by formula (I-1)-(I-11):

[0041] wherein R1 is hydrogen, methyl or ethyl;

[0042] R2 is hydrogen or fluorine;

[0043] R3 is hydrogen, deuterium or fluorine;

[0044] R4 is H or halogen;

[0045] n = 0 or 1;

[0046] R5 is selected from phenyl, pyridyl or morpholinyl, wherein the phenyl, pyridyl or morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy;

[0047] A2 is selected from: phenyl, pyridyl; wherein the phenyl, pyridyl is optionally substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy substituted by C1-C6 alkoxy.

[0048] In another preferred embodiment, the compound has a structure shown by formula (I-29)-(I-30):

[0049] wherein R1 is hydrogen, methyl or ethyl;

[0050] R2 is hydrogen or fluorine;

[0051] R3 is hydrogen, deuterium or fluorine;

[0052] R 11 is hydrogen or methyl;

[0053] Each q is independently 0 or 1;

[0054] Z is N or -CH-;

[0055] R5 is selected from phenyl, pyridyl or morpholinyl, wherein the phenyl, pyridyl or morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy;

[0056] A2 is selected from: phenyl, pyridyl; wherein the phenyl, pyridyl is optionally substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy substituted by C1-C6 alkoxy.

[0057] In another preferred embodiment, R1 is hydrogen.

[0058] In another preferred embodiment, R3 is hydrogen or deuterium;

[0059] In another preferred embodiment, L is CH2.

[0060] In another preferred embodiment, X-A1 is -(CH2) n -O-CH2-A1.

[0061] In another preferred embodiment, n is 0.

[0062] In another preferred embodiment, A1 is halogen-substituted or unsubstituted phenyl.

[0063] In another preferred embodiment, R5 is a phenyl group, wherein the phenyl group is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, C1-C6 alkyl and halogen-substituted C1-C6 alkyl.

[0064] In another preferred embodiment, the compound has a structure shown by formula (I-12)-(I-13):

[0065] wherein R1 is hydrogen, methyl or ethyl;

[0066] R2 is hydrogen or fluorine;

[0067] R3 is hydrogen, deuterium or fluorine;

[0068] n is 0 or 1;

[0069] R6 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl;

[0070] m is 0, 1, 2, 3, 4 or 5.

[0071] In another preferred embodiment, the compound has a structure shown by formula (I-31)-(I-32):

[0072] wherein R1 is hydrogen, methyl or ethyl;

[0073] R2 is hydrogen or fluorine;

[0074] R3 is hydrogen, deuterium or fluorine;

[0075] R6 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl;

[0076] m is 0, 1, 2, 3, 4 or 5.

[0077] In another preferred embodiment, the compound has a structure shown by formula (I-20)-(I-26):

[0078] wherein R1 is hydrogen, methyl or ethyl;

[0079] R2 is hydrogen or fluorine;

[0080] R3 is hydrogen, deuterium or fluorine;

[0081] R9 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy;

[0082] p is 0, 1, 2, 3, 4, or 5;

[0083] A1 is selected from: phenyl, pyridyl, substituted or unsubstituted C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl; the aforementioned phenyl, pyridyl, substituted or unsubstituted C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl is substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy.

[0084] In another preferred embodiment, Selected from:

[0085] In another preferred embodiment, Selected from:

[0086] In another preferred embodiment, the compound represented by general formula (I) is one of the following compounds:

[0087] The compound, or a pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof.

[0088] One aspect of the present invention provides a method for preparing a compound represented by general formula (I), which is selected from one of the following methods:

[0089] The synthesis of compounds 1I and 3E is based on the following references: Organic Letters, 2020, 22(16): 6277-6282, Nature Communications, 2020, 11(1): 1-13, and Bioorg Med Chem Lett, 2013, 23(10): 2936-40.

[0090] Synthesis method 1:

[0091] Step 1-1: Compound 1A and compound 1B undergo nucleophilic substitution reaction to obtain compound 1C;

[0092] Step 1-2: Compound 1C and compound 1D undergo nucleophilic substitution reaction to obtain compound 1E;

[0093] Step 1-3: Compound 1E is hydrolyzed with base to obtain compound 1F;

[0094] Step 1-4: Compound 1F and 1G are subjected to condensation reaction to obtain compound 1H;

[0095] Step 1-5: Compound 1H reacts with tetrahydroxydiboron and sodium hydroxide to obtain compound 1I

[0096] Step 1-6: Compound 1I is reacted with p-toluenesulfonic acid under reflux to obtain compound 1J.

[0097] R5 is as defined above;

[0098] Synthesis method 2:

[0099] Step 2-1: Compound 2A reacts with compound 2B in the presence of N,N'-carbonyldiimidazole and triethylamine to obtain compound 2C;

[0100] Step 2-2: Compound 2A and compound 2D are reacted in the presence of HATU and N,N-diisopropylethylamine to obtain compound 2E.

[0101] R2, R3, R6, and m are as defined above;

[0102] Synthesis method three:

[0103] Step 3-1: Compound 3A and R8-NH2 undergo condensation reaction to obtain compound 3B;

[0104] Step 3-2: Compound 3B is hydrolyzed in the presence of lithium hydroxide to obtain compound 3C;

[0105] Step 3-3: Compound 3C and 1G are condensed to obtain compound 3D;

[0106] Step 3-4: Compound 3D reacts with tetrahydroxydiboron and sodium hydroxide to obtain compound 3E;

[0107] Step 3-5: Compound 3E is reacted with p-toluenesulfonic acid to obtain compound 3F;

[0108] R8-NH2 is selected from:

[0109] R9 is as defined above.

[0110] Another aspect of the present invention provides the use of the compounds of general formula (I) and their tautomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs, hydrates, crystalline hydrates and solvates for the preparation of drugs or diagnostic reagents for preventing or treating diseases associated with CRL4 CRBN E3 ubiquitin ligase.

[0111] Another aspect of the present invention provides the use of compounds as represented by general formula (I) and their tautomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs, hydrates thereof, crystalline hydrates, and solvates in the preparation of drugs for treating or preventing diseases, disorders, or conditions associated with abnormal production or regulation of TNF-α activity, production or regulation of IL-2 activity, or production or regulation of IFNγ activity.

[0112] Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective dose of a compound as represented by general formula (I) and its tautomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs, hydrates, crystalline hydrates, solvates and other pharmaceutically acceptable carriers.

[0113] Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective dose of a compound represented by general formula (I) and its tautomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs, hydrates, crystalline hydrates, solvates, and one or more other ingredients having pharmaceutical therapeutic activity.

[0114] The compounds of the present invention as represented by general formula (I) and their tautomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs, hydrates, crystalline hydrates, and solvates can produce a synergistic effect with one or more other ingredients having pharmaceutical therapeutic activity in the prevention or treatment of specific diseases or functional disorders.

[0115] The compounds of the present invention as represented by general formula (I) and their tautomers, enantiomers, diastereomers, racemates, metabolites, metabolic precursors, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs, hydrates, crystalline hydrates and solvates can also reduce or eliminate the toxic and side effects produced by one or more other pharmaceutically active ingredients in the prevention or treatment of specific diseases or functional disorders, and vice versa.

[0116] Another aspect of the present invention provides a pharmaceutical composition, wherein the additional one or more ingredients having pharmaceutical therapeutic activity as described above include macromolecular compounds, such as proteins, polysaccharides and nucleic acids; small molecule compounds such as inorganic compounds, organometallic compounds, organic small molecule compounds of synthetic or natural origin, etc.

[0117] Another aspect of the present invention provides a pharmaceutical composition, which is characterized in that the pharmaceutical composition further comprises other therapeutic agents, and the other therapeutic agents are one or more of dexamethasone, rituximab, trastuzumab, PD-1 inhibitors, PDL-1 inhibitors, pemetrexed, topotecan, doxorubicin, bortezomib, gemcitabine, dacarbazine, clarithromycin, vincristine, cytarabine, prednisone, docetaxel, clofarabine injection, HDAC inhibitors, androgen receptor inhibitors, androgen biosynthesis inhibitors, BTK inhibitors, erythrocyte growth hormone, minocycline, elotuzumab, palbociclib, nivolumab, pembrolizumab, panobinostat, ubliximab, romidepsin, eltrombopag, CAR-T and melphalan.

[0118] Another aspect of the present invention is to provide a compound as shown in the general formula (I) for use in the preparation of a method for treating or preventing a disease associated with CRL4 CRBN E3 ubiquitin ligase, including but not limited to cancer, pain, nervous system diseases and immune system diseases. It is characterized in that the disease, disorder or condition includes: myelodysplastic syndrome, multiple myeloma, mantle cell lymphoma, non-Hodgkin's lymphoma, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, myelofibrosis, Burkitt's lymphoma, Hodgkin's lymphoma, large cell lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, ciliary body and chronic melanoma, iris melanoma, recurrent interocular melanoma, T cell lymphoma, erythroid lymphoma, monoblastic and monocytic leukemia. , myeloid leukemia, central nervous system lymphoma, brain tumor, meningioma, spinal cord tumor, thyroid cancer, non-small cell lung cancer, ovarian cancer, skin cancer, renal cell carcinoma, astrocytoma, amyloidosis, complex regional pain syndrome type I, malignant melanoma, radiculopathy, myelofibrosis, glioblastoma, gliosarcoma, malignant glioma, refractory plasmacytoma, melanoma with extraocular extension, solid tumors, papillary and follicular thyroid cancer, breast cancer, prostate cancer, hepatocellular carcinoma, or essential macroglobulinemia.

[0119] Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by formula (I), and one or more of its stereoisomers, pharmaceutically acceptable salts, prodrugs, solvates, hydrates, and crystalline forms, and at least one excipient, diluent, or carrier. A typical formulation is prepared by mixing the compound represented by formula (I) of the present invention with a carrier, diluent, or excipient. Suitable carriers, diluents, or excipients are well known to those skilled in the art and include substances such as carbohydrates, waxes, water-soluble and / or swellable polymers, hydrophilic or hydrophobic substances, gelatin, oils, solvents, and water. The specific carrier, diluent, or excipient used will depend on the intended use and purpose of the compound of the present invention. Solvents are generally selected based on solvents deemed safe and effective for administration to mammals by those skilled in the art. Generally, safe solvents are non-toxic aqueous solvents such as water, as well as other non-toxic solvents that are soluble or miscible in water. Suitable aqueous solvents include one or more of water, ethanol, propylene glycol, polyethylene glycol (such as PEG400, PEG300), etc. The formulation may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavorings, flavorings or other known additives to enable the drug to be manufactured or used in an acceptable form.

[0120] When the compound of formula (I) of the present invention is used in combination with at least one other drug, the two or more drugs can be used separately or in combination, preferably administered in the form of a pharmaceutical composition. The compound of formula (I) or pharmaceutical composition of the present invention can be administered to a subject separately or together in any known oral, intravenous, rectal, vaginal, transdermal, or other topical or systemic administration form.

[0121] These pharmaceutical compositions may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavorings, flavorings or other known additives so that the pharmaceutical compositions can be manufactured or used in an acceptable form.

[0122] The medicament of the present invention is preferably administered orally. Solid dosage forms for oral administration may include capsules, tablets, powders, or granular formulations. In a solid dosage form, the compound or pharmaceutical composition of the present invention is mixed with at least one inert excipient, diluent, or carrier. Suitable excipients, diluents, or carriers include substances such as sodium citrate or dicalcium phosphate, or starch, lactose, sucrose, mannitol, silicic acid, etc.; binders such as carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.; humectants such as glycerol, etc.; disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, specific complex silicates, sodium carbonate, etc.; solution retardants such as paraffin, etc.; absorption enhancers such as quaternary ammonium compounds, etc.; adsorbents such as kaolin, bentonite, etc.; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc. In the case of capsules and tablets, the dosage form may also include a buffer. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using, for example, lactose and high molecular weight polyethylene glycols as excipients.

[0123] The liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the compound of the present invention or its composition, the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvents; solubilizers and emulsifiers such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide; oils (such as cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, etc.); glycerol; tetrahydrofurfuryl alcohol; fatty acid esters of polyethylene glycol and sorbitan; or mixtures of several of these substances.

[0124] In addition to these inert diluents, the composition may also include excipients such as one or more of a wetting agent, an emulsifier, a suspending agent, a sweetener, a flavoring agent, and a fragrance. In the case of a suspension, in addition to the compound or combination of the present invention, it may further contain a carrier such as a suspending agent, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and tragacanth, or a mixture of several of these substances.

[0125] Compositions for rectal or vaginal administration are preferably suppositories and can be prepared by mixing the compound or combination of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at room temperature but liquid at body temperature and melt in the rectum or vagina to release the active compound.

[0126] The compounds or pharmaceutical compositions of the present invention may be administered in other topical dosage forms, including creams, powders, sprays, and inhalants. The drug may be mixed under sterile conditions with a pharmaceutically acceptable excipient, diluent, or carrier, and any desired preservatives, buffers, or propellants. Ophthalmic formulations, ophthalmic ointments, powders, and solutions are also intended to be encompassed within the scope of this invention.

[0127] The present invention also provides compounds represented by the general formula (I) and their tautomers, enantiomers, diastereomers, racemates, metabolic precursors, metabolites, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs or hydrates, solvates or crystal forms thereof as CRL4 CRBN Use of selective modulators of E3 ubiquitin ligases to regulate CUL4 CRBN E3 ubiquitin ligase activity.

[0128] The present invention also provides a compound as shown in the general formula (I) and its tautomers, enantiomers, diastereomers, racemates, metabolic precursors, metabolites, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs or hydrates, solvates or crystal forms thereof, which can be used in the preparation of a compound for treating or preventing CRL4. CRBN Use of drugs for diseases related to E3 ubiquitin ligase involvement. CRBN Diseases related to E3 ubiquitin ligase involvement include (but are not limited to) tumors, central nervous system diseases, and immune diseases.

[0129] In a preferred embodiment, the present invention relates to a method for treating or preventing a disease, disorder or condition associated with TNF-α production or regulation of TNF-α activity, IL-2 production or dysregulation of IL-2 activity, wherein the method comprises administering to a subject an effective amount of an isoindoline derivative represented by general formula (I), a pharmaceutically acceptable salt, solvate, stereoisomer, isotope compound, metabolite and prodrug thereof. According to the method of the present invention, examples of such diseases, disorders and conditions to be treated or prevented include, but are not limited to, cancer, including solid tumors, TNF-α-related disorders, undesirable angiogenesis-related diseases and disorders, pain, macular degeneration (MD)-related syndromes, skin diseases, keratosis, respiratory diseases (e.g., lung diseases), immunodeficiency diseases, central nervous system (CNS) diseases, autoimmune diseases, atherosclerosis, genetic diseases, allergies, viruses, sleep disorders and related syndromes, inflammatory diseases, PDE-4-related diseases or IL-2-related diseases. Examples of such diseases, disorders or conditions that are well known in the art include, but are not limited to, those described in PCT Patent Publications WO2012015986 and WO2006018182, and US Patent Publication US20100204227, some of which are hereby incorporated by reference in their entirety.

[0130] The compounds represented by the general formula (I) of the present invention, and their stereoisomers, pharmaceutically acceptable salts, prodrugs, solvates, hydrates or crystalline forms can be used in monotherapy or combination therapy. When used in combination therapy, the compounds of formula (I), their enantiomers, diastereomers, racemates and mixtures thereof, as well as their pharmaceutically acceptable salts, crystalline hydrates and solvates, are included in a therapeutically effective dose, and one or more other ingredients with pharmaceutical therapeutic activity. The one or more other ingredients with pharmaceutical therapeutic activity include macromolecular compounds, such as proteins (antibodies or polypeptides), polysaccharides and nucleic acids (DNA or RNA), etc.; and small molecule compounds, such as inorganic compounds, organometallic compounds, synthetic or naturally derived organic small molecule compounds, etc.; in addition, radiation, surgery, cell therapy, hormone therapy or cytokine therapy, etc. are also included. The compounds of formula (I) of the present invention, their prodrugs, enantiomers, diastereomers, racemates, and mixtures thereof, as well as their pharmaceutically acceptable salts, crystalline hydrates, and solvates, can produce a synergistic effect with one or more other ingredients having pharmaceutical therapeutic activity in the prevention or treatment of specific diseases or functional disorders. The compounds of formula (I) of the present invention, their prodrugs, enantiomers, diastereomers, racemates, and mixtures thereof, as well as their pharmaceutically acceptable salts, crystalline hydrates, and solvates, can also reduce or eliminate the toxic side effects produced by one or more other ingredients having pharmaceutical therapeutic activity in the prevention or treatment of specific diseases or functional disorders, and vice versa.

