Heterocyclic substituted isoindoline compound as well as preparation method, pharmaceutical composition and application thereof

Abstract

The invention relates to a heterocyclic substituted isoindoline compound as well as a preparation method, a pharmaceutical composition and application thereof. Specifically, the invention provides a compound as shown in a formula (I), and pharmaceutically acceptable salts, tautomers or stereoisomers thereof, the invention further discloses application of the compound serving as a molecular glue degradation agent of the CRL4CRBNE3 ubiquitin ligase to drugs for preventing or treating diseases related to the CRL4CRBNE3 ubiquitin ligase.

Description

TECHNICAL FIELD

[0001] The present application relates to a class of heterocyclic substituted isoindoline compounds, their preparation methods, pharmaceutical compositions and uses. The compounds of the present application, as a class of novel CRL4CRBN E3 ubiquitin ligase modulators, can be used as degradation agents for related pathogenic proteins, for the preparation of drugs for the prevention and treatment of diseases related to CRBN. BACKGROUND

[0002] Targeted protein degradation is a rapidly developing and breakthrough drug treatment method. This strategy recruits pathogenic proteins through small molecules, and then rapidly degrades them using the ubiquitin-proteasome pathway, thereby achieving the effect of treating diseases. At present, the targeted protein degradation strategy shows great potential in the treatment of diseases such as cancer, infectious diseases, inflammation and neurodegenerative diseases, especially for the degradation of those with “undruggable” targets. Molecular glue (MG), a class of chemical small molecules that act on the protein-protein interaction interface, can mediate new protein-protein interactions, and together with protein degradation targeting chimera (PROTACs) are collectively referred to as targeted protein degradation technology. The advantage of molecular glue is that it can promote the protein-protein interaction between ubiquitin ligase and target protein, and in theory can degrade proteins without ligand binding, showing better therapeutic effect than traditional small molecule inhibitors. Compared with PROTACs, molecular glue compounds have smaller molecular weight, and are expected to have better pharmacological properties, better membrane permeability, better cell uptake and better blood-brain barrier penetration ability, etc.

[0003] Recent studies have shown that immunomodulatory imide drugs (IMiD) such as lenalidomide exert therapeutic effects through the mechanism of molecular glue. This class of immunomodulatory drugs targets CRL4 CRBN E3 ubiquitin ligase, binds to E3 ubiquitin ligase cereblon (CRBN) to cause a change in the conformation of the protein binding interface, thereby recruiting Ikaros (IKZF1), Aiolos (IKZF3), CK1α, GSPT1 and other proteins for ubiquitination and subsequent degradation by 26S proteasome (Science 2014, 343(6168), 301-305). The downstream effects of these protein degradations mediate the anti-tumor proliferation and immunomodulatory activities of immunomodulatory drugs. Different lenalidomide compounds and CRL4 CRBNE3 ubiquitin ligase complex binding, showing different substrate protein degradation specificity, and thus can be used for the treatment of different indications. For example, lenalidomide, which degrades IKZF1 and IKZF3, is used to treat multiple myeloma (MM); and lenalidomide can also degrade CK1α to treat 5q deletion myelodysplastic syndrome (5qMDS); DKY-709 developed by Novartis causes Helios (IKZF2) degradation for the treatment of solid tumors, and is currently in clinical research (Cell Chem Biol 2023, 30(3), 235-247).

[0004] Although the existing CRBN-binding compounds are similar in chemical structure, they each have different modes of action, ways of degrading substrate proteins, clinical efficacy, and side effects. Therefore, the development of new CRL4 CRBN E3 ubiquitin ligase molecular glue degraders can recruit and degrade new substrate proteins, which can further explore new indications for treatment, improve the selectivity of degradation and reduce side effects, and improve treatment efficacy to meet different clinical needs, and have important research value and new drug development significance. SUMMARY

[0005] The purpose of the present application is to provide a CRL4 CRBN E3 ubiquitin ligase molecular glue degraders and their preparation methods and uses, the compounds of the present application can improve the treatment effect of tumors and reduce side effects, thereby broadening the new indications of CRL4CRBNE3 ubiquitin ligase-based molecular glue degraders.