[0131] In another preferred embodiment, the disease or functional disorder includes, but is not limited to: cancer, diseases or functional disorders related to angiogenesis, pain (including but not limited to complex regional pain syndrome), macular degeneration and related functional disorders, skin diseases, lung functional disorders, immunodeficiency diseases, damage and functional disorders of the central nervous system, and TNFα-related diseases or functional disorders.

[0132] In another preferred embodiment, the cancer includes, but is not limited to, skin cancer (such as melanoma), lymphatic system cancer, breast cancer, cervical cancer, uterine cancer, digestive tract cancer, lung cancer, ovarian cancer, prostate cancer, colon cancer, rectal cancer, oral cancer, brain tumor, head and neck cancer, pharyngeal cancer, testicular cancer, kidney cancer, pancreatic cancer, spleen cancer, liver cancer, bladder cancer, laryngeal cancer, and cancers associated with AIDS. The compounds provided by the present invention are also effective for hematologic malignancies and myelomas, such as multiple myeloma and acute and chronic leukemia. The compounds provided by the present invention can be used to prevent or treat primary tumors and metastatic tumors.

[0133] The compounds represented by the general formula (I) may contain one or more asymmetric or chiral centers and may therefore exist in different stereoisomeric forms. The compounds of the present invention include all stereoisomeric forms, including but not limited to diastereomers, enantiomers and atropisomers and mixtures thereof (such as racemic mixtures), metabolic precursors, metabolites, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof. The compounds represented by the general formula (I) may also exist in different tautomeric forms, all of which are included within the scope of the present invention.

[0134] 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 in the embodiments) can be combined with each other to form new or preferred technical solutions. The above description is not intended to limit the aspects of the present invention in any form. DETAILED DESCRIPTION

[0135] After extensive and in-depth research, the present inventors have provided a class of CRL4 with novel structure and excellent anti-tumor activity through a large number of screening and testing. CRBN E3 ubiquitin ligase modulator compounds. The inventors of the present invention have studied CRL4 CRBN The following important information was obtained by analyzing the crystal structure of the complex between E3 ubiquitin ligase and small molecules (PDB ID: 4CI2, 5HXB): CRL4 CRBN E3 ubiquitin ligases have multiple binding pockets for small molecules, so it is possible to develop small molecules with complex structures and multiple binding sites to achieve CRL4 CRBN E3 ubiquitin ligase effectively binds to small molecules. Molecular dynamics simulation methods are used to analyze the structural dynamics and binding site of the model molecule and E3 ubiquitin ligase interface. Combined with molecular docking and complex-based pharmacophore matching strategies, the binding mode and interaction of the compound at the E3 ubiquitin ligase active site are evaluated through scoring functions, and computational simulation and optimization of structural design are performed to obtain a new specific CRL4 CRBN Small molecule modulators of E3 ubiquitin ligases.

[0136] Based on this information, the inventors designed and synthesized a series of CRL4 with novel indazolone skeleton structure described in this application. CRBNSmall molecule modulators of E3 ubiquitin ligases were developed, and the activity of the compounds was tested. The test results of representative compounds in multiple myeloma cell lines (MM.1S), mantle cell lymphoma cell lines (Mino), and acute myeloid leukemia cell lines (MV-4-11) showed that the new small molecule modulators have very high cell growth inhibitory activity. However, the original CRBN modulators with isoindoline structures, that is, IMiDs that are already on the market or under clinical development, do not have good cell growth inhibitory activity in mantle cell lymphoma cell lines (Mino) or acute myeloid leukemia cell lines (MV-4-11). Therefore, after the indazolone compounds developed in this application act on the organism, they can regulate the degradation of substrate proteins or regulate the degradation of new pathogenic proteins by regulating the ubiquitin-proteasome-mediated protein degradation pathway in the organism, thereby achieving effective disease treatment based on the CRBN target. The present invention was completed on this basis.

[0137] the term

[0138] When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents that would result if the formula were written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0139] Those skilled in the art will understand that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible. References to substitutions in the relevant structures herein include both substitution and non-substitution, such as "optionally" or "optionally" substituted with a substituent, to including both substitution and non-substitution.

[0140] The term "when the number of substituents is greater than 1, the R substituents may be the same or different substituents" mentioned in the present invention means that when there are multiple substituents in a certain structure, the substituent combination of R may be selected from a variety of different types of substituents.

[0141] The term "substituted" only applies to sites that can be substituted by substituents, and does not include substitutions that cannot be achieved based on existing chemical knowledge. The term "substituted" refers to the replacement of one or more hydrogen atoms on a specific group by a specific substituent. The specific substituents are the substituents described above, or the substituents that appear in the examples. Unless otherwise specified, an arbitrarily substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituents may be the same or different at each position. A cyclic substituent, such as a heterocycloalkyl, may be connected to another ring, such as a cycloalkyl, to form a spirobicyclic system, for example, where the two rings share one carbon atom. Unless otherwise specified, substitution may refer to a group being substituted by one or more groups selected from the group consisting of hydrogen, halogen, cyano, nitro, hydroxy, carboxyl, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, and C1-C6 alkoxy-substituted C1-C6 alkoxy.

[0142] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0143] The term "plurality" refers to two or more, such as 2, 3, 4, 5 or 6.

[0144] The term "C1-C6 alkyl" refers to a straight or branched saturated hydrocarbon group having 1 to 6 carbon atoms in the chain, such as an alkyl group containing 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group used in this article or as part of another group has 1 to 6 carbon atoms (i.e., C1-C6 alkyl), or 1 to 3 carbon atoms (i.e., C1-3 alkyl), including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl and the like.

[0145] The term "C1-C6 alkoxy" refers to a straight or branched chain alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy, butoxy and the like.

[0146] The term "aryl" refers to a 6-14 membered all-carbon monocyclic or fused polycyclic group having a conjugated p-electron system, preferably a 6- to 10-membered ring (i.e., C6-C10 aryl), more preferably phenyl and naphthyl, and most preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring.

[0147] The term "halogen-substituted C1-C6 alkoxy" refers to a linear or branched C1-C6 alkoxy group substituted by a single or multiple halogens, including but not limited to -OCH2F, -OCHF2, and -OCF3.

[0148] The term "halogen-substituted C1-C6 alkyl" refers to a straight-chain or branched C1-C6 alkyl group substituted by a single or multiple halogens, including but not limited to 2-bromoethyl, 2-bromopropyl, and the like.

[0149] The term "C3-C10 cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent. The C3-C10 cycloalkyl is preferably a C4-C7 cycloalkyl or a C5-C6 cycloalkyl. Monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentenyl, and cyclohexyl. Polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls.

[0150] The term "4-10 membered heterocyclyl" refers to a group containing one or more saturated and / or partially saturated rings, which includes 4 to 10 ring atoms, wherein one or more (e.g., 2, 3) ring atoms are selected from nitrogen, oxygen, sulfur or S(O)m (wherein m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. The 4-10 membered heterocyclyl is preferably a 4-6 membered heterocyclyl, for example, propylene oxide, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl.

[0151] The term "C2-C6 alkenyl" means a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having 2 to 6 carbon atoms and connected to the rest of the molecule by one or more single bonds, such as, but not limited to, ethenyl, propenyl, butenyl, styryl, and phenylpropenyl.

[0152] The term "heteroaryl" refers to a 5-14 membered aromatic group having 1 to 4 (e.g., 2 or 3) heteroatoms as ring atoms, with the remaining ring atoms being carbon, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen, preferably 5-10 members, more preferably 5 or 6 members, such as thienyl, furyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like.

[0153] "Hydrate" refers to a compound containing water.

[0154] "Pharmaceutically acceptable salts" refer to salts formed between the drug molecule and the corresponding organic or inorganic acid or organic or inorganic base. Inorganic acids from which salts can be derived include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, trifluoroacetic acid, succinic acid, mesylate, and the like. By way of example only, salts derived from inorganic bases include sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, such as alkylamines (i.e., NH2(alkyl)), dialkylamines (i.e., HN(alkyl)2), trialkylamines (i.e., N(alkyl)3), substituted alkylamines (i.e., NH2(substituted alkyl)), di(substituted alkyl)amines (i.e., HN(substituted alkyl)2), tri(substituted alkyl)amines (i.e., N(substituted alkyl)3), etc. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(isopropyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.

[0155] In the present invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art. In the present invention, the definitions of different general formulas or multiple identical groups in the same general formula are independent of each other.

[0156] Active ingredient

[0157] As used herein, "compound of the present invention" or "active ingredient" refers to the compound represented by Formula I, and also includes pharmaceutically acceptable salts, tautomers, stereoisomers, and prodrugs thereof.

[0158] The compound represented by formula (I) may also exist in different tautomeric forms, all of which are included within the scope of the present invention.

[0159] The term "tautomer" refers to constitutional isomers of different energies that are interconvertible via a low energy barrier, the reaction typically resulting in the formal migration of a hydrogen atom or proton accompanied by conversion of a single bond and an adjacent double bond.

[0160] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. All such compounds contemplated by the present invention, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, racemates, and other mixtures, are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and their mixtures are encompassed by the present invention.

[0161] The compounds of formula (I) may contain one or more asymmetric or chiral centers and may therefore exist in different stereoisomeric forms. The compounds of the present invention include all stereoisomeric forms, including but not limited to diastereomers, enantiomers and atropisomers and mixtures thereof (e.g., racemates), which are all included within the scope of the present invention.

[0162] Unless otherwise indicated, the term "enantiomer" refers to stereoisomers that are mirror images of one another.

[0163] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0164] "Racemate" refers to two stereoisomers that are mirror images of each other, with opposite optical rotations that cancel each other out.

[0165] There are chiral centers in the general formula and example compounds of the present invention. Therefore, the single configuration or racemate of the compound of the present invention is also within the scope of this application. In addition, there is corresponding evidence in the prior art that compounds with an amine structure, their R configuration, S configuration, and racemate will all be converted into a 1:1 racemate on cells in in vitro cell experiments and enter the body. Therefore, those skilled in the art can expect that the technical solutions corresponding to the enantiomers or racemates of the compound of the present invention can achieve comparable technical effects. If it is necessary to obtain an example compound of a single configuration in the present invention, the example compound of a single configuration can be obtained through the intermediate 1B of a single configuration and the synthetic method 1 in the compound preparation method, thereby obtaining an enantiomer. In addition, the example compound of a diastereoisomer can be obtained by preparing the intermediate 1B of a single configuration, so diastereoisomers can also be obtained in the present invention. These different isomeric forms are all stereoisomers.

[0166] The compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more atoms comprising the compound. For example, the compound may be labeled with radioactive isotopes, such as deuterium (D), tritium (T), 12C, 13C, and 14C. For another example, deuterated drugs may be formed by replacing hydrogen with heavy hydrogen. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared to non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced therapeutic efficacy, and extended drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

[0167] As used herein and unless otherwise specified, the term "prodrug" refers to a derivative of a compound that contains a bioreactive functional group such that the bioreactive functional group can be cleaved from the compound or otherwise reacted under biological conditions (in vitro or in vivo) to provide the compound. Typically, the prodrug is inactive, or at least less active than the compound itself, such that its activity is not exerted until the compound is cleaved from the bioreactive functional group. The bioreactive functional group can be hydrolyzed or oxidized under biological conditions to provide the compound. For example, a prodrug may contain a biohydrolyzable group. Examples of biohydrolyzable groups include, but are not limited to, biohydrolyzable phosphates, biohydrolyzable esters, biohydrolyzable amides, biohydrolyzable carbonates, biohydrolyzable carbamates, and biohydrolyzable ureides. When the compounds of the present invention contain basic groups, they can be prepared as pharmaceutically acceptable salts, including inorganic and organic acid salts. 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, and phosphoric acid; organic acids such as 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, toluenesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

[0168] The present invention also encompasses any novel intermediates disclosed herein.

[0169] Pharmaceutical compositions and methods of administration

[0170] As used herein, a "pharmaceutical composition" refers to a formulation of a compound of the present invention and a medium generally accepted in the art for delivering a biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration to an organism, thereby facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0171] The compounds of formula (I) can be used in combination with other drugs known to treat or improve similar conditions. When administered in combination, the original drug's route of administration and dosage can remain unchanged, while the compound of formula I is taken simultaneously or subsequently. When the compound of formula I is taken simultaneously with one or more other drugs, a pharmaceutical composition containing one or more known drugs and the compound of formula I can be preferably used. Drug combinations also include taking the compound of formula I and one or more other known drugs during overlapping time periods. When the compound of formula I is used in combination with one or more other drugs, the dosage of the compound of formula I or the known drug may be lower than the dosage of each drug taken alone.

[0172] As used herein, the term "pharmaceutically acceptable" refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, that is, the substance can be administered to a subject without causing an adverse biological response or interacting in an adverse manner with any components contained in the composition.

[0173] In this application, "pharmaceutically acceptable carrier" includes but is not limited to any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or livestock use.

[0174] Drugs or active ingredients that can be used in combination with the compound of general formula (I) include but are not limited to one or more of dexamethasone, rituximab, trastuzumab, PD-1 inhibitors, PDL-1 inhibitors, pemetrexed, topotecan, doxorubicin, bortezomib, gemcitabine, dacarbazine, clarithromycin, vincristine, cytarabine, prednisone, docetaxel, clofarabine injection, HDAC inhibitors, kinase targeted inhibitors, androgen receptor inhibitors, androgen biosynthesis inhibitors, erythrocyte growth hormone, minocycline, elotuzumab, palbociclib, nivolumab, pembrolizumab, panobinostat, ublizumab, romidepsin, eltrombopag, CAR-T and melphalan.

[0175] A typical formulation is prepared by mixing a compound represented by the general formula (I) of the present invention and a carrier, diluent or excipient. Suitable carriers, diluents or excipients are well known to those skilled in the art and include substances such as carbohydrates, waxes, water-soluble and / or swellable polymers, hydrophilic or hydrophobic substances, gelatin, oils, solvents, water, etc. The specific carrier, diluent or excipient used will depend on the mode of use and purpose of the compound of the present invention. Solvents are generally selected based on solvents that are considered safe and effective for administration to mammals by those skilled in the art. In general, safe solvents are non-toxic aqueous solvents such as pharmaceutical water, and other non-toxic solvents that are soluble in water or miscible with water. Suitable aqueous solvents include one or more of water, ethanol, propylene glycol, polyethylene glycol (such as PEG400, PEG300), etc. The formulation may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavorings, flavorings or other known additives to enable the compound represented by formula (I) to be manufactured or used in an acceptable form.

[0176] When the compound of formula (I) of the present invention is used in combination with at least one other drug, the two or more drugs can be used separately or in combination, preferably administered in the form of a pharmaceutical composition. The compound of formula (I) or pharmaceutical composition of the present invention can be administered to a subject separately or together in any known oral, intravenous, rectal, vaginal, transdermal, or other topical or systemic administration form.

[0177] These pharmaceutical compositions may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavorings, flavorings or other known additives so that the pharmaceutical compositions can be manufactured or used in an acceptable form.

[0178] The medicament of the present invention is preferably administered orally. Solid dosage forms for oral administration may include capsules, tablets, powders, or granular formulations. In a solid dosage form, the compound or pharmaceutical composition of the present invention is mixed with at least one inert excipient, diluent, or carrier. Suitable excipients, diluents, or carriers include substances such as sodium citrate or dicalcium phosphate, or starch, lactose, sucrose, mannitol, silicic acid, etc.; binders such as carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.; humectants such as glycerol, etc.; disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, specific complex silicates, sodium carbonate, etc.; solution retardants such as paraffin, etc.; absorption enhancers such as quaternary ammonium compounds, etc.; adsorbents such as kaolin, bentonite, etc.; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc. In the case of capsules and tablets, the dosage form may also include a buffer. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using, for example, lactose and high molecular weight polyethylene glycols as excipients.

[0179] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the compound of the present invention or its composition, the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvents; solubilizers and emulsifiers such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide; oils (such as cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, etc.); glycerol; tetrahydrofurfuryl alcohol; fatty acid esters of polyethylene glycol and sorbitan; or mixtures of several of these substances.

[0180] In addition to these inert diluents, the composition may also include excipients such as one or more of wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, and perfuming agents.

[0181] In the case of suspensions, in addition to the compound or combination of the present invention, it may further contain a carrier such as a suspending agent, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, or a mixture of several of these substances.

[0182] Compositions for rectal or vaginal administration are preferably suppositories and can be prepared by mixing the compound or combination of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at room temperature but liquid at body temperature and melt in the rectum or vagina to release the active compound.

[0183] The compounds or pharmaceutical compositions of the present invention may be administered in other topical dosage forms, including creams, powders, sprays, and inhalants. The drug may be mixed under sterile conditions with a pharmaceutically acceptable excipient, diluent, or carrier, and any desired preservatives, buffers, or propellants. Ophthalmic formulations, ophthalmic ointments, powders, and solutions are also intended to be encompassed within the scope of this invention.

[0184] As used herein, the terms "administer," "administer," "administer," and the like refer to methods by which a compound or composition can be delivered to the desired site for biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injections or infusions), topical administration, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used for the compounds and methods described herein, such as those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

[0185] As used herein, the terms "pharmaceutical combination," "drug combination," "combination therapy," "administration of another therapy," "administration of another therapeutic agent," and the like refer to a pharmaceutical therapy obtained by mixing or combining more than one active ingredient, and include both fixed and non-fixed combinations of the active ingredients. The term "fixed combination" means that at least one compound described herein and at least one synergistic agent are administered to a patient simultaneously as a single entity or single dosage form. The term "non-fixed combination" means that at least one compound described herein and at least one synergistic agent are administered to a patient simultaneously, in combination, or sequentially at variable intervals as separate entities. This also applies to cocktail therapies, e.g., administration of three or more active ingredients.

[0186] The pharmaceutical composition of the present invention comprises a safe and effective amount of a compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0187] The compound or pharmaceutical composition of the present invention can be used to prepare a drug for treating or preventing diseases in which CRL4CRBNE3 ubiquitin ligase participates. The diseases related to CRL4CRBNE3 ubiquitin ligase participates described in the present invention include, but are not limited to, tumors, central nervous system diseases, and immune diseases.

[0188] In a preferred embodiment, the disease includes but is not limited to: cancer, pain (including but not limited to complex regional pain syndrome), skin diseases, immunodeficiency diseases, central nervous system damage and dysfunction.

[0189] In another preferred embodiment, the cancer includes (but is not limited to): skin cancer (such as melanoma), lymphatic system cancer, breast cancer, cervical cancer, uterine cancer, digestive tract cancer, lung cancer, ovarian cancer, prostate cancer, colon cancer, rectal cancer, oral cancer, brain tumor, head and neck cancer, pharyngeal cancer, testicular cancer, kidney cancer, pancreatic cancer, spleen cancer, liver cancer, bladder cancer, laryngeal cancer and cancer related to AIDS. The compounds provided by the present invention are also effective for hematological tumors and myeloma, such as multiple myeloma, lymphoma and acute and chronic leukemia. The compounds provided by the present invention can also be used to prevent or treat primary tumors and metastatic tumors.

[0190] The use of the drug molecules of the present invention includes but is not limited to the diseases selected from the group consisting of myelodysplastic syndrome, multiple myeloma, mantle cell lymphoma, non-Hodgkin's lymphoma, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, myelofibrosis, Burkitt's lymphoma, Hodgkin's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, ciliary body and chronic melanoma, iris melanoma, recurrent interocular melanoma, T cell lymphoma, erythroid lymphoma, monoblastic leukemia, Myeloid and monocytic leukemia, myeloid leukemia, central nervous system lymphoma, meningioma, spinal cord tumor, non-small cell lung cancer, ovarian cancer, skin cancer, renal cell carcinoma, astrocytoma, amyloidosis, complex regional pain syndrome type I, malignant melanoma, radiculopathy, glioblastoma, gliosarcoma, malignant glioma, refractory plasmacytoma, melanoma with extraocular extension, papillary and follicular thyroid cancer, breast cancer, prostate cancer, hepatocellular carcinoma, or essential macroglobulinemia.

[0191] The main advantages of the present invention include:

[0192] 1. The present invention provides a novel indazolone skeleton structure CRL4 CRBN E3 ubiquitin ligase molecular glue degraders have improved tumor therapeutic effects and can broaden the indications of molecular glue degraders, and provide CRL4 with better physicochemical properties for the design and development of degraders of related pathogenic proteins. CRBN Ligand for E3 ubiquitin ligase.

[0193] 2. Based on the structural instability of existing CRBN ligands and immunomodulatory imide drugs (IMiDs), which are easily hydrolyzed in body fluids, this invention optimizes their structure and develops a molecular glue-based degradation agent with an indazolone skeleton (Formula I) through skeleton transition. This skeleton has good chemical stability.

[0194] The present invention can quickly and efficiently construct the indazolone molecular glue skeleton shown in formula I through light-induced cyclization of primary amine and o-nitrobenzyl alcohol (PANAC reaction).

[0195] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0196] In all embodiments, 1H NMR spectra were measured using 400 MHz Bruker and 500 MHz Bruker nuclear magnetic resonance instruments, with chemical shifts expressed in δ (ppm). Mass spectra were determined using MS UPLC-MS (ESI). The UPLC model was Waters HPLC H-CLASS, and the MS (ESI) model was Waters SQ Detector 2. Anhydrous tetrahydrofuran was prepared by reflux drying and deoxygenation of benzophenone / sodium metal, and anhydrous toluene and anhydrous dichloromethane were prepared by reflux drying of calcium chloride. Solvents used for column chromatography mobile phases, such as petroleum ether, ethyl acetate, and dichloromethane, were purchased from Sinopharm Chemical Reagent Co., Ltd. The thin layer chromatography silica gel plates (HSGF254) used in reaction detection were from Sinopharm Chemical Reagent Co., Ltd. Compound separation used 200-300 mesh silica gel from Sinopharm Chemical Reagent Co., Ltd. The raw materials in the present invention can be obtained from commercial sources, such as the main reagents purchased from Sinopharm Chemical Reagent Co., Ltd., or prepared by methods known in the art, or prepared according to the methods described in the present invention.

[0197] Example 1: (S)-3-(7-((4-(morpholinomethyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D1)

[0198] Synthesis route:

[0199] Step 1: A suspension of compound 1.1 (500 mg, 2.54 mmol), compound 1.2 (2.0 g, 7.60 mmol) and potassium carbonate (351 mg, 2.54 mmol) in acetonitrile was heated to 50°C for overnight reaction. After completion of the reaction as monitored by TLC, the suspension was cooled to room temperature, filtered, and concentrated under reduced pressure. Column chromatography afforded 910 mg of a white solid (yield 94%). 1 H NMR (500MHz, CDCl3) δ7.61 (dd, J=7.9, 1.1Hz, 1H), 7.47–7.39 (m, 3H), 7.34 (d, J= 8.2Hz,2H),7.24(dd,J=8.4,1.0Hz,1H),5.20(s,2H),4.48(s,2H),3.90(s,3H).

[0200] Step 2: A suspension of compound 1.3 (350 mg, 0.9 mmol), morpholine (86 μL, 1.01 mmol) and potassium carbonate (191 mg, 1.38 mmol) in acetonitrile was stirred at room temperature overnight. After completion of the reaction as monitored by TLC, the mixture was filtered, concentrated under reduced pressure, and purified by column chromatography to give 351 mg of a white solid (yield 99%).

[0201] Step 3: Dissolve compound 1.4 (393 mg, 1.0 mmol) in 10 mL of anhydrous tetrahydrofuran, add 5 mL of lithium hydroxide (63 mg, 1.5 mmol) aqueous solution, stir at room temperature for 2 hours, and after completion of the reaction as monitored by TLC, concentrate under reduced pressure and dry in vacuo for direct use in the next step.

[0202] Step 4: Compound 1.5 (201 mg, 0.52 mmol), (S)-tert-butyl 4,5-diamino-5-oxopentanoate (124 mg, 0.52 mmol), and HATU (198 mg, 0.52 mmol) were dissolved in 10 mL of anhydrous DMF. DIPEA (258 μL, 1.56 mmol) was added at room temperature. The reaction system was allowed to react overnight at room temperature. After TLC showed that the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic layer was separated and washed several times with saturated sodium chloride solution, dried, filtered, concentrated, and subjected to column chromatography to obtain 210 mg of a gray solid with a yield of 73%. The theoretical value of UPLC-MS (ESI) was C 28 H 36 N4O8[M+H] + :557.62, measured value: 557.62.

[0203] Step 5: Compound 1.6 (202 mg, 0.36 mmol) was dissolved in methanol (5 mL). Tetrahydroxydiboron (161 mg, 1.8 mmol) was added at 0°C, followed by a methanol solution (5 mL) of sodium hydroxide (144 mg, 3.6 mmol). The reaction was continued at the same temperature for 10 minutes, and then the temperature was raised to 40°C for overnight reaction. After TLC monitoring, the reaction was completed, and saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate was added for extraction, and the layers were separated. The organic phase was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography to obtain 160 mg of the product with a yield of 85%. The theoretical value of UPLC-MS (ESI) was C 28 H 36 N4O6[M+H] + :525.62, measured value: 525.53. 1H NMR (500MHz, DMSO) δ9.75 (s, 1H), 7.48 (t, J = 9.4Hz, 3H), 7.34 (d, J = 7.9Hz, 2H), 7. 30(s,1H),7.23(d,J=7.8Hz,1H),7.18(d,J=7.9Hz,1H),7.05(t,J=7.8Hz,1H),5.2 2(s,2H),4.87(dd,J=9.7,6.0Hz,1H),3.63–3.51(m,4H),3.47(s,2H),2.94(d,J= 29.3Hz,2H),2.34(s,4H),2.20(d,J=14.5Hz,2H),2.12–1.97(m,2H),1.35(s,9H).

[0204] Step 6: Compound 1.7 (75 mg, 0.14 mmol) and p-toluenesulfonic acid monohydrate (160 mg, 0.14 mmol) were dissolved in acetonitrile (6 mL) and reacted at 80°C overnight. The reaction solution was concentrated and purified by HPLC to obtain 60 mg of the product with a yield of 95%. The theoretical value of UPLC-MS (ESI) was C 24 H 26 N4O5[M+H] + :451.50, measured value: 451.53. 1 H NMR (500MHz, DMSO) δ10.98(s,1H),10.09(s,1H),7.49(d,J=8.0Hz,2H),7.34(d,J=8.0 Hz,2H),7.25(d,J=7.7Hz,1H),7.20(d,J=7.8Hz,1H),7.05(t,J=7.8Hz,1H),5.30(dd, J=13.1,5.2Hz,1H),5.22(s,2H),3.60–3.53(m,4H),3.47(s,2H),2.92(ddd,J=17.2,1 3.8,5.4Hz,1H),2.65–2.58(m,1H),2.57–2.51(m,1H),2.34(s,4H),2.10–2.03(m,1H). 13 C NMR (126MHz, DMSO) δ172.78,169.63,165.05,145.19,137.75,135.21,132.16,129.00,127.91,1 21.77,118.55,116.66,114.81,112.95,69.48,66.20,62.14,53.87,53.16,48.61,30.94,22.36.

[0205] Example 2: (S)-4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazol-7-yl)oxy)methyl))benzyl)piperazin-1-yl)-3-fluorobenzonitrile (D2)

[0206] Synthesis route:

[0207] Step 1: Dissolve methyl 3-((4-(bromomethyl)benzyl)oxy)-2-nitrobenzoate (321 mg, 0.844 mmol), 3-fluoro-4-piperazinylbenzonitrile (191 mg, 0.93 mmol) and potassium carbonate (175 mg, 1.27 mmol) in acetonitrile (15 mL) and react at room temperature overnight. After TLC monitoring, the reaction was completed and filtered. The filtrate was concentrated and column chromatography was performed to obtain 308 mg of the product as a white solid in a yield of 72%. UPLC-MS (ESI) theoretical value was C 27 H 25 FN4O5[M+H] + :505.52, measured value: 505.57.

[0208] Step 2: Dissolve the compound 3-((4-((4-(4-cyano-2-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-2-nitrobenzoic acid methyl ester (345 mg, 0.68 mmol) in tetrahydrofuran (8 mL), add lithium hydroxide monohydrate (43 mg, 1.03 mmol) in water (8 mL), stir at room temperature and react overnight. The reaction solution was concentrated under reduced pressure and dried to obtain 315 mg of the product, which was used directly in the next reaction without purification.

[0209] Step 3: Compound 3-((4-((4-(4-cyano-2-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-2-nitrobenzoic acid lithium salt (315 mg, 0.68 mmol), (S)-tert-butyl 4,5-diamino-5-oxopentanoate monohydrochloride (162 mg, 0.68 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (259 mg, 0.68 mmol) were added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.332 mL, 2.04 mmol) was added dropwise. The reaction mixture was reacted at room temperature for 12 hours. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain 423 mg of the product as a yellow oil with a yield of 98%. The theoretical value of UPLC-MS (ESI) is C 35 H 39 FN6O7[M+H] +:675.73, measured value: 675.61.

[0210] Step 4: Compound (S)-tert-butyl 5-amino-4-(3-((4-((4-(4-cyano-2-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-2-nitrobenzamido)-5-oxopentanoate (421 mg, 0.62 mmol) was added to methanol (10 mL), tetrahydroxydiboron (278 mg, 3.1 mmol) was added at 0°C, and then a methanol solution of sodium hydroxide (248 mg, 6.2 mmol) (10 mL) was added. The mixture was reacted at 0°C for 10 minutes, and then the temperature was raised to 40°C and reacted overnight. After the reaction was complete as monitored by TLC, a saturated ammonium chloride solution was added to quench the reaction, and ethyl acetate was added for extraction. The organic phase was separated, washed with water and a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography to obtain 265 mg of the product as a colorless oil with a yield of 66%.

[0211] Step 5: Compound (S)-5-amino-4-(7-((4-((4-(4-cyano-2-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)-5-oxopentanoic acid tert-butyl ester (265 mg, 0.41 mmol) and p-toluenesulfonic acid monohydrate (471 mg, 2.47 mmol) were added to acetonitrile (8 mL) and reacted at 80°C overnight. The reaction solution was concentrated and purified by HPLC to obtain 47 mg of the product with a yield of 20%. The theoretical value of UPLC-MS (ESI) was C 31 H 29 FN6O4[M+H] + :569.61, measured value: 569.55. 1 H NMR (600MHz, DMSO-d6) δ10.97(s,1H),10.10(s,1H),7.68(dd,J=13.3,1.5Hz,1H),7.56(dd,J=8.5,1 .6Hz,1H),7.50(d,J=7.7Hz,2H),7.37(d,J=7.6Hz,2H),7.25(d,J=7.8Hz,1H),7.20(d,J=7.8Hz,1H) ,7.11(s,1H),7.06(t,J=7.8Hz,1H),5.30(dd,J=13.1,5.2Hz,1H),5.23(s,2H),3.55(s,2H),3.18(s ,4H),2.92(ddd,J=17.3,13.8,5.4Hz,1H),2.64–2.58(m,1H),2.56–2.50(m,5H),2.10–2.03(m,1H). 13C NMR (151MHz, DMSO-d6) δ172.66, 169.51, 162.67, 153.05 (d, J = 246.6Hz), 145.07, 143.75, 137.65, 135.16, 129.80, 128.93, 127. 84,121.69,119.67,119.50,119.31,118.46,118.30,114.71,112.86,102.11,69.37,61.49,53.75,52.13,49.13,30.82,22.24.

[0212] Example 3: (S)-4-(4-((5-(((2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazol-7-yl)oxy)methyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile (D3)

[0213] Synthesis route:

[0214] Step 1: Dissolve N-Boc-4-mercaptopiperidine (1.05 g, 4.83 mmol), 2-fluoropyridine-5-carboxaldehyde (664 mg, 5.31 mmol) and potassium carbonate (1.67 g, 12.08 mmol) in N, N-dimethylformamide (10 mL). Heat the reaction solution to 80°C and stir for 1 hour. Remove the solvent from the reaction solution under reduced pressure, add water to dilute, extract with ethyl acetate, combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, remove the solvent under reduced pressure, and separate the residue by silica gel column chromatography to obtain 1.5 g of the product with a yield of 96%. UPLC-MS (ESI) theoretical value is C 16 H 22 N2O3S[M+H] + :323.42, measured value: 323.53.

[0215] 1 H NMR (500MHz, DMSO-d6) δ10.00(s,1H),8.92(dd,J=2.2,0.8Hz,1H),8.02(dd,J=8.4,2.3Hz,1H),7.48(d,J=8.4Hz,1H),4.10( tt,J=10.4,3.9Hz,1H),3.85–3.77(m,2H),3.08(s,2H),2.07–1.98(m,2H),1.53(dtd,J=13.1,10.5,4.1Hz,2H),1.41(s,9H).