[0006] In a first aspect, the present application provides a compound of formula (I), a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0007]

[0008] wherein X is CHR3, CR3R4, CO;

[0009] each R3 and R4 is independently hydrogen or C1-C6 alkyl;

[0010] R1 is hydrogen or halogen;

[0011] R2 is hydrogen, fluorine or C1-C6 alkyl;

[0012] Z is (CH2)n, -O-(CH2)n, -NH-(CH2)n;

[0013] n is independently 4, 5 or 6;

[0014] Y is hydrogen, halogen, Ci-C6-alkyl or Ci-C6-haloalkyl;

[0015] Y1is O;

[0016] X1is C-R5or N;

[0017] X2is C-R6or N;

[0018] X3is C-R7or N;

[0019] X4is C-R8or N;

[0020] wherein only one of X1, X2, X3, X4is N;

[0021] R5, R6, R7, R8are each independently selected from the group consisting of hydrogen, halogen, cyano, nitro, hydroxy, carboxyl, aminocarbonyl, Ci-C6-alkyl, halogen-substituted Ci-C6-alkyl, hydroxy-substituted Ci-C6-alkyl, Ci-C6-alkoxy, halogen-substituted Ci-C6-alkoxy.

[0022] In another preferred embodiment, the compound has the structure of formula (I-1):

[0023]

[0024] wherein X is CHR3, CR3R4, CO;

[0025] each R3and R4is independently hydrogen;

[0026] R1is hydrogen or fluorine;

[0027] R2is hydrogen, fluorine or methyl;

[0028] Z is (CH2)n, -O-(CH2)n, -NH-(CH2)n;

[0029] n is 4, 5 or 6;

[0030] Y is hydrogen;

[0031] X1is C-R5or N;

[0032] X2is C-R6or N;

[0033] X3is C-R7or N;

[0034] X4is C-R8or N;

[0035] wherein only one of X1, X2, X3, X4is N;

[0036] R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, hydroxyl, aminocarbonyl, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

[0037] In another preferred embodiment, the compound has the structure shown in formula (I-2):

[0038]

[0039] Where R1 is hydrogen or fluorine;

[0040] Z is (CH2)n or -O-(CH2)n;

[0041] n is 4, 5, or 6;

[0042] Y represents hydrogen;

[0043] X1 is either C-R5 or N;

[0044] X2 is either C-R6 or N;

[0045] X3 is either C-R7 or N;

[0046] X4 is either C-R8 or N;

[0047] Among X1, X2, X3, and X4, only one is N;

[0048] R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

[0049] In another preferred embodiment, the compound has the structure shown in formula (I-3):

[0050]

[0051] R1 is hydrogen or fluorine;

[0052] n is 4, 5, or 6;

[0053] Y represents hydrogen;

[0054] X1 is either C-R5 or N;

[0055] X2 is either C-R6 or N;

[0056] X3 is either C-R7 or N;

[0057] X4 is either C-R8 or N;

[0058] Among X1, X2, X3, and X4, only one is N;

[0059] R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

[0060] In another preferred embodiment, Z is (CH2)n, and n is 4, 5, or 6.

[0061] In another preferred embodiment, R1 is hydrogen or fluorine.

[0062] In another preferred embodiment, each Y is H.

[0063] In another preferred example, X is CH2.

[0064] In another preferred embodiment, Y1 is 0.

[0065] In another preferred embodiment, X3 is C-R7 and any one of X1, X2 and X4 is N.