[0216] Step 2: Dissolve the compound tert-butyl 4-((5-formylpyridin-2-yl)thio)piperidine-1-carboxylate (1.5 g) in dichloromethane (10 mL), add trifluoroacetic acid (3 mL) dropwise under ice-cooling, and react for 1 hour. The reaction solution is concentrated and dried to obtain 1.5 g of the product. The crude product is used directly in the next reaction without purification. The theoretical value of UPLC-MS (ESI) is C 11 H 14 N2OS[M+H] + :223.31, measured value: 223.14

[0217] Step 3: Dissolve the compound 6-(piperidin-4-thio)nicotinaldehyde trifluoroacetate (1.5 g, 4.46 mmol), 3,4-difluorobenzonitrile (1.24 g, 8.92 mmol) and potassium carbonate (1.85 g, 13.38 mmol) in N,N-dimethylformamide (15 mL). Stir the reaction solution at 80°C overnight. Concentrate the reaction solution under reduced pressure, dilute with water, extract with ethyl acetate, combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The resulting residue is subjected to column chromatography to obtain 1.33 g of the product with a yield of 87%. The theoretical value of UPLC-MS (ESI) is C 18 H 16 FN3OS[M+H] + :342.40, measured value: 342.29. 1 H NMR(500MHz,Chloroform-d)δ10.00(s,1H),8.83(dd,J=2.2,0.9Hz,1H),7.93(d d,J=8.4,2.2Hz,1H),7.37(ddd,J=8.4,1.9,0.8Hz,1H),7.30–7.27(m,2H),6.95 (t,J=8.5Hz,1H),4.21(tt,J=10.1,4.1Hz,1H),3.60–3.54(m,2H),3.11(ddd,J= 12.7,10.1,2.9Hz,2H),2.32–2.22(m,2H),1.95(dtd,J=13.7,10.1,3.7Hz,2H). 13 C NMR (126MHz, Chloroform-d) δ 189.95, 166.26, 154.00 (d, J = 248.6Hz), 152.58, 144.28, 134.62, 129.42 (d, J = 3. 4Hz), 128.03, 122.59, 119.74 (d, J = 25.0Hz), 119.01 (d, J = 3.8Hz), 118.44, 103.61, 49.71, 49.68, 40.10, 31.91.

[0218] Step 4: Add 3-fluoro-4-(4-((5-formylpyridin-2-yl)thio)piperidin-1-yl)benzonitrile (1.33 g, 3.90 mmol) to tetrahydrofuran (20 mL) under ice-bath conditions, slowly add sodium borohydride (294 mg, 7.79 mmol), and react for 1 hour. The reaction solution is quenched with water, then extracted with ethyl acetate, the combined organic phases are washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate is concentrated under reduced pressure. The resulting residue is subjected to column chromatography to obtain 1.24 g of the product in a yield of 93%. The theoretical value of UPLC-MS (ESI) is C 18 H 18 FN3OS[M+H] + :344.42, measured value: 344.30. 1 H NMR(500MHz,Chloroform-d)δ8.41(d,J=2.2Hz,1H),7.54(dd,J=8.2,2.3Hz,1H),7.3 5(dd,J=8.5,1.9Hz,1H),7.29–7.23(m,1H),7.19(d,J=8.2Hz,1H),6.93(t,J=8.5Hz, 1H),4.66(s,2H),4.09–4.02(m,1H),3.55(dt,J=11.8,4.3Hz,2H),3.07(ddd,J=12.7 ,9.9,2.9Hz,2H),2.23(dq,J=12.8,3.9Hz,2H),1.90(dtd,J=13.5,10.0,3.6Hz,2H).

[0219] Step 5: Compound 3-fluoro-4-(4-((5-(hydroxymethyl)pyridin-2-yl)thio)piperidin-1-yl)benzonitrile (376 mg, 1.1 mmol), methyl 2-nitro-3-hydroxybenzoate (180 mg, 0.91 mmol) and triphenylphosphine (287 mg, 1.1 mmol) were added to dry tetrahydrofuran (5 mL). Diisopropyl azodicarboxylate (221 mg, 1.1 mmol) was slowly added dropwise at 0°C. The mixture was stirred overnight and the reaction solution was concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain 458 mg of the product in a yield of 96%. The theoretical value of UPLC-MS (ESI) was C 26 H 23 FN4O5S[M+H] + :523.55, measured value: 523.33. 1H NMR(500MHz,Chloroform-d)δ8.45–8.41(m,1H),7.64(dd,J=7.9,1.1Hz,1H),7.53(dd,J=8.3,2.3Hz,1H ),7.50(t,J=8.2Hz,1H),7.35(ddd,J=8.5,2.1,0.8Hz,1H),7.31(dd,J=8.4,1.2Hz,1H),7.27(dd,J=12.6 ,2.0Hz,1H),7.20(dd,J=8.2,0.9Hz,1H),5.14(s,2H),4.08(tt,J=9.9,4.0Hz,1H),3.91(s,3H),3.59–3 .52(m,2H),3.09(ddd,J=12.7,10.1,2.9Hz,2H),2.28–2.21(m,2H),1.91(dtd,J=13.6,10.0,3.7Hz,2H).

[0220] Step 6: Add the compound 3-((6-((1-(4-cyano-2-fluorophenyl)piperidin-4-yl)sulfanyl)pyridin-3-yl)methoxy)-2-nitrobenzoic acid methyl ester (458 mg, 0.87 mmol) to tetrahydrofuran (10 mL), add an aqueous solution (3 mL) of lithium hydroxide monohydrate (37 mg, 0.87 mmol), stir at room temperature overnight, concentrate the reaction solution under reduced pressure, and dry to obtain 450 mg of the product, which is used directly in the next reaction without purification.

[0221] Step 7: Compound 3-((6-((1-(4-cyano-2-fluorophenyl)piperidin-4-yl)thio)pyridin-3-yl)methoxy)-2-nitrobenzoic acid lithium salt (450 mg, 0.87 mmol), (S)-tert-butyl 4,5-diamino-5-oxopentanoate monohydrochloride (208 mg, 0.87 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (332 mg, 0.87 mmol) were added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.457 mL, 2.62 mmol) was added dropwise. The reaction mixture was reacted at room temperature for 12 hours. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain 550 mg of the product in a yield of 91%. The theoretical value of UPLC-MS (ESI) is C 34 H 37 FN6O7S[M+H] + :693.76, measured value: 693.43. 1H NMR(500MHz,Chloroform-d)δ8.44(d,J=2.3Hz,1H),7.57–7.44(m,3H),7.35(dd,J=8.5,1.9Hz,1H),7.26–7.17(m,3 H),6.94(t,J=8.5Hz,1H),5.14(s,2H),4.59(td,J=8.1,7.7,4.7Hz,1H),4.07(dq,J=9.5,4.8,4.2Hz,1H),3.72(tt,J =6.3,3.0Hz,1H),3.55(dt,J=12.6,4.3Hz,2H),3.17(dt,J=11.8,5.7Hz,1H),3.12–3.05(m,2H),2.67–2.51(m,1H), 2.40(dt,J=16.9,6.5Hz,1H),2.29–2.13(m,3H),2.04(q,J=7.5Hz,1H),1.95–1.86(m,2H),1.79(s,1H),1.44(s,9H).

[0222] Step 8: Add the compound 5-amino-4-(3-((6-((1-(4-cyano-2-fluorophenyl)piperidin-4-yl)thio)pyridin-3-yl)methoxy)-2-nitrobenzamido)-5-oxopentanoic acid tert-butyl ester (550 mg, 0.79 mmol) to methanol (7 mL), add tetrahydroxydiboron (355 mg, 3.97 mmol) at 0°C, and then add a methanol solution (8 mL) of sodium hydroxide (317 mg, 7.94 mmol). The reaction was carried out at 0°C for 10 minutes, and then the temperature was raised to 40°C for overnight reaction. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain 443 mg of the product with a yield of 84%. The theoretical value of UPLC-MS (ESI) was C 34 H 37 FN6O5S[M+H] + :661.77, measured value: 661.46.

[0223] Step 9: Add tert-butyl 5-amino-4-(7-((6-((1-(4-cyano-2-fluorophenyl)piperidin-4-yl)thio)pyridin-3-yl)methoxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)-5-oxopentanoate (150 mg, 0.23 mmol) and p-toluenesulfonic acid monohydrate (259 mg, 1.36 mmol) to acetonitrile (3 mL) and react at 80°C overnight. The reaction solution was concentrated and purified by HPLC to obtain 35 mg of the product with a yield of 26%. The theoretical value of UPLC-MS (ESI) was C 30H 27 FN6O4S[M+H] + :587.64, measured value: 587.33. 1 H NMR (500MHz, DMSO-d6) δ10.99(s,1H),10.08(s,1H),8.65(d,J=2.2Hz,1H),7.81(dd,J=8.3,2.3Hz,1H),7.69(dd,J=13.3,2.0Hz,1H),7.56(dd, J=8.4,2.0Hz,1H),7.36(d,J=8.2Hz,1H),7.27(d,J=7.8Hz,1H),7.23(d,J=7.8Hz,1H),7.16(t,J=8.8Hz,1H),7.08(t,J=7.8Hz,1H),5.32(dd,J =13.1,5.3Hz,1H),5.24(s,2H),4.03(tt,J=10.2,4.0Hz,1H),3.52(dt,J=12.8,4.1Hz,2H),3.08(ddd,J=12.9,10.3,2.7Hz,2H),2.93(ddd,J=1 7.2,13.7,5.4Hz,1H),2.65–2.51(m,2H),2.16(dq,J=12.1,3.6Hz,2H), 2.07(dtd,J=12.9,5.4,2.4Hz,1H), 1.76(dtd,J=13.7,10.3,3.6Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ172.76,169.62,162.75,157.46,153.14(d,J=246.1Hz),149.18,144.89,143.98,137.75,136.68,129.90,128.28,122 .09,121.80,119.82,119.67,119.62,118.58,118.44,115.07,113.03, 102.07(d,J=9.9Hz),66.99,53.87,49.20,49.16,31.71,30.93,22.37.

[0224] Example 4: 4-(4-((5-(((2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazol-7-yl)oxy)methyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile (D4)

[0225] The synthesis method was similar to that of Example 1 to obtain compound D4. 1H NMR (500MHz, DMSO-d6) δ11.00(s,1H),10.14(s,1H),7.74(dd,J=13.2,2.0Hz,1H),7.63(d,J=7.9Hz,2 H),7.60(dd,J=8.4,1.9Hz,1H),7.52–7.46(m,2H),7.27(d,J=7.8Hz,1H),7.21(d,J=7.8Hz,1H),7.16( t, J = 8.7 Hz, 1H), 7.07 (t, J = 7.8 Hz, 1H), 5.36–5.29 (m, 3H), 3.78 (s, 2H), 3.57–3.40 (m, 2H), 3.29–3.14 (m, 4H), 2.93 (ddd, J = 17.2, 13.8, 5.5 Hz, 1H), 2.66–2.54 (m, 2H), 2.12–2.00 (m, 1H). UPLC-MS (ESI) theoretical value was C 31 H 28 FN6O5[M+H] + :Calculated value 583.21, measured value 583.38.

[0226] Example 5: 3-(7-((4-(4-(2-fluoro-4-methylphenyl)piperazine-1-carbonyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D5)

[0227] The synthesis method was similar to that of Example 1 to obtain compound D5. 1 H NMR (500 MHz, DMSO-d6) δ 11.00 (s, 1H), 10.14 (s, 1H), 7.62 (d, J = 7.9 Hz, 2H), 7.48 (d, J = 8.1 Hz, 2H), 7.27 (d, J = 7.8 Hz, 1H), 7.21 (d, J = 7.8 Hz, 1H), 7.07 (t, J = 7.8 Hz, 1H), 7.00–6.89 (m, 3H), 5.35–5.28 (m, 3H), 3.77 (s, 2H), 3.49 (s, 2H), 3.07–2.86 (m, 5H), 2.65–2.53 (m, 2H), 2.24 (s, 3H), 2.11–2.04 (m, 1H). UPLC-MS (ESI) requires C 31 H 31 FN5O5[M+H] + :Calculated value 572.23, measured value 572.32.

[0228] Example 6: 3-(7-((4-(((2S,6R)-2,6-dimethylmorpholino)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D6)

[0229] The synthesis method was similar to that in Example 1 to obtain compound D6. 1 H NMR (500MHz, DMSO-d6) δ11.00(s,1H),10.11(s,1H),7.65(d,J=7.9Hz,2H),7.56(d,J=8.0Hz,2H),7.28( d,J=7.7Hz,1H),7.21(d,J=7.8Hz,1H),7.07(t,J=7.8Hz,1H),5.39–5.27(m,3H),4.34(s,2H),3.82(tdd, J=12.4,6.1,1.9Hz,3H),3.27(s,1H),2.94(ddd,J=17.1,13.8,5.4Hz,1H),2.69(t,J=11.5Hz,2H),2.63(dt,J=17.7,3.2Hz,1H),2.58–2.52(m,1H),2.13–2.03(m,1H),1.12(d,J=6.3Hz,6H). UPLC-MS (ESI) theoretical value is C 26 H 31 N4O5[M+H] + :Calculated value 479.22, measured value 479.45.

[0230] Example 7: 3-(7-((4-((4-(2,4-difluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D7)

[0231] The synthesis method was similar to that of Example 1 to obtain compound D7. 1H NMR (500MHz, DMSO) δ10.98(s,1H),10.11(s,1H),7.50(d,J=7.6Hz,2H),7.37(d,J=7.6Hz,2 H),7.25(d,J=7.8Hz,1H),7.23–7.15(m,2H),7.06(td,J=8.7,5.7Hz,2H),6.99(t,J=5.3Hz, 1H), 5.31 (dd, J = 13.1, 5.1 Hz, 1H), 5.23 (s, 2H), 3.54 (s, 2H), 3.35 (s, 4H), 2.96 (s, 4H), 2.93–2.87 (m, 1H), 2.66–2.57 (m, 1H), 2.54–2.51 (m, 1H), 2.11–2.02 (m, 1H). UPLC-MS (ESI) theoretical value was C 30 H 29 F2N5O4[M+H] + :Calculated value 562.22, measured value 562.37.

[0232] Example 8: 3-(7-((4-((4-(3-chlorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D8)

[0233] The synthesis method was similar to that in Example 1 to obtain compound D8. 1 H NMR (500MHz, DMSO) δ10.98(s,1H),10.10(s,1H),7.51(d,J=7.5Hz,2H),7.38(d,J=7.7Hz,2H),7.26 (d,J=7.9Hz,1H),7.21(d,J=8.0Hz,2H),7.07(t,J=7.8Hz,1H),6.92(s,1H),6.88(d,J=8.6Hz,1H),6 .78(d,J=7.8Hz,1H),5.31(dd,J=13.1,5.1Hz,1H),5.24(s,2H),3.54(s,2H),3.34-3.29(m,4H),3.17(s,4H),2.98–2.87(m,1H),2.63(d,1H),2.54-2.51(m,1H),2.10–2.04(m,1H). UPLC-MS(ESI)theoretical value is C 30 H 31 ClN5O4[M+H] + :Calculated value 560.20, measured value 560.28.

[0234] Example 9: 3-(7-((4-((4-(4-chlorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D9)

[0235] The synthesis method was similar to that of Example 1 to obtain compound D9. 1 H NMR(500MHz,DMSO)δ10.98(s,1H),10.10(s,1H),7.51(d,J=7.7Hz,2H),7.37(d,J=7.6Hz,2H), 7.26(d,J=7.8Hz,1H),7.23–7.19(m,3H),7.07(t,J=7.8Hz,1H),6.96–6.90(m,2H),5.31(dd,J =13.1,5.2Hz,1H),5.23(s,2H),3.56–3.52(m,2H),3.34-3.29(m,4H),3.13(t,J=5.0Hz,4H),2.98–2.87(m,1H),2.66–2.58(m,1H),2.54-2.51(m,1H),2.10–2.04(m,1H). UPLC-MS (ESI) theoretical value was C 30 H 31 ClN5O4[M+H] + :Calculated value 560.20, measured value 560.32.

[0236] Example 10: 3-(7-((4-((4-(4-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D10)

[0237] The synthesis method was similar to that of Example 1 to obtain compound D10. 1H NMR (500MHz, DMSO) δ10.98(s,1H),10.10(s,1H),7.51(d,J=7.7Hz,2H),7.38(d,J=7.7Hz,2H),7.2 6(d,J=7.8Hz,1H),7.21(d,J=7.8Hz,1H),7.08(d,J=7.8Hz,1H),7.06–7.01(m,2H),6.95–6.91(m,2 H), 5.31 (dd, J = 13.1, 5.2 Hz, 1H), 5.23 (s, 2H), 3.54 (d, J = 2.6 Hz, 2H), 3.34-3.29 (m, 4H), 3.07 (s, 4H), 2.98–2.87 (m, 1H), 2.64–2.57 (m, 1H), 2.55–2.52 (m, 1H), 2.10–2.04 (m, 1H). UPLC-MS (ESI) theoretical value is C 30 H 31 FN5O4[M+H] + :Calculated value 544.23, measured value 544.32.

[0238] Example 11: 3-(3-oxo-7-((4-((4-(4-(trifluoromethyl)phenyl)piperazin-1-yl)methyl)benzyl)oxy)-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D11)

[0239] The synthesis method was similar to that of Example 1 to obtain compound D11. 1 H NMR (500MHz, DMSO) δ11.00(s,1H),10.18(s,1H),7.63(q,J=8.2Hz,2H),7.55(d,J=8.4Hz,1H ),7.42(s,2H),7.32(s,2H),7.26(d,J=8.1Hz,1H),7.22(d,J=7.1Hz,2H),7.09(dd,J=13.6,8 .1Hz,1H),5.32(dd,J=12.9,5.1Hz,1H),5.28(s,2H),4.22(s,2H),3.57(s,2H),3.51(s,2H),3.49(s,3H),3.47(s,2H),2.89(m,1H),2.67–2.55(m,1H),2.08(m,1H). UPLC-MS (ESI) theoretical value is C 30 H 31 F3N5O4[M+H] + :Calculated value 594.22, measured value 594.34.

[0240] Example 12: 3-(7-((4-((4-(2,4-dichlorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D12)

[0241] The synthesis method was similar to that of Example 1 to obtain compound D12. 1 H NMR(500MHz,DMSO)δ11.01(s,1H),10.12(s,1H),7.70–7.64(m,2H),7.64–7. 56(m,3H),7.42(dd,J=8.7,2.5Hz,1H),7.30–7.22(m,2H),7.21(d,J=7.8Hz,1 H),7.07(t,J=7.8Hz,1H),5.34(d,J=5.2Hz,1H),5.31(s,2H),4.45(s,2H),3 .57(t,J=5.5Hz,4H),3.25(s,2H),3.08–2.97(m,2H),2.97–2.88(m,1H),2.62 (m, 1H), 2.54-2.51 (m, 1H), 2.09 (m, 1H). UPLC-MS (ESI) theoretical value is C 30 H 30 Cl2N5O4[M+H] + :Calculated value 594.16, measured value 594.25.

[0242] Example 13: 3-(7-((4-((3-morpholinoazetidin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D13)

[0243] The synthesis method was similar to that of Example 1 to obtain compound D13. 1H NMR(500MHz,DMSO)δ11.00(s,1H),7.62(d,J=8.0Hz,2H),7.52(d,J=8.0Hz,2H),7.27 (d,J=7.8Hz,1H),7.20(d,J=7.7Hz,1H),7.07(t,J=7.8Hz,1H),5.32(dd,J=13.2,5.2 Hz,1H),5.29(s,2H),4.45(s,2H),4.20(s,4H),3.73(s,5H),3.42(s,1H),2.99–2.88 (m,1H),2.78–2.68(m,3H),2.65–2.60(m,1H),2.54-2.51(m,1H),2.13–2.05(m,1H).

[0244] The theoretical value of UPLC-MS (ESI) is C 27 H 32 N5O5[M+H] + :Calculated value 506.23, measured value 506.34.

[0245] Example 14: 4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-5-fluoro-3-oxo-2,3-dihydro-1H-indazol-7-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile (D14)

[0246] The synthesis method was similar to that of Example 1 to obtain compound D14. 1 H NMR (400MHz, DMSO) δ11.00(s,1H),10.11(s,1H),7.78(dd,J=13.1,1.9Hz,1H),7.67–7.61(m,3H), 7.57(d,J=7.9Hz,2H),7.22(dd,J=10.3,7.2Hz,2H),7.04(dd,J=7.6,2.2Hz,1H),5.31(d,J=12.8H z, 3H), 4.42(s, 2H), 3.72(d, J=13.0Hz, 2H), 3.41(s, 2H), 3.25(s, 2H), 3.16(d, J=12.6Hz, 2H), 2.98–2.90(m, 1H), 2.61(d, J=16.8Hz, 1H), 2.54–2.51(m, 1H), 2.13–2.03(m, 1H). UPLC-MS (ESI) theoretical value is C 31 H 29 F2N6O4[M+H] + :Calculated value 587.21, measured value 587.62.

[0247] Example 15: 3-(7-((4-((4-(2,4-difluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-5-fluoro-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D15)

[0248] The synthesis method was similar to that of Example 1 to obtain compound D15. 1 H NMR (500MHz, DMSO) δ11.02 (s, 1H), 10.14 (s, 1H), 7.66 (d, J = 7.7Hz, 2H), 7.59 ( d,J=7.8Hz,2H),7.32–7.20(m,2H),7.15(td,J=9.5,5.8Hz,1H),7.05(dd,J=7 .5,2.5Hz,2H),5.34(s,3H),4.44(s,2H),4.2(s,2H),3.44(s,4H),3.26(s,2H ),2.95–2.89(m,1H),2.66–2.58(m,1H),2.54–2.51(m,1H),2.14–2.06(m,1H).

[0249] The theoretical value of UPLC-MS (ESI) is C 31 H 29 F2N6O4[M+H] + :Calculated value 580.21, measured value 580.63.

[0250] Example 16: 3-(7-((4-((4-(2,4-dichlorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-5-fluoro-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D16)

[0251] The synthesis method was similar to that of Example 1 to obtain compound D16. 1H NMR(500MHz,DMSO)δ11.00(s,1H),10.15(s,1H),7.56–7.48(m,3H),7.41–7.36(m,3 H),7.26–7.20(m,1H),7.17(d,J=8.7Hz,1H),7.03(dd,J=7.5,2.3Hz,1H),5.32(dd,J =13.0,5.1Hz,1H),5.25(s,2H),3.57(s,2H),3.35(s,4H),2.97(s,4H),2.93–2.88(m,1H),2.62–2.58(m,1H),2.54–2.51(m,1H),2.10–2.04(m,1H). UPLC-MS (ESI) theoretical value was C 30 H 29 C l2 FN5O4[M+H]+: calcd. 612.15, found 612.30.

[0252] Example 17: 3-(5-Fluoro-3-oxo-7-((4-((4-(4-(trifluoromethyl)phenyl)piperazin-1-yl)methyl)benzyl)oxy)-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D17)

[0253] The synthesis method was similar to that of Example 1 to obtain compound D17. 1 H NMR (500MHz, DMSO) δ11.01(s,1H),10.15(s,1H),7.51(t,J=8.0Hz,4H),7.40(d,J=7.7H z,2H),7.23(dd,J=11.3,2.3Hz,1H),7.05(dd,J=8.1,5.6Hz,3H),5.32(dd,J=13.2,5.2H z, 1H), 5.26 (s, 2H), 3.55 (d, J = 8.5 Hz, 2H), 3.35 (s, 4H), 3.28 (t, J = 5.0 Hz, 4H), 2.98–2.87 (m, 1H), 2.65–2.57 (m, 1H), 2.54–2.51 (m, 1H), 2.12–2.03 (m, 1H). UPLC-MS (ESI) theoretical value is C 31 H 30 F4N5O4[M+H]+: calcd. 612.22, found 612.36.

[0254] Example 18: 3-(7-((4-((4-(4-chlorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-5-fluoro-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D18)

[0255] The synthesis method was similar to that of Example 1 to obtain compound D18. 1 H NMR (500MHz, DMSO) δ11.01(s,1H),10.15(s,1H),7.51(d,J=7.7Hz,2H),7.39(d,J=7.7Hz,2H ),7.23(dd,J=9.5,2.9Hz,3H),7.04(dd,J=7.6,2.2Hz,1H),6.96–6.90(m,2H),5.32(dd,J=1 3.1,5.2Hz,1H),5.25(s,2H),3.54(d,J=4.3Hz,2H),3.35(s,4H),3.13(t,J=4.9Hz,4H),2.98–2.87(m,1H),2.65–2.58(m,1H),2.54–2.51(m,1H),2.08–2.05(m,1H). UPLC-MS (ESI) theoretical value was C 30 H 30 ClFN5O4[M+H] + :Calculated value 578.19, measured value 578.36.

[0256] Example 19: 3-(5-Fluoro-7-((4-((4-(4-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D19)

[0257] The synthesis method was similar to that of Example 1 to obtain compound D19. 1H NMR (500MHz, DMSO) δ11.01(s,1H),10.15(s,1H),7.51(d,J=7.7Hz,2H),7.39(d,J=7.7Hz,2H),7.23(dd,J=1 1.2,2.2Hz,1H),7.08–7.00(m,3H),6.97–6.89(m,2H),5.32(dd,J=13.2,5.2Hz,1H),5.25(s,2H),3.54(d,J= 6.3Hz,2H),3.35(s,4H),3.07(d,J=5.8Hz,4H),2.97–2.87(m,1H),2.64–2.57(m,1H),2.54–2.51(m,1H),2.11–2.03(m,1H). UPLC-MS (ESI) theoretical value is C 30 H 30 F2N5O4[M+H] + :Calculated value 562.22, measured value 562.33.

[0258] Example 20: 3-(5-fluoro-7-((4-((3-morpholinoazetidin-1-yl)methyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D20)

[0259] The synthesis method was similar to that of Example 1 to obtain compound D20. 1 H NMR (500MHz, DMSO) δ11.01(s,1H),10.15(s,1H),7.48(d,J=7.8Hz,2H),7.34(d,J=8.0H z,2H),7.21(dd,J=11.2,2.3Hz,1H),7.03(dd,J=7.6,2.2Hz,1H),5.32(dd,J=13.2,5.2H z,1H),5.24(s,2H),3.67(s,2H),3.60–3.51(m,5H),3.42(s,1H),2.98–2.86(m,4H),2.65–2.56(m,1H),2.54–2.51(m,1H),2.23(s,4H),2.11–2.03(m,1H). UPLC-MS (ESI) theoretical value is C 27 H 31 FN5O5[M+H] + :Calculated value 524.22, measured value 524.47.

[0260] Example 21: 3-(7-((4-((4-(4-chloro-2-fluorophenyl)piperazin-1-yl)methyl)benzyl)oxy)-5-fluoro-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D21)

[0261] The synthesis method was similar to that of Example 1 to obtain compound D21. 1 H NMR (500MHz, DMSO) δ11.02(s,1H),10.17(s,1H),7.66(d,J=8.0Hz,2H),7.59(d,J=8.1Hz,2H),7 .42(dd,J=12.2,2.4Hz,1H),7.26–7.20(m,2H),7.12(t,J=9.1Hz,1H),7.05(dd,J=7.6,2.2Hz,1 H), 5.37–5.30(m,3H),4.45(s,2H),3.55–3.48(m,2H),3.42(s,2H),3.25(s,2H),3.05(s,2H),2.99–2.88(m,1H),2.66–2.58(m,1H),2.54–2.51(m,1H),2.14–2.05(m,1H). UPLC-MS (ESI) theoretical value is C 30 H 29 ClF2N5O4[M+H] + :596.18, measured value 596.27.

[0262] Example 22: 3-(1-methyl-7-((4-morpholinomethyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D22)

[0263] The synthesis method was similar to that of Example 1 to obtain compound D22. 1H NMR (500MHz, DMSO) δ11.03(s,1H),7.61(d,J=7.7Hz,2H),7.57(d,J=6.9Hz,2H),7.33(d, J=8.0Hz,1H),7.29(d,J=7.8Hz,1H),7.20(dd,J=8.8,6.8Hz,1H),5.35(s,2H),5.10–5.0 4 (m, 1H), 4.37 (s, 2H), 3.97 (d, J = 12.9 Hz, 4H), 3.65 (d, J = 12.3 Hz, 4H), 2.81 (t, J = 14.6 Hz, 1H), 2.72–2.58 (m, 2H), 2.04 (d, J = 11.9 Hz, 1H), 1.27–1.19 (m, 3H). UPLC-MS (ESI) theoretical value was C 25 H 29 N4O5[M+H] + :465.52, measured value 465.37.

[0264] Example 23: 3-(1-ethyl-7-((4-morpholinomethyl)benzyl)oxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D23)

[0265] The synthesis method was similar to that of Example 1 to obtain compound D23. 1 H NMR (500MHz, DMSO) δ11.03(s,1H),7.61(d,J=8.0Hz,2H),7.57(d,J=7.9Hz,2H),7.33(d,J=7.9Hz,1 H),7.28(d,J=7.6Hz,1H),7.20(t,J=7.8Hz,1H),5.35(s,2H),5.07(dd,J=12.6,5.3Hz,1H),4.37(s ,2H),4.11(dq,J=13.8,6.7Hz,2H),3.97(d,J=12.9Hz,4H),3.64(t,J=12.2Hz,4H),3.27(d,J=12.8Hz,2H),2.87–2.76(m,1H),2.72–2.57(m,2H),2.07–1.99(m,1H),1.23(s,3H). UPLC-MS (ESI) theoretical value was C 26 H 31 N4O5[M+H] + :479.55, measured value 479.86.

[0266] Example 24: 4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-methyl-3-oxo-2,3-dihydro-1H-indazol-7-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile (D24)

[0267] The synthesis method was similar to that of Example 1 to obtain compound D24. 1 H NMR (500MHz, DMSO) δ11.05(s,1H),7.83–7.76(m,1H),7.64(d,J=10.5Hz,3H),7.59(d,J=7.8Hz,2H ),7.34(dd,J=7.9,1.0Hz,1H),7.30(dd,J=7.8,0.9Hz,1H),7.22(td,J=8.2,2.7Hz,2H),5.35(s,2 H), 5.14 (dd, J = 12.6, 5.3 Hz, 1H), 4.44 (s, 2H), 3.73 (d, J = 13.1 Hz, 3H), 3.26 (s, 3H), 3.19 (d, J = 12.6 Hz, 2H), 2.87–2.78 (m, 1H), 2.73–2.58 (m, 2H), 2.10–2.04 (m, 1H), 1.24 (s, 3H) UPLC-MS (ESI) theoretical value is C 32 H 32 FN6O4[M+H] + :583.64, measured value 583.49.

[0268] Example 25: 4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-methyl-3-oxo-2,3-dihydro-1H-indazol-7-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile (D25)

[0269] The synthesis method was similar to that of Example 1 to obtain compound D25. 1H NMR(500MHz,DMSO)δ11.04(s,1H),7.79(dd,J=13.1,1.9Hz,1H),7.64–7.58(m,5H),7.36– 7.27(m,2H),7.23–7.17(m,2H),5.35(s,2H),5.07(dd,J=12.5,5.3Hz,1H),4.43(s,2H),4 .12 (dq, J = 14.0, 6.9 Hz, 1H), 3.79 (dd, J = 15.0, 7.1 Hz, 2H), 3.44 (d, J = 12.2 Hz, 4H), 2.87–2.76 (m, 1H), 2.71–2.58 (m, 2H), 2.07–2.01 (m, 1H), 1.30–1.22 (m, 3H). UPLC-MS (ESI) theoretical value was C 33 H 34 FN6O4[M+H] + :597.66, measured value 597.35.

[0270] Example 26: (S)-3-(3-oxo-7-((2-(4-(trifluoromethoxy)phenyl)oxazol-5-yl)methoxy)-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D26)

[0271] Synthesis route:

[0272] Step 1:

[0273] The compound 4-trifluoromethoxyphenylboronic acid (1g, 4.89mmol), ethyl 2-iodooxazole-5-carboxylate (1g, 3.25mmol), sodium carbonate (795mg, 7.5mmol) and tetrakistriphenylphosphine palladium (433mg, 0.375mmol) were added to a mixed solvent of 1,4-dioxane (15mL) and water (3mL). The reaction system was replaced with nitrogen and heated to 90°C under nitrogen protection to react overnight. 1,4-dioxane was removed by concentration under reduced pressure, and the residue was extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was separated by silica gel column chromatography to obtain 661mg of the product as a yellow oil with a yield of 59%. The theoretical value of UPLC-MS (ESI) was C 13 H 10 F3NO4[M+H] + :302.22, measured value: 302.17.