[0066] In another preferred embodiment, Selected from:

[0067]

[0068] In another preferred embodiment, the compound is selected from the group consisting of:

[0069]

[0070]

[0071]

[0072]

[0073] In a second aspect, the present invention provides a method for preparing the compound described in the first aspect, wherein the method is selected from one of the following methods:

[0074]

[0075] X, X1, X2, X3, X4, Y, R1, and n are the same as those defined in claim 3, wherein X5 is CH2 or O;

[0076] Step 1-1: Compounds 1A and 1B react with potassium carbonate in DMF solution by heating to obtain compound 1C;

[0077] Steps 1-2: Compound 1C, tri-n-butyltin hydrogen and azobisisobutyronitrile were reacted in toluene at 135±10℃ to give compound 1D;

[0078] Steps 1-3: Compound 1D reacts with dioxane hydrochloride at room temperature to give compound 1F;

[0079] Steps 1-4: Compounds 1G and 1F reacted with triethylamine at room temperature to give compound 1H;

[0080] Synthesis Method Two:

[0081]

[0082] X1, X2, X3, X4, n, and Y are the same as those defined in claim 4;

[0083] Step 2-1: Compound 2A and compound 2B react in the presence of N,N-diisopropylethylamine to give compound 2C;

[0084] Step 2-2: Compound 2C reacts with carbon tetrabromide and triphenylphosphine to give compound 2D;

[0085] Steps 2-3: Compounds 2D and 1F reacted with triethylamine at room temperature to give compound 2E;

[0086] In a third aspect, the present invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition comprises the compound of any one of claims 1-6, a pharmaceutically acceptable salt, tautomer or stereoisomer, and a pharmaceutically acceptable carrier.

[0087] In another preferred embodiment, the pharmaceutical composition further comprises one or more other pharmaceutically therapeutically active ingredients that produce a synergistic effect in the prevention or treatment of a specific disease or functional disorder; or reduce or eliminate the toxic side effects produced by one or more other pharmaceutically therapeutically active ingredients in the prevention or treatment of a specific disease or functional disorder.

[0088] In another preferred embodiment, the pharmaceutical composition further comprises one or more selected from dexamethasone, rituximab, trastuzumab, PD-1 inhibitors, PD-L1 inhibitors, pemetrexed, topotecan, doxorubicin, gemcitabine, dacarbazine, clarithromycin, vincristine, cytarabine, azacitidine, prednisone, docetaxel, clofarabine injection, HDAC inhibitors, androgen receptor inhibitors, androgen biosynthesis inhibitors, BTK inhibitors, BCL2 inhibitors, erythrocyte growth hormone, minocycline, elotuzumab, palbociclib, nivolumab, pembrolizumab, panobinostat, ubliximab, romidepsin, eltrombopag, CAR-T, and melphalan.

[0089] In a fourth aspect, the present invention provides a compound as described in the first aspect, a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, or a pharmaceutical composition as described in the second aspect of the present invention, for the preparation of a compound for the prevention or treatment of CRL4. CRBN Use in medicines for diseases related to E3 ubiquitin ligases, preferably, those related to CRL4 CRBN Diseases associated with E3 ubiquitin ligase include cancer, pain, inflammation, central nervous system disorders, or immune system disorders.

[0090] In another preferred embodiment, the cancer is a hematologic malignancy or a solid tumor. Preferably, the hematologic malignancy is selected from: myelodysplastic syndrome, multiple myeloma, mantle cell lymphoma, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, myelofibrosis, Burkitt lymphoma, Hodgkin 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 monocyte leukemia, myeloid leukemia, and central nervous system lymphoma; the solid tumor is selected from: From: Meningioma, spinal cord tumor, lung cancer, ovarian cancer, skin cancer, renal cell carcinoma, astrocytoma, amyloidosis, type I complex regional pain syndrome, malignant melanoma, radiculopathy, glioblastoma, sarcoma, malignant glioma, refractory plasmacytoma, extraocular melanoma, thyroid cancer, breast cancer, prostate cancer, hepatocellular carcinoma or primary macroglobulinemia, adrenocortical carcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colorectal cancer, esophageal cancer, gallbladder cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, uterine cancer, rhabdomyosarcoma, neuroblastoma, pancreatic cancer, testicular cancer.

[0091] In another preferred embodiment, the pain is nociceptive pain, neuropathic pain, visceral pain, migraine, headache, or postoperative pain.

[0092] In another preferred embodiment, the neuropathic pain is a complex regional pain syndrome, malnutrition, or diabetic neuropathy.

[0093] In another preferred embodiment, the central nervous system disease is amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis, or Tourette syndrome.

[0094] In a fifth aspect, the present invention provides a method for preventing or treating CRL4. CRBN A method for treating E3 ubiquitin ligase-related diseases, comprising the steps of: administering to a subject in need a compound as described in the first aspect of the invention, a pharmaceutically acceptable salt thereof, a tautomer, a stereoisomer thereof, or a pharmaceutical composition as described in the third aspect of the invention, thereby preventing or treating said disease.

[0095] In another preferred embodiment, the subject is a human or a non-human mammal, such as a rat or mouse.

[0096] In another preferred embodiment, the disease is a solid tumor, hematologic malignancy, etc.

[0097] It should be understood that, within the scope of this invention, the above-described technical features of this 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. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0098] Through extensive and in-depth research, and through numerous screenings and tests, the inventors have provided a novel CRL4 with excellent activity. CRBN This invention relates to E3 ubiquitin ligase molecular gel-like degrading agents, their preparation methods, pharmaceutical compositions, and uses.

[0099] the term

[0100] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0101] As used herein, when referring to a specific enumerated value, the term “about” means that the value can vary by no more than 1% from the enumerated values. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0102] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0103] As used herein, the term “room temperature” or “normal temperature” refers to a temperature of 4–40°C, preferably 25 ± 5°C.

[0104] When a substituent is described using a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH2O- includes -OCH2-.

[0105] Those skilled in the art will understand that the combinations of substituents contemplated in this invention are those that are stable or chemically feasible. Substitution in the relevant structure in this invention includes both substitution and non-substitution; for example, "optionally" or "optionally" substituted by a substituent means both substitution and non-substitution.

[0106] The statement in this invention that when the number of substituents is greater than 1, the substituents R can be the same or different substituents means that when there are multiple substituents in a certain structure, the combination of substituents R can be selected from a variety of different types of substituents.

[0107] The term "substitution" applies only to sites that can be replaced by a substituent and does not include substitutions that are not achievable with existing chemical knowledge. The term "substitution" refers to the replacement of one or more hydrogen atoms on a specific group by a specific substituent. A specific substituent is a substituent described above or that appears in the examples. Unless otherwise specified, an arbitrarily substituted group may have a substituent selected from a specific group at any substituted site of that group, and the substituents may be the same or different at each position. Cyclic substituents, such as heterocyclic alkyl groups, may be attached to another ring, such as a cycloalkyl group, thereby forming a spirobicyclic system, for example, where the two rings share a single carbon atom.

[0108] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0109] The term "C1-C6 alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms on the chain, including, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, etc.

[0110] The term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1 to 6 carbon atoms, including, without limitation, methoxy, ethoxy, propoxy, isopropoxy, and butoxy.

[0111] The term "aryl" refers to a 6-14 membered all-carbon monocyclic or fused polycyclic group with 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, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring.

[0112] The term "halogen-substituted C1-C6 alkoxy" refers to a straight-chain or branched C1-C6 alkoxy group substituted with one or more halogens, including, without limitation, -OCH2F, -OCHF2, and -OCF3.

[0113] The term "halogen-substituted C1-C6 alkyl" refers to straight-chain or branched C1-C6 alkyl groups substituted with one or more halogens, including, without limitation, 2-bromoethyl, 2-bromopropyl, etc.

[0114] The term "heteroaryl" refers to a 5-14 membered aryl group having 1 to 4 heteroatoms as ring atoms and the remaining ring atoms being carbon. The heteroatoms include oxygen, sulfur, and nitrogen, preferably 5-10 members, more preferably 5 or 6 members, such as thiophene, furanyl, pyridinyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc.

[0115] The term "hydroxyl-substituted C1-C6 alkyl" refers to straight-chain, branched, or cyclic C1-C6 alkyl groups substituted with one or more hydroxyl groups, including (S)-1-hydroxyisobutyl-2-yl, (R)-1-hydroxyisobutyl-2-yl, etc.

[0116] In the various groups of the present invention, C1-C6 can be independently C1, C2, C3, C4, C5 or C6, preferably C1-C3.