[0274] Step 2:

[0275] The compound ethyl 2-(4-(trifluoromethoxy)phenyl)oxazole-5-carboxylate (611 mg, 2 mmol) was dissolved in ethanol (15 mL), and calcium chloride (1.3 g, 12 mmol) was added. The reaction system was cooled to 0°C, and sodium borohydride (227 mg, 6 mmol) was slowly added. The temperature was raised to room temperature and the reaction was allowed to react overnight. Saturated ammonium chloride solution (5 mL) was added to the reaction system to quench the reaction, diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was separated by silica gel column chromatography to obtain 311 mg of the product as a white solid with a yield of 59%. The theoretical value of UPLC-MS (ESI) was C 11 H8F3NO3[M+H] + :260.18, measured value: 260.26.

[0276] Step 3:

[0277] The compound (2-(4-(trifluoromethoxy)phenyl)oxazol-5-yl)methanol (200 mg, 1 mmol), methyl 2-nitro-3-hydroxybenzoate (315 mg, 1.2 mmol) and triphenylphosphine (315 mg, 1.2 mmol) were added to dry tetrahydrofuran (8 mL). Diisopropyl azodicarboxylate (0.236 mL) was slowly added dropwise at 0°C and stirred overnight. The reaction solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 400 mg of the product as a yellow oil with a yield of 89%. The theoretical value of UPLC-MS (ESI) was C 19 H 13 F3N2O7[M+H] + :439.32, measured value: 439.40.

[0278] Step 4:

[0279] The compound methyl 2-nitro-3-((2-(4-(trifluoromethoxy)phenyl)oxazol-5-yl)methoxy)benzoate (440 mg, 1 mmol) was added to tetrahydrofuran (8 mL), and an aqueous solution (4 mL) of lithium hydroxide monohydrate (63 mg, 1.5 mmol) was added. The reaction was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure and dried to obtain 430 mg of the product, which was used directly in the next reaction without purification.

[0280] Step 5:

[0281] The compound 2-nitro-3-((2-(4-(trifluoromethoxy)phenyl)oxazol-5-yl)methoxy)benzoic acid lithium salt (431 mg, 1 mole), (S)-4,5-diamino-5-oxopentanoic acid tert-butyl ester monohydrochloride (239 mg, 1 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (381 mg, 1 mmol) were added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.523 mL, 3 mmol) was added dropwise. The reaction was allowed to react at room temperature for 12 hours. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 530 mg of the product with a yield of 87%. The theoretical value of UPLC-MS (ESI) was C 27 H 27 F3N4O9[M+H] + :609.53, measured value: 609.50.

[0282] Step 6:

[0283] Compound (S)-tert-butyl 5-amino-4-(2-nitro-3-((2-(4-(trifluoromethoxy)phenyl)oxazol-5-yl)methoxy)benzamido)-5-oxopentanoate (420 mg, 0.69 mmol) was added to methanol (7 mL). Tetrahydroxydiboron (309 mg, 3.45 mmol) was added at 0°C, followed by a methanol solution (9 mL) of sodium hydroxide (276 mg, 6.9 mmol). The mixture was allowed to react at 0°C for 10 minutes, then raised to 40°C and allowed to react overnight. After TLC monitoring, the reaction was quenched by adding saturated ammonium chloride solution, extracted with ethyl acetate, and separated. The organic phase was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 123 mg of the product as a yellow oil with a yield of 31%. The theoretical value of UPLC-MS (ESI) was C 27 H 27 F3N4O7[M+H] + :577.53, the measured value is 577.56:

[0284] Step 7:

[0285] Compound (S)-tert-butyl 5-amino-5-oxo-4-(3-oxo-7-((2-(4-(trifluoromethoxy)phenyl)oxazol-5-yl)methoxy)-1,3-dihydro-2H-indazol-2-yl)pentanoate (123 mg, 0.21 mmol) and p-toluenesulfonic acid monohydrate (240 mg, 1.26 mol) were added to acetonitrile (6 mL) and reacted at 80°C overnight. The reaction solution was concentrated and purified by HPLC to obtain 35 mg of the product in a yield of 33%. The theoretical value of UPLC-MS (ESI) was C 23 H 17 F3N4O6[M+H] + :503.41, measured value:503.31. 1 H NMR(500MHz,DMSO-d6)δ11.04(s,1H),10.72(s,1H),7.76(d,J=8.9Hz,2H),7 .47(d,J=8.2Hz,2H),7.14–7.11(m,1H),7.08(t,J=7.6Hz,1H),7.05(s,1H), 7.03–7.00(m,1H),5.37(d,J=16.3Hz,1H),5.17(dd,J=12.4,5.1Hz,1H),5.0 9(d,J=16.4Hz,1H),2.91–2.81(m,1H),2.76–2.64(m,2H),2.28–2.21(m,1H).

[0286] Example 27: (S)-3-(7-((2-(4-chlorophenyl)oxazol-5-yl)methoxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D27)

[0287] The synthesis method was similar to that of Example 26 to obtain compound D27. 1 H NMR (500 MHz, DMSO) δ 11.02 (s, 1H), 10.59 (s, 1H), 7.61–7.55 (m, 2H), 7.15–7.05 (m, 3H), 7.05–6.97 (m, 3H), 5.40–5.31 (m, 2H), 5.17 (dd, J = 12.5, 5.2 Hz, 1H), 2.71 (dd, J = 17.2, 4.5 Hz, 2H), 2.27 (d, J = 21.1 Hz, 2H). UPLC-MS (ESI) calculated for C 22 H 18 ClN4O5[M+H] + :453.85, measured value:453.23.

[0288] Example 28: 3-(7-((2-(4-methoxyphenyl)oxazol-5-yl)methoxy)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (D28)

[0289] The synthesis method was similar to that of Example 26 to obtain compound D28. 1 H NMR (500 MHz, DMSO) δ 11.04 (s, 1H), 10.69 (s, 1H), 7.61–7.55 (m, 2H), 7.15–7.05 (m, 3H), 7.05–6.97 (m, 3H), 6.96 (s, 1H), 5.40–5.31 (m, 2H), 5.03 (s, 1H), 3.80 (s, 3H), 2.26 (d, J = 11.0 Hz, 1H), 2.00 (dd, J = 16.0, 8.0 Hz, 2H). UPLC-MS (ESI) requires C 23 H 21 N4O6[M+H] + :449.14, measured value:449.35.

[0290] Example 29: N-(4-chlorobenzyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D29)

[0291] Compound 29-1 (1 g, 4.44 mmol) was dissolved in 20 mL of DMF in a round-bottom flask. Compound 29-2 (1.57 g, 6.67 mmol) and HATU (5 g, 13.32 mmol) were added and stirred for 3 minutes. DIPEA (5.0 g, 22.2 mmol) was then added and stirred at room temperature for 4 hours. After completion of the reaction, the product was dissolved in ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Purification by column chromatography afforded 1.71 g of product 29-3 in a 94% yield.

[0292] Synthesis of compound 29-4:

[0293] Compound 29-3 (1.71 g, 4.1 mmol) was placed in a round-bottom flask and dissolved in 10 ml of tetrahydrofuran. LiOH hydrate (0.984 g, 24.6 mmol) dissolved in 10 ml of water was added and stirred for 1 hour. After the reaction was completed, the solvent was evaporated under reduced pressure. The yield was 100%.

[0294] Synthesis of compound 29-5:

[0295] Compound 29-4 (2.76 g, 6.74 mmol) was placed in a round-bottom flask and dissolved in methanol by stirring. Tetrahydroxydiboron (3.025 g, 33.75 mmol) was added after ice bath at 0°C. Sodium hydroxide (2.7 g, 67.5 mmol) dissolved in methanol was added. After 10 minutes, the mixture was heated at 40°C for 4 hours. After the reaction was completed, the mixture was extracted with ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure and then column chromatography was performed to obtain 2.4 g of the product with a yield of 98%.

[0296] Synthesis of compound 29-8:

[0297] Take compound 29-5 and compound 29-6, and the equivalent and operation steps are the same as compound 3.

[0298] Synthesis of compound 29-9:

[0299] Compound 29-8 (50 mg, 0.10 mmol) was placed in a round-bottom flask, and 5 mg of acetonitrile was stirred and dissolved. Then, p-toluenesulfonic acid hydrate (16 mg, 0.61 mmol) was added and heated under reflux at 90°C for 4 hours. After the reaction, the solvent was evaporated under reduced pressure and purified by high performance liquid chromatography to obtain 10 mg of compound 29 as a white solid, with a yield of 23%. 1 H NMR (500MHz, DMSO-d6) δ9.43(t,J=6.0Hz,1H),8.75(s,1H),7.96(d,J=8.1Hz,1H),7. 91(d,J=8.7Hz,2H),7.49(s,1H),7.43–7.33(m,4H),5.13(dd,J=5.4,3.0Hz,1H),4.5 0 (d, J = 5.9 Hz, 2H), 2.78 (dt, J = 17.6, 3.8 Hz, 1H), 2.62 (ddd, J = 17.7, 12.6, 5.7 Hz, 1H), 2.47 (dd, J = 8.8, 3.9 Hz, 1H), 2.39 (ddq, J = 14.0, 6.0, 2.8 Hz, 1H). UPLC-MS (ESI) theoretical value is C 20 H 18 ClN4O4[M+H] + :413.09, measured value:413.25.

[0300] Example 30: N-((S)-1-(4-chlorophenyl)ethyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D30)

[0301] The synthesis method was similar to that of Example 29 to obtain compound D30. 1H NMR (500MHz, DMSO-d6) δ11.06(s,1H),10.25(s,1H),9.01(d,J=7.8Hz,1H),7.78(d,J=8.1Hz,1H),7.70(s,1H),7.62(dd,J=8.1,1.4Hz,1H),7.41( q,J=8.7Hz,4H),5.39(dd,J=13.1,5.2Hz,1H),5.16(p,J=7.2Hz,1H),2.9 8–2.92(m,1H),2.69–2.64(m,1H),2.46–2.38(m,1H),2.14–2.09(m,1H), 1.48 (d, J = 7.1 Hz, 3H). UPLC-MS (ESI) theoretical value is C 21 H 20 ClN4O4[M+H] + :427.11, measured value:427.26.

[0302] Example 31: N-((S)-1-(4-chlorophenyl)ethyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D31)

[0303] The synthesis method was similar to that of Example 29 to obtain compound D31. 1 H NMR(500MHz,DMSO-d6)δ11.06(s,1H),8.94(d,J=8.0Hz,1H),7.77(d,J=8.1Hz,1H),7.70(s,1H),7.6 4–7.55(m,1H),7.29(d,J=7.9Hz,2H),7.16–7.10(m,2H),5.39(dd,J=13.1,5.2Hz,1H),5.13(h,J=8. 1,7.6Hz,1H),3.70(s,3H),2.95(ddd,J=17.3,13.6,5.4Hz,1H),2.69–2.62(m,1H),2.49–2.37(m,2H),2.28(s,3H),2.12(dtt,J=12.8,5.4,3.3Hz,1H),1.46(dd,J=11.6,7.0Hz,3H). UPLC-MS (ESI) theoretical value was C 22 H 23 N4O4[M+H] + :407.16, measured value:407.33.

[0304] Example 32: 2-(2,6-dioxopiperidin-3-yl)-3-oxo-N-((R)-1-phenylethyl)-2,3-dihydro-1H-indazole-6-carboxamide (D32)

[0305] The synthesis method was similar to that of Example 29 to obtain compound D32. 1 H NMR (500MHz, DMSO-d6) δ11.07(s,1H),8.99(d,J=8.0Hz,1H),7.78(d,J=8.1Hz,1H),7.71(t,J=1.0Hz,1H) ,7.62(dd,J=8.1,1.4Hz,1H),7.44–7.36(m,2H),7.36–7.29(m,2H),7.27–7.19(m,1H),5.39(dd,J=13.1, 5.2Hz,1H),5.18(p,J=7.3Hz,1H),2.95(ddd,J=17.2,13.7,5.4Hz,1H),2.65(dt,J=17.1,3.9Hz,1H),2.43(qd,J=13.3,4.6Hz,1H),2.12(dtd,J=12.9,5.4,2.5Hz,1H),1.50(d,J=7.1Hz,3H). UPLC-MS (ESI) theoretical value was C 21 H 21 N4O4[M+H] + :393.15, measured value:393.32

[0306] Example 33: 2-(2,6-dioxopiperidin-3-yl)-3-oxo-N-((S)-1-phenylethyl)-2,3-dihydro-1H-indazole-6-carboxamide (D33)

[0307] The synthesis method was similar to that of Example 29 to obtain compound D33. 1H NMR (500MHz, DMSO-d6) δ11.06(s,1H),10.24(s,1H),8.99(d,J=8.0Hz,1H),7.77(dd,J=8.1,0.7Hz ,1H),7.71(t,J=1.1Hz,1H),7.62(dd,J=8.2,1.4Hz,1H),7.44–7.38(m,2H),7.37–7.31(m,2H),7.2 7–7.20 (m, 1H), 5.39 (dd, J = 13.2, 5.2 Hz, 1H), 5.18 (p, J = 7.2 Hz, 1H), 2.95 (ddd, J = 17.1, 13.6, 5.4 Hz, 1H), 2.69–2.62 (m, 1H), 2.43 (qd, J = 13.2, 4.5 Hz, 1H), 1.50 (d, J = 7.1 Hz, 3H). UPLC-MS (ESI) theoretical value was C 21 H 21 N4O4[M+H] + :393.15, measured value:393.32

[0308] Example 34: N-((R)-1-(4-chlorophenyl)ethyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D34)

[0309] The synthesis method was similar to that of Example 29 to obtain compound D34. 1 H NMR (500 MHz, DMSO-d6) δ 11.07 (s, 1H), 10.25 (s, 1H), 9.01 (d, J = 7.9 Hz, 1H), 7.89–7.58 (m, 3H), 7.41 (qd, J = 8.5, 1.5 Hz, 4H), 5.39 (dd, J = 13.1, 5.2 Hz, 1H), 5.17 (pd, J = 7.2, 3.0 Hz, 1H), 2.95 (ddd, J = 17.3, 13.7, 5.4 Hz, 1H), 2.71–2.61 (m, 1H), 2.41 (td, J = 13.3, 4.6 Hz, 1H), 1.48 (d, J = 7.1 Hz, 3H). UPLC-MS (ESI) required C 21 H 20 ClN4O4[M+H] + :427.11, measured value:427.37

[0310] Example 35: 2-(2,6-dioxopiperidin-3-yl)-3-oxo-N-((S)-2,2,2-trifluoro-1-phenylethyl)-2,3-dihydro-1H-indazole-6-carboxamide (D35)

[0311] The synthesis method was referred to Example 29 to obtain compound D35. The theoretical value of UPLC-MS (ESI) was C 21 H 20 ClN4O4[M+H] + :447.12, measured value:447.30.

[0312] Example 36: 2-(2,6-dioxopiperidin-3-yl)-N-(4-methylbenzyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D36)

[0313] The synthesis method was similar to that of Example 29 to obtain compound D36. 1 H NMR(500MHz,DMSO-d6)δ9.34(t,J=6.0Hz,1H),8.77–8.72(m,1H),7.95(d,J=8.1Hz,1H),7. 90(dd,J=8.2,1.4Hz,1H),7.87(s,1H),7.53–7.45(m,1H),7.25–7.20(m,2H),7.18–7.10(m ,2H),5.12(dd,J=5.4,3.0Hz,1H),4.47(d,J=6.0Hz,2H),2.89(s,1H),2.78(dt,J=17.5,3.9Hz,1H),2.65–2.59(m,1H),2.40(dt,J=6.1,2.8Hz,1H),2.28(s,3H). UPLC-MS (ESI) theoretical value was C 21 H 21 N4O4[M+H] + :393.15, measured value:393.15.

[0314] Example 37: 2-(2,6-dioxopiperidin-3-yl)-3-oxo-N-((R)-2,2,2-trifluoro-1-phenylethyl)-2,3-dihydro-1H-indazole-6-carboxamide (D37)

[0315] The synthesis method was similar to that of Example 29 to obtain compound D37. 1H NMR (500 MHz, DMSO-d6) δ 11.07 (s, 1H), 10.32 (s, 1H), 9.71 (s, 1H), 7.71 (d, J = 7.2 Hz, 4H), 7.45 (q, J = 7.8, 6.8 Hz, 6H), 6.07 (s, 1H), 5.41 (s, 1H), 3.69 (s, 5H), 2.95 (s, 2H), 2.13 (s, 2H). UPLC-MS (ESI) calculated for C 21 H 18 F3N4O4[M+H] + :447.12, measured value:447.34.