[0117] "Pharmaceutically acceptable salts" refer to drug molecules that form corresponding salts with organic acids, inorganic acids, or organic bases, such as hydrochloric acid, formic acid, trifluoroacetic acid, succinic acid, and methanesulfonates.

[0118] In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0119] Active ingredients

[0120] As used herein, “compound of the invention” or “active ingredient” refers to a compound of formula I, and also includes its pharmaceutically acceptable salts, tautomers, and stereoisomers:

[0121]

[0122] The definitions of X, X1, X2, X3, X4, Y, Y1, R1, and R2 are as described in this article.

[0123] Preferably, X, X1, X2, X3, X4, Y, Y1, R1, and R2 are independently and optionally the groups corresponding to those in the specific compounds of the present invention (especially the example compounds).

[0124] The compound represented by formula (I) may exist in different tautomer forms, all of which are included within the scope of this invention.

[0125] The term "tautomer" refers to structural isomers with different energies that interconvert via a low energy barrier. The reaction generally results in the movement of hydrogen atoms or protons, accompanied by the transformation of single bonds and adjacent double bonds.

[0126] The compounds of this invention can exist in specific geometric or stereoisomeric forms. All such compounds contemplated by this invention, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, non-corresponding isomers, racemates, and other mixtures, are within the scope of this invention. Additional asymmetric carbon atoms may be present in alkyl or other substituents. All such isomers and mixtures thereof are included within the scope of this invention.

[0127] The compounds represented by formula (I) may contain one or more asymmetric or chiral centers, and therefore may exist in different stereoisomer forms. The compounds of this invention include all stereoisomer forms, including but not limited to diastereomers, enantiomers, and transisomers, as well as mixtures thereof (such as racemates), all of which are included within the scope of this invention.

[0128] Unless otherwise stated, the term "enantiomer" refers to stereoisomers that are mirror images of each other.

[0129] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0130] A racemic mixture is a stereoisomer that is a mirror image of another, with opposite optical rotations that cancel each other out.

[0131] The compounds in the general formula and examples of this invention contain chiral centers; therefore, the single configuration or racemate of the compounds of this invention are also within the scope of this application.

[0132] This invention also aims to include the crystalline forms of the compounds, such as hydrates and solvates.

[0133] As used herein, the term "solvent complex" refers to a complex of the compound of the present invention coordinated with solvent molecules in a specific ratio.

[0134] As used herein, the term "hydrate" refers to a complex formed by the coordination of the compound of the present invention with water, such as a monohydrate.

[0135] "Pharmaceutically acceptable salts" refer to drug molecules that form corresponding salts with organic acids, inorganic acids, or organic bases, or inorganic bases. When the compounds of this invention contain a basic group, they can be prepared into pharmaceutically acceptable salts, including inorganic acid salts and organic acid salts. Suitable acids for salt formation 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, benzenesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

[0136] Some compounds of this invention may contain acidic fragments, including but not limited to carboxylic acids, which may form salts with various organic or inorganic bases. Typical base-formed salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and salts formed from organic bases.

[0137] This invention also includes any of the novel intermediates disclosed herein.

[0138] Uses, pharmaceutical compositions

[0139] In this application, "pharmaceutical composition" refers to a formulation of the compounds of the present invention with a medium generally accepted in the art for delivering bioactive compounds to mammals (e.g., humans). This medium includes pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate administration to the organism, thereby promoting the absorption of the active ingredient and the exertion of its bioactivity.

[0140] Typically, the pharmaceutical compositions of the present invention comprise the compound of formula I described herein, a pharmaceutically acceptable salt thereof, a tautomer, a stereoisomer, and a pharmaceutically acceptable carrier.

[0141] As used herein, the term "pharmaceutically acceptable" means 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, i.e., that the substance can be administered to an individual without causing an adverse biological reaction or interacting adversely with any component contained in the composition.

[0142] In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, flow aid, sweetener, diluent, preservative, dye / coloring agent, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that is permitted by the relevant government regulatory authority to be acceptable for human or animal use.

[0143] The compound of formula (I) can be used in combination with other known drugs for treating or improving similar symptoms. When administered in combination, the original drug's administration method and dosage can remain unchanged, while the compound of formula I is taken simultaneously or subsequently. When the compound of formula I is taken concurrently with one or more other drugs, a pharmaceutical composition containing one or more known drugs and the compound of formula I is preferred. Drug combination also includes taking the compound of formula I with 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 either drug alone.

[0144] Drugs or active ingredients that can be used in combination with compounds of general formula (I) include, but are not limited to, dexamethasone, rituximab, trastuzumab, PD-1 inhibitors, PD-L1 inhibitors, pemetrexed, topotecan, doxorubicin, bortezomib, gemcitabine, dacarbazine, clarithromycin, vincristine, cytarabine, azacitidine, prednisone, docetaxel, clofarabine injection, HDAC inhibitors, kinase-targeting inhibitors, androgen receptor inhibitors, androgen biosynthesis inhibitors, BTK inhibitors, BCL2 inhibitors, erythrocyte growth hormone, minocycline, elotuzumab, palbociclib, nivolumab, pembrolizumab, panobinostat, ublituximab, romidepsin, eltrombopag, CAR-T, and melphalan.

[0145] Typical formulations are prepared by mixing compounds of general 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 expandable polymers, hydrophilic or hydrophobic substances, gelatin, oils, solvents, and water. The specific carrier, diluent, or excipient used will depend on the manner and purpose of use of the compound according to 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. Generally, safe solvents are non-toxic aqueous solvents such as pharmaceutical water, as well as other non-toxic solvents that are soluble in 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, flow aids, processing aids, colorants, sweeteners, flavorings, seasonings or other known additives, so that the compound represented by formula (I) is manufactured or used in an acceptable form.

[0146] When the compounds of formula (I) of the present invention are used in combination with at least one other drug, the two or more drugs may be used separately or in combination, preferably in the form of a pharmaceutical composition. The compounds or pharmaceutical compositions of formula (I) of the present invention can be administered to the subject separately or together in any known oral, intravenous, rectal, vaginal, transdermal, or other local or systemic forms of administration.

[0147] These pharmaceutical compositions may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow aids, processing aids, colorants, sweeteners, flavorings, or other known additives to make or use the pharmaceutical composition in an acceptable form.

[0148] The preferred route of administration for the pharmaceutical products of this invention is oral administration. Solid dosage forms for oral administration may include capsules, tablets, powders, or granules. In solid dosage forms, the compounds or pharmaceutical compositions of this invention are 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, alginate, gelatin, polyvinylpyrrolidone, sucrose, gum arabic, etc.; humectants such as glycerin, etc.; disintegrants such as agar, calcium carbonate, potato or cassava starch, alginate, specific complex silicates, sodium carbonate, etc.; solution blocking agents 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. Similar solid compositions can also be used as fillers in soft and hard filled gelatin capsules, using lactose and high molecular weight polyethylene glycol as excipients.

[0149] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the compounds or compositions thereof of the present invention, the liquid dosage form may contain inert diluents 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-butanediol, dimethylformamide; oils (such as cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, etc.); glycerin; tetrahydrofurfuryl alcohol; fatty acid esters of polyethylene glycol and sorbitol; or mixtures of several of these substances.

[0150] In addition to these inert diluents, the composition may also include one or more excipients, such as wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, and fragrances.

[0151] In the case of suspensions, in addition to the compounds or combinations of the present invention, they may further contain carriers such as suspending agents, such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, dehydrated sorbitol ester, microcrystalline cellulose, aluminum hydroxide, bentonite, agar and astragalus gum, or mixtures of several of these substances.

[0152] Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing the compounds or combinations of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, or suppository wax, which are solid at normal room temperature and liquid at body temperature, and can melt in the rectum or vagina to release the active compound.

[0153] The compounds or pharmaceutical compositions of this invention can be administered in other topical dosage forms, including ointments, powders, sprays, and inhalers. The drug can be mixed under sterile conditions with pharmaceutically acceptable excipients, diluents, or carriers, and any desired preservatives, buffers, or propellants. Ophthalmic formulations, ophthalmic ointments, powders, and solutions are also intended to be covered within the scope of this invention.