[0316] Example 38: N-((S)-1-(2,4-difluorophenyl)ethyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D38)

[0317] The synthesis method was similar to that of Example 29 to obtain compound D38. 1 H NMR (500MHz, DMSO-d6) δ9.25 (dd, J=12.4, 7.5Hz, 1H), 8.70 (dd, J=3.8, 1.2Hz, 1H), 8.01–7. 85(m,2H),7.62–7.43(m,2H),7.22(dddd,J=15.4,12.3,6.2,2.5Hz,1H),7.09(tt,J=8.7,2. 7Hz,1H),5.43–5.29(m,2H),2.88(dt,J=17.0,5.1Hz,1H),2.76(ddd,J=17.3,10.3,4.5Hz,1H),2.66–2.58(m,1H),2.41–2.36(m,1H),1.50(dd,J=7.1,3.8Hz,3H). UPLC-MS (ESI) theoretical value is C 21 H 19 F2N4O4[M+H] + :429.13, measured value:429.31.

[0318] Example 39: N-((S)-1-(3-chlorophenyl)ethyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D39)

[0319] The synthesis method was similar to that of Example 29 to obtain compound D39. 1H NMR(500MHz,DMSO-d6)δ9.20(d,J=7.8Hz,1H),8.71(s,1H),7.97(d,J=8.1Hz,1H),7.91(dd,J=8.1 ,1.4Hz,1H),7.87(s,1H),7.48(d,J=4.6Hz,2H),7.38(d,J=4.8Hz,2H),7.32(qd,J=4.2,2.0Hz,1H) ,5.19(p,J=7.2Hz,1H),5.13(dd,J=5.5,2.9Hz,1H),3.06(s,1H),2.78(dt,J=17.4,3.8Hz,1H),2.62(dd,J=11.4,6.4Hz,1H),2.40(dt,J=6.9,3.5Hz,1H),1.50(d,J=7.0Hz,3H). UPLC-MS (ESI) theoretical value was C 21 H 20 ClN4O4[M+H] + :427.11, measured value:427.33.

[0320] Example 40: N-(tert-butyl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D40)

[0321] The synthesis method was similar to that of Example 29 to obtain compound D40. 1 H NMR (500 MHz, DMSO-d6) δ 10.33 (s, 1H), 8.63–8.56 (m, 1H), 8.13 (d, J = 13.6 Hz, 1H), 7.95–7.87 (m, 1H), 7.90–7.81 (m, 1H), 5.31 (t, J = 5.4 Hz, 1H), 2.77 (q, J = 5.7 Hz, 1H), 2.69–2.58 (m, 1H), 2.41–2.30 (m, 1H), 2.29–2.16 (m, 1H), 1.43–1.34 (m, 9H). UPLC-MS (ESI) calculated for C 17 H 21 N4O4[M+H] + :345.15, measured value:345.38.

[0322] Example 41: N-((R)-6-chlorochroman-4-yl)-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D41)

[0323] The synthesis method was similar to that of Example 29 to obtain compound D41. 1 H NMR (500MHz, DMSO-d6) δ10.87(s,1H),9.28(dd,J=12.8,8.0Hz,1H),8.77(t,J=1.7Hz,1H ),8.03–7.80(m,2H),7.38–7.11(m,2H),6.94–6.71(m,1H),5.31(q,J=8.4,7.5Hz,1H),4 .30 (qdd, J = 10.7, 6.8, 3.3 Hz, 2H), 2.89 (dt, J = 17.0, 5.1 Hz, 1H), 2.77 (ddd, J = 12.5, 9.1, 4.6 Hz, 1H), 2.70–2.56 (m, 1H), 2.43–2.34 (m, 1H), 2.18–2.06 (m, 2H). UPLC-MS (ESI) theoretical value was C 22 H 20 ClN4O5[M+H] + :455.10, measured value:455.29.

[0324] Example 42: 2-(2,6-dioxopiperidin-3-yl)-N-((R)-8-methylchroman-4-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D42)

[0325] The synthesis method was similar to that of Example 29 to obtain compound D42. 1 H NMR(500MHz,DMSO-d6)δ9.24(dd,J=12.0,8.1Hz,1H),8.76(s,1H),8.01–7.84(m,2H),7.09–6 .98(m,2H),6.79(dd,J=8.0,6.3Hz,1H),5.30(q,J=6.8Hz,1H),5.13(dd,J=5.4,2.9Hz,1H),4. 33 (dddd, J = 33.2, 10.7, 7.3, 3.3 Hz, 2H), 2.92–2.83 (m, 1H), 2.77 (dt, J = 17.4, 4.3 Hz, 1H), 2.61 (dd, J = 11.3, 6.5 Hz, 1H), 2.40 (dt, J = 6.2, 3.0 Hz, 1H), 2.18–2.05 (m, 5H). UPLC-MS (ESI) theoretical value was C 23 H 23 N4O5[M+H] + :435.16, measured value:435.29.

[0326] Example 43: N-butyl-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D43)

[0327] The synthesis method was similar to that of Example 29 to obtain compound D43. 1 H NMR(500MHz,DMSO-d6)δ8.77(t,J=5.7Hz,1H),8.69(t,J=1.0Hz,1H),7.95–7.91(m,1H ),7.90–7.83(m,2H),7.49(s,1H),5.12(dd,J=5.4,3.0Hz,1H),3.31–3.28(m,2H),2.78 (dd, J = 17.4, 4.0 Hz, 1H), 2.65–2.59 (m, 1H), 2.49–2.43 (m, 1H), 2.41–2.36 (m, 1H), 1.54 (tt, J = 7.8, 6.3 Hz, 3H), 1.39–1.28 (m, 3H), 0.92 (t, J = 7.3 Hz, 4H). UPLC-MS (ESI) theoretical value is C 17 H 21 N4O4[M+H] + :345.15, measured value:345.41.

[0328] Example 44: (S)-2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazol-6-yl)methyl)-2,2-difluoroacetamide (D44)

[0329] Step 1:

[0330] Add methyl 4-((bis(tert-butoxycarbonyl)amino)methyl)-2-nitrobenzoate (300 mg, 0.73 mmol) to dichloromethane (2 mL), slowly add 4 mol / L hydrochloric acid in 1,4-dioxane (6 mL), and react at room temperature for 12 hours. After the reaction is complete, concentrate the mixture and proceed directly to the next step.

[0331] Step 2:

[0332] Methyl 4-(aminomethyl)-2-nitrobenzoate hydrochloride (246 mg, 1 mmol), 2-(4-chlorophenyl)-2,2-difluoroacetic acid (227 mg, 1.1 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (418 mg, 1.1 mmol) were added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (1.39 mL, 8 mmol) was added. The mixture was reacted at room temperature for 12 hours. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 100 mg of the product as a colorless oil with a yield of 25%. The theoretical value of UPLC-MS (ESI) was C 17 H 13 ClF2N2O5[M+H] + :399.75, measured value: 399.66.

[0333] Step 3:

[0334] The compound 4-((2-(4-chlorophenyl)-2,2-difluoroacetylamino)methyl)-2-nitrobenzoic acid methyl ester (100 mg, 0.25 mmol) was added to tetrahydrofuran (3 mL), and an aqueous solution (3 mL) of lithium hydroxide monohydrate (16 mg, 0.376 mmol) was added. The reaction was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure and dried to obtain 97 mg of the product, which was used directly in the next reaction without purification.

[0335] Step 4:

[0336] Compound 4-((2-(4-chlorophenyl)-2,2-difluoroacetylamino)methyl)-2-nitrobenzoic acid lithium salt (97 mg), (S)-4,5-diamino-5-oxopentanoic acid tert-butyl ester monohydrochloride (60 mg, 0.25 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (95 mg, 0.25 mmol) were added to N,N-dimethylformamide (3 mL), and N,N-diisopropylethylamine (0.131 mL, 0.75 mmol) was added dropwise. The reaction was allowed to react at room temperature for 12 hours. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 126 mg of the product with a yield of 89%. The theoretical value of UPLC-MS (ESI) was C 25 H 27 ClF2N4O7[M+H] + :568.96, measured value: 568.91.

[0337] Step 5:

[0338] Compound (S)-5-amino-4-(4-((2-(4-chlorophenyl)-2,2-difluoroacetylamino)methyl)-2-nitrobenzamide)-5-oxopentanoic acid tert-butyl ester (30 mg, 0.0.053 mmol) was dissolved in methanol (3 mL). Tetrahydroxydiboron (24 mg, 0.264 mmol) was added at 0°C, followed by a methanol solution (3 mL) of sodium hydroxide (20 mg, 0.5 mmol). The mixture was reacted at 0°C for 10 minutes, then the temperature was raised to 40°C and the reaction was allowed to proceed overnight. After TLC monitoring of the reaction completion, saturated ammonium chloride solution was added to quench the reaction, and ethyl acetate was added for extraction. The organic phase was separated, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 19 mg of the product with a yield of 67%. The theoretical value of UPLC-MS (ESI) was C 25 H 27 ClF2N4O5[M+H] + :537.96, measured value: 537.88.

[0339] Step 6:

[0340] Compound (S)-tert-butyl 5-amino-4-(6-((2-(4-chlorophenyl)-2,2-difluoroacetylamino)methyl)-3-oxo-1,3-dihydro-2H-indazol-2-yl)-5-oxopentanoate (19 mg, 0.035 mmol) and p-toluenesulfonic acid monohydrate (40 mg, 0.212 mmol) were added to acetonitrile (2 mL) and reacted at 80°C overnight. The reaction solution was concentrated and purified by HPLC to obtain 7 mg of the product in a yield of 42%. The theoretical value of UPLC-MS (ESI) was C 21 H 17 ClF2N4O4[M+H] + :463.84, measured value: 463.36. 1 H NMR(500MHz,DMSO)δ10.76(s,1H),9.78(s,1H),9.43(s,1H),8.18(s,1H),7.84(s,1H),7.79(d,J=7.9Hz,1H),7.61(s,2H),7.45(s,1H) ,7.30(d,J=9.1Hz,1H),5.08(d,J=2.9Hz,1H),4.49(d,J=6.1Hz,2H),2.74(d,J=17.7Hz,1H),2.64–2.57(m,2H),2.36(d,J=1.9Hz,1H).

[0341] Example 45: (S)-1-(3-chloro-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazol-6-yl)methyl)urea (D45)

[0342] Step 1:

[0343] Methyl 4-(aminomethyl)-2-nitrobenzoate hydrochloride (180 mg, 0.73 mmol) and 3-chloro-4-methylphenyl isocyanate (135 mg, 0.80 mmol) were added to acetonitrile (8 mL), and triethylamine (0.507 mL, 3.65 mmol) was added. The reaction system was heated to 40°C and reacted for 6 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, diluted with water, and extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 247 mg of the product as a white solid with a yield of 89%. The theoretical value of UPLC-MS (ESI) was C 16 H 14 ClN3O5[M+H] + :364.75, measured value: 364.77.

[0344] Step 2:

[0345] The compound (247 mg, 0.65 mmol) was added to tetrahydrofuran (3 mL), and an aqueous solution (2 mL) of lithium hydroxide monohydrate (41 mg, 0.98 mmol) was added. The mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure and dried to obtain 241 mg of the product, which was used directly in the next reaction without purification.

[0346] Compound 4-((3-(3-chloro-4-methylphenyl)ureido)methyl)-2-nitrobenzoic acid lithium salt (241 mg), (S)-4,5-diamino-5-oxopentanoic acid tert-butyl ester monohydrochloride (171 mg, 0.72 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (274 mg, 0.72 mmol) were added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.34 mL, 1.95 mmol) was added dropwise. The reaction was allowed to react at room temperature for 12 hours. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 322 mg of the product with a yield of 90%. The theoretical value of UPLC-MS (ESI) was C 24 H 28 ClN5O7[M+H] + :534.97, measured value: 534.89.

[0347] Step 3:

[0348] Compound (S)-tert-butyl 5-amino-4-(4-((3-(3-chloro-4-methylphenyl)ureido)methyl)-2-nitrobenzamide)-5-oxopentanoate (358 mg, 0.67 mmol) was added to methanol (5 mL). Tetrahydroxydiboron (300 mg, 3.35 mmol) was added at 0°C, followed by a methanol solution (5 mL) of sodium hydroxide (268 mg, 6.7 mmol). The mixture was allowed to react at 0°C for 10 minutes, then raised to 40°C and allowed to react overnight. After TLC monitoring, the reaction was quenched by adding saturated ammonium chloride solution, extracted with ethyl acetate, and separated. The organic phase was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain 272 mg of the product, 81%. The theoretical value of UPLC-MS (ESI) was C 25 H 30 ClN5O5[M+H] + :517.00, measured value: 517.03.

[0349] Step 4:

[0350] Compound (S)-tert-butyl 5-amino-4-(6-((3-(3-chloro-4-methylphenyl)ureido)methyl)-3-oxo-1,3-dihydro-2H-indazol-2-yl)-5-oxopentanoate (272 mg, 0.53 mmol) and p-toluenesulfonic acid monohydrate (601 mg, 3.16 mmol) were added to acetonitrile (mL) and reacted at 80°C overnight. The reaction solution was concentrated and purified by HPLC to obtain 60 mg of the product in a yield of 26%. The theoretical value of UPLC-MS (ESI) was C 21 H 20 ClN5O4[M+H] + :442.87, measured value: 442.91.

[0351] Example 46: N-Benzyl-2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D46)

[0352] The synthesis method was similar to that of Example 29 to obtain compound D46. 1H NMR (500MHz, DMSO) δ11.06(s,1H),10.26(s,1H),9.21(t,J=5.9Hz,1H),7.77(d,J=8.1Hz, 1H),7.72(s,1H),7.62(d,J=8.2Hz,1H),7.33(d,J=4.7Hz,4H),7.25(dd,J=8.8,4.4Hz,1H ),5.39(dd,J=13.1,5.1Hz,1H),4.50(d,J=5.9Hz,2H),2.93(dd,J=22.3,8.7Hz,1H),2.65(d,J=12.5Hz,1H),2.40(dd,J=19.8,15.6Hz,1H),2.14–2.06(m,1H). UPLC-MS (ESI) theoretical value is C 20 H 19 N4O4[M+H] + :379.13, measured value:379.25.

[0353] Example 47: 2-(2,6-dioxopiperidin-3-yl)-N-((S)-1-(4-ethylphenyl)ethyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxamide (D47)

[0354] The synthesis method was similar to that of Example 29 to obtain compound D47. 1 H NMR (500MHz, DMSO) δ11.06(s,1H),10.22(s,1H),8.93(d,J=8.0Hz,1H),7.76(d,J=8.2Hz,1H),7.69(s ,1H),7.61(d,J=8.2Hz,1H),7.30(d,J=8.1Hz,2H),7.16(d,J=8.1Hz,2H),5.38(dd,J=13.1,5.2Hz,1H) ,5.14(p,J=7.1Hz,1H),2.99–2.89(m,1H),2.64(d,J=16.6Hz,1H),2.57(q,J=7.6Hz,2H),2.41(qd,J=13.3,4.5Hz,1H),2.15–2.05(m,1H),1.47(d,J=7.1Hz,3H),1.16(t,J=7.6Hz,3H). UPLC-MS (ESI) theoretical value was C 23 H 25 N4O4[M+H] + :421.18, measured value:421.16.

[0355] 2. Experimental Examples

[0356] The present invention also tested the activity of the polysubstituted isoindoline compounds described in the examples against three major hematological tumor cell lines: the multiple myeloma cell line (MM.1S), the mantle cell lymphoma cell line (Mino), and the acute myeloid leukemia cell line (MV-4-11). The cell proliferation inhibition activity of these three representative cell lines was tested. All experimental materials required for the pharmacological experiments were commercially available, unless otherwise specified.

[0357] 1. Effects of compounds on MM.1S cell proliferation

[0358] MM.1S cells were cultured with 1640 plus 10% fetal bovine serum and collected. The cell concentration was diluted according to the 7-day action time, and 180ul of cell suspension was added to each well of a 96-well cell plate to make the cell number 20,000. 20ul of DMSO with a final concentration of 0.2% was added to the control cell wells. The compound was diluted 5 times from the 10mM stock solution, and 20ul was also added to the compound cell wells (DMSO final concentration was 0.2%). The cells were placed in a 37°C, 5% CO2 incubator and incubated for 7 days. After preparing the reaction solution according to the MTS kit (Promega, G5430), 20μL was added to each well and incubated in a 37°C, 5% CO2 incubator for 3-4 hours. The 490nm absorbance value was read using an enzyme-labeled plate, and the 690nm absorbance value was used as the background value, with OD490-OD690 as the final raw data. The inhibition rate of the compound was calculated as follows: Inhibition rate = (OD DMSO -OD 化合物 ) / (OD DMSO -O 空白 ) × 100%. The IC50 of the proliferation inhibition of the compound was fitted by Graph Pad Prism 5.0. The experiment was repeated three times, and the mean and standard deviation were calculated using three parallel experiments each time.