[0154] As used herein, the terms “administration,” “application,” “dosage,” etc., refer to methods that deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral administration, duodenal administration, parenteral administration (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injection or infusion), local administration, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used with 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 a preferred embodiment, the compounds and compositions discussed herein are administered orally.

[0155] The terms "drug combination," "drug co-administration," "combined drug therapy," "administration of other treatments," and "administration of other therapeutic agents" used in this invention refer to drug therapy obtained by mixing or combining more than one active ingredient, including fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity or single dosage form. The term "non-fixed combination" refers to the simultaneous, combined, or sequential administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity at variable intervals. These also apply to cocktail therapies, such as the administration of three or more active ingredients.

[0156] The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to 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, 1-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0157] The compounds or pharmaceutical compositions of the present invention can be used to prepare treatments or preventative medications for CRL4. CRBN Drugs for diseases involving E3 ubiquitin ligases. This invention describes a drug involving CRL4. CRBN Diseases involving E3 ubiquitin ligase, without limitation, include tumors, central nervous system diseases, and immune diseases.

[0158] In a preferred embodiment, the diseases include, but are not limited to: cancer, pain (including but not limited to complex regional pain syndrome), skin diseases, immunodeficiency diseases, and damage and dysfunction of the central nervous system.

[0159] In another preferred embodiment, the cancers include (but are not limited to): skin cancers (such as melanoma), lymphatic system cancers, breast cancer, cervical cancer, uterine cancer, digestive tract cancers, lung cancer, ovarian cancer, prostate cancer, colon cancer, rectal cancer, oral cancer, brain tumors, head and neck cancer, pharyngeal cancer, testicular cancer, kidney cancer, pancreatic cancer, spleen cancer, liver cancer, bladder cancer, laryngeal cancer, and cancers related to AIDS. The compounds provided by this invention are also effective against hematologic malignancies and myeloma, such as for the treatment of multiple myeloma, lymphoma, and acute and chronic leukemia. The compounds provided by this invention can also be used for the prevention or treatment of primary and metastatic tumors.

[0160] The uses of the drug molecules of this invention include, but are not limited to, the diseases selected from: myelodysplastic syndromes, multiple myeloma, mantle cell lymphoma, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, myelofibrosis, Burkitt lymphoma, Hodgkin lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, ciliary body and chronic melanoma, iris melanoma, recurrent interocular melanoma, T-cell lymphoma, erythroid lymphoma, monocytic lymphoma. Monocytic and cytoplasmic 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, type I complex regional pain syndrome, malignant melanoma, radiculopathy, glioblastoma, glioma, malignant glioma, refractory plasmacytoma, extraocular melanoma, papillary and follicular thyroid carcinoma, breast cancer, prostate cancer, hepatocellular carcinoma, or primary macroglobulinemia.

[0161] The main advantages of this invention include:

[0162] 1. This invention provides a novel CRL4 with excellent activity. CRBN E3 ubiquitin ligase is a molecular gel-like degrader.

[0163] 2. The compounds of the present invention can specifically and efficiently degrade CK1α, while reducing the degradation of other proteins such as IKZF1 and GSPT1, thereby reducing the toxic side effects on normal cells.

[0164] 3. Compared with existing compounds, the pyridine heterocyclic substituted isoindoline compounds of the present invention have significantly lower hERG inhibitory activity, thereby reducing the risk of cardiotoxicity and improving the safety of the compounds in subsequent clinical use.

[0165] 4. The compounds of the present invention have excellent pharmacokinetic and pharmacodynamic properties, high bioavailability, and very good drug-like properties.

[0166] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

[0167] In all embodiments, 1The 1H NMR spectrometry was performed using a 400MHz and a 500MHz Bruker NMR spectrometer, with chemical shifts expressed as δ (ppm). Mass spectrometry was performed using UPLC-MS (ESI) mass spectrometry; 