[0359] The results of cell activity test are shown in Table 1 below: A represents cell activity IC 50 <200nM, B indicates cell viability 200nM <IC 50 <1μM, C indicates cell viability 1μM <IC 50 <10μM, D represents the IC value of cell viability 50 >10μM.

[0360] Table 1. Inhibitory activity of compounds on MM.1S cell proliferation

[0361] Based on the above test results of the cell growth inhibitory activity of the compounds, some of the embodiment compounds of the present invention have good inhibitory activity on the growth of multiple myeloma MM.1S cells, and the activity of some compounds is better than that of the listed drugs lenalidomide or pomalidomide. On the other hand, the development of these structurally diverse compounds provides more optional material bases and new structures for obtaining drug molecules with higher activity and molecules with better pharmaceutical properties. Therefore, the compounds of the present invention can be used to prevent, treat and regulate CRBN (CRL4 CRBN Diseases associated with E3 ubiquitin ligase) activity, such as multiple myeloma or, without limitation, other potential tumor diseases, pain, neurological diseases, and immune system diseases.

[0362] 2. Inhibitory activity of compounds on Mino cell proliferation

[0363] Mino cells were cultured in 1640 plus 15% fetal bovine serum and harvested. Cells were diluted to a concentration of 8,000 cells per well of a 96-well plate. 10 μl of the cell suspension was added to the control wells to a final concentration of 0.2% DMSO. Compounds were serially diluted 5-fold from a 10 mM stock solution and 10 μl was added to the compound wells (final DMSO concentration of 0.2%). Cells were incubated at 37°C, 5% CO2 for 3 days. The reaction mixture was prepared according to the MTS kit (Promega, G5430), and 20 μl was added to each well. The cells were incubated at 37°C, 5% CO2 for 3-4 hours.

[0364] The absorbance value at 490 nm was read using an enzyme labeling plate, and the absorbance value at 690 nm was used as the background value. The final raw data was OD490-OD690. The inhibitory rate of the compound was calculated as follows: Inhibition rate = (OD DMSO -OD 化合物 ) / (OD DMSO -O 空 白 )×100%. IC of the compound for proliferation inhibition 50 The data were fitted by Graph Pad Prism 5.0. The experiment was repeated three times, and the mean and standard deviation were calculated using three parallel experiments each time.

[0365] The results of cell activity test are shown in Table 2 below: A represents cell activity IC 50 <200nM, B indicates cell viability 200nM <IC 50 <1μM, C indicates cell viability 1μM <IC 50 <10μM, D represents the IC value of cell viability 50 >10μM.

[0366] Table 2. Inhibitory activity of compounds on Mino cell proliferation

[0367] Based on the test results of the cell growth inhibitory activity of some of the above-mentioned example compounds, it was found that some example compounds of the present invention have good inhibitory activity on the growth of mantle cell lymphoma Mino cells, and the activity of most of the compounds is better than that of the marketed drugs lenalidomide or pomalidomide. On the other hand, the development of these structurally diverse compounds provides an optional material basis and molecular source for obtaining more active drug molecules and molecules with better pharmaceutical properties. Therefore, the compounds of the present invention broaden the scope of application of amide drugs in the treatment of hematological tumor diseases, and can be used to expand to other indications of hematological tumors, such as as active molecules for mantle cell lymphoma diseases, and as a use for preparing drugs or diagnostic reagents for preventing or treating such diseases. Therefore, the compounds of the present invention can be used as potent new CRBN modulators for the prevention, treatment and regulation of CRBN (CRL4 CRBN Diseases associated with decreased E3 ubiquitin ligase (E3 ubiquitin ligase) activity, such as multiple myeloma, mantle cell lymphoma, or non-limitingly other potential tumor diseases, pain, neurological diseases, and immune system diseases.

[0368] 3. Inhibitory activity of compounds on MV-411 cell proliferation

[0369] MV-4-11 cells were cultured with IMDM plus 10% fetal bovine serum and collected. The cell concentration was diluted according to the 7-day action time, and 180ul of cell suspension was added to each well of a 96-well cell plate to make the cell number 2000. 20ul of DMSO with a final concentration of 0.2% was added to the control cell wells. The compound was diluted 5-fold from the 10mM stock solution, and 20ul was also added to the compound cell wells (DMSO final concentration was 0.2%). The cells were placed in a 37°C, 5% CO2 incubator and incubated for 7 days. After preparing the reaction solution according to the MTS kit (Promega, G5430), 20μL was added to each well and incubated in a 37°C, 5% CO2 incubator for 3-4 hours. The absorbance value at 490nm was read using an enzyme-labeled plate, and the absorbance value at 690nm was used as the background value. The final raw data was OD490-OD690. The inhibition rate of the compound was calculated as follows: Inhibition rate = (OD DMSO -OD 化合物 ) / (OD DMSO -O 空白 ) × 100%. The IC50 of the proliferation inhibition of the compound was fitted by Graph Pad Prism 5.0. The experiment was repeated three times, and the mean and standard deviation were calculated using three parallel experiments each time.

[0370] The results of the cell activity test are shown in Table 3 below: A represents cell activity IC 50 <200nM, B indicates cell viability 200nM <IC 50 <1μM, C indicates cell viability 1μM≤IC 50 ≤20μM, D represents cell activity IC 50 >20μM.

[0371] Table 3. Inhibitory activity of compounds on MV-4-11 cell proliferation

[0372] Based on the results of the cell growth inhibition activity test of some of the above-mentioned compounds in the acute myeloid leukemia cell line (MV-4-11), it was found that some of the compounds of the present invention have very good inhibitory activity against acute leukemia cells MV-4-11. 50 At the nanomolar level, the best IC activity of the compounds tested in the table 50 can reach <100nM; however, the positive compounds, whether they are the marketed drugs lenalidomide or pomalidomide, or the compound CC-220 currently under clinical investigation, have low inhibitory activity (IC 50 ) are all greater than 20 μM. Therefore, the compounds of the present invention, in which the amine nucleus was modified into an indazolone nucleus, achieved unexpected results in the acute myeloid leukemia cell line (MV-4-11), far outperforming marketed drugs and those under clinical development in terms of cell activity.

[0373] 4.CRBN / DDB1 binding TR-FRET assay

[0374] 1. Experimental steps

[0375] 1. Take 0.1 μL of reference solution and place it in a 384-well reaction plate. The final detection concentration is 100 μM. Dilute 3 times and plate 10 points.

[0376] 2. Add 2.5 μL of CRBN / DDB1 to the 384-well reaction plate and centrifuge at 1000 rpm for 1 minute.

[0377] 3. Add 2.5 μL of Thalidomide-BODIPY to the 384-well reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 15 minutes.

[0378] 4. Add 5 μL of His-Tb to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 60 minutes.

[0379] 5. Read the TR-FRET signal using BMG (ratio 520 / 490 nm).

[0380] The inhibition rates of the compounds on CRBN / DDB1 binding are shown in Table 4 below: A indicates an inhibition rate > 90%, B indicates an inhibition rate of 50% < 90%, and C indicates an inhibition rate < 50%.

[0381] Table 4. Inhibition rate of compounds on CRBN / DDB1 binding

[0382] Based on the test results of the inhibition rate of CRBN / DDB1 binding of some of the above example compounds at a concentration of 10 micromolar, it was found that most of the example compounds of the present invention have good inhibitory activity on CRBN / DDB1 binding, and most of the compounds have inhibitory activity better than or equivalent to that of lenalidomide.

[0383] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof: in, R1 is hydrogen, methyl or ethyl; R2 is hydrogen or fluorine; R3 is hydrogen, deuterium or fluorine; L is none, -CH2-, S, -CO-, -CH(CH3)-; X-A1 combination is -(CH2) n -O-CH2-A1, -(CH2) n -NHCONH-A1, -(CH2) n -NHCOCF2-A1, -CONH-(CH2) n -A1, -CONH-CO-A1, -CONHCH(CH3)-A1, -CONHCH(CF3)-A1; n is 0 or 1; When X-A1 is combined with -(CH2) n -O-CH2-A1 or, L is none, -CH2-, S, -CO-, -CH(CH3)-; A1 is selected from: halogen-substituted and unsubstituted phenyl, halogen-substituted and unsubstituted pyridyl, and five-membered heteroaryl containing N, O, and S; A2 is selected from: phenyl, pyridyl, piperazinyl, piperidinyl, morpholinyl, azetidinyl; wherein the phenyl, pyridyl, piperazinyl, piperidinyl, morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, nitro, hydroxyl, carboxyl, aminocarbonyl, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen substituted C1-C6 alkyl, halogen substituted C1-C6 alkoxy, substituted or unsubstituted phenyl, substituted or unsubstituted Pyridyl, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy substituted by C1-C6 alkoxy; wherein the substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy; When X-A1 is combined with -(CH2) n -NHCONH-A1 or -(CH2) n -NHCOCF2-A1, L and A2 are both absent; A1 is selected from: phenyl, wherein the phenyl is substituted by one or more groups selected from the following: hydrogen, halogen, cyano, C1-C6 alkyl; When the X-A1 combination is -CONHCH(CH3)-A1, -CONH-(CH2) n When -A1, -CONH-CO-A1 or -CONHCH(CF3)-A1, L and A2 are not present at the same time; A1 is selected from: phenyl, pyridyl, 5-10 membered heteroaryl, C3-C10 cycloalkyl, 4-6 membered heterocyclyl, 4-6 membered heterocyclylphenyl, C1-C6 alkyl; the aforementioned C6-10 aryl, 5-10 membered heteroaryl, C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl is substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy.

2. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: The compounds have structures shown in formula (I-1)-(I-11): Wherein, R1 is hydrogen, methyl or ethyl; R2 is hydrogen or fluorine; R3 is hydrogen, deuterium or fluorine; R4 is H or halogen; n = 0 or 1; R5 is selected from phenyl, pyridyl or morpholinyl, wherein the phenyl, pyridyl or morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen substituted C1-C6 alkyl, halogen substituted C1-C6 alkoxy; A2 is selected from: phenyl, pyridyl; wherein the phenyl, pyridyl is optionally substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy substituted by C1-C6 alkoxy.

3. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: The compounds have structures shown in formula (I-29)-(I-30): Wherein, R1 is hydrogen, methyl or ethyl; R2 is hydrogen or fluorine; R3 is hydrogen, deuterium or fluorine; R 11 is hydrogen or methyl; Each q is independently 0 or 1; Z is N or -CH-; R5 is selected from phenyl, pyridyl or morpholinyl, wherein the phenyl, pyridyl or morpholinyl is optionally substituted by one or more groups selected from the following: hydrogen, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen substituted C1-C6 alkyl, halogen substituted C1-C6 alkoxy; A2 is selected from: phenyl, pyridyl; wherein the phenyl, pyridyl is optionally selected from the following one or more groups Substitution: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy, hydroxy-substituted C1-C6 alkoxy, C1-C6 alkoxy substituted by C1-C6 alkoxy.

4. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: The compounds have structures shown in formula (I-12)-(I-13): Wherein, R1 is hydrogen, methyl or ethyl; R2 is hydrogen or fluorine; R3 is hydrogen, deuterium or fluorine; n is 0 or 1; R6 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl; m is 0, 1, 2, 3, 4 or 5.

5. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: The compounds have structures shown in formula (I-31)-(I-32): Wherein, R1 is hydrogen, methyl or ethyl; R2 is hydrogen or fluorine; R3 is hydrogen, deuterium or fluorine; R6 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl; m is 0, 1, 2, 3, 4 or 5.

6. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: The compounds have structures shown in formula (I-20)-(I-26): Wherein, R1 is hydrogen, methyl or ethyl; R2 is hydrogen or fluorine; R3 is hydrogen, deuterium or fluorine; R9 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy; p is 0, 1, 2, 3, 4 or 5; A1 is selected from: phenyl, pyridyl, substituted or unsubstituted C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl; the aforementioned phenyl, pyridyl, substituted or unsubstituted C3-C10 cycloalkyl, 4-6 membered heterocyclyl, C1-C6 alkyl is substituted by one or more groups selected from the following: hydrogen, deuterium, halogen, cyano, 4-10 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, C3-C8 heterocyclyloxy, C3-C8 cycloalkyloxy.

7. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: Selected from: Selected from:

8. The compound according to claim 1, its pharmaceutically acceptable salt, tautomer, stereoisomer, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof, characterized in that: The compounds are the following compounds:

9. A method for preparing the compound of claim 1, wherein the method is selected from one of the following methods: Synthesis method 1: Step 1-1: Compound 1A and compound 1B undergo nucleophilic substitution reaction to obtain compound 1C; Step 1-2: Compound 1C and compound 1D undergo nucleophilic substitution reaction to obtain compound 1E; Step 1-3: Compound 1E is hydrolyzed with alkaline to obtain compound 1F; Step 1-4: Compound 1F and 1G are subjected to condensation reaction to obtain compound 1H; Step 1-5: Compound 1H reacts with tetrahydroxydiboron and sodium hydroxide to obtain compound 1I Step 1-6: Compound 1I is subjected to p-toluenesulfonic acid reflux reaction to obtain compound 1J; R5 is the same as defined in claim 2; Synthesis method 2: Step 2-1: Compound 2A reacts with compound 2B in the presence of N,N'-carbonyldiimidazole and triethylamine to obtain compound 2C; Step 2-2: Compound 2A and compound 2D are reacted in the presence of HATU and N,N-diisopropylethylamine to obtain compound 2E; R2, R3, R6, and m are the same as those defined in claim 4; Synthesis method three: Step 3-1: Compound 3A and R8-NH2 undergo condensation reaction to obtain compound 3B; Step 3-2: Compound 3B is hydrolyzed in the presence of lithium hydroxide to obtain compound 3C; Step 3-3: Compound 3C and 1G are subjected to condensation reaction to obtain compound 3D; Step 3-4: Compound 3D reacts with tetrahydroxydiboron and sodium hydroxide to obtain compound 3E; Step 3-5: Compound 3E is subjected to p-toluenesulfonic acid reflux reaction to obtain compound 3F; R8-NH2 is selected from: R9 has the same meaning as defined in claim 6.

10. A pharmaceutical composition comprising a therapeutically effective dose of the compound of claim 1, its tautomers, enantiomers, diastereomers, racemates, isotopic compounds, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof and other pharmaceutically acceptable carriers.

11. The pharmaceutical composition according to claim 10, further comprising one or more selected from the group consisting of dexamethasone, rituximab, trastuzumab, PD-1 inhibitor, PDL-1 inhibitor, pemetrexed, topotecan, doxorubicin, bortezomib, gemcitabine, dacarbazine, clarithromycin, vincristine, cytarabine, prednisone, docetaxel, clofarabine injection, HDAC inhibitor, androgen receptor inhibitor, androgen biosynthesis inhibitor, BTK inhibitor, erythrocyte growth hormone, minocycline, Elotuzumab, Palbociclib, Nivolumab, Pembrolizumab, Panobinostat, Ublituximab, Romidepsin, Eltrombopag, CAR-T and melphalan.

12. Use of the compound according to claim 1, its tautomer, enantiomer, diastereomer, racemate, isotope compound, pharmaceutically acceptable salt, ester, prodrug or hydrate thereof in the preparation of a medicament for preventing or treating a disease associated with CRL4 CRBN E3 ubiquitin ligase.

13. The use according to claim 12, wherein The diseases associated with CRL4 CRBN E3 ubiquitin ligase are cancer, pain, central nervous system diseases and immune system diseases.

14. The use according to claim 11, wherein The disease is a hematological tumor or a solid tumor.

15. The use according to claim 14, wherein The disease is selected from the group consisting of myelodysplastic syndrome, multiple myeloma, mantle cell lymphoma, non-Hodgkin's lymphoma, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, myelofibrosis, Burkitt's lymphoma, Hodgkin's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, ciliary body and chronic melanoma, iris melanoma, recurrent interocular melanoma, T-cell lymphoma, erythroid lymphoma, monocytic and monocytic leukemia, acute myeloid leukemia, central nervous system disease, Systemic lymphoma, meningioma, spinal cord tumor, non-small cell lung cancer, ovarian cancer, skin cancer, renal cell carcinoma, astrocytoma, amyloidosis, complex regional pain syndrome type I, malignant melanoma, radiculopathy, glioblastoma, gliosarcoma, malignant glioma, refractory plasmacytoma, extraocular extension of melanoma, papillary and follicular thyroid cancer, breast cancer, prostate cancer, hepatocellular carcinoma or primary macroglobulinemia, large cell lymphoma, myeloid leukemia, brain tumor, thyroid cancer.

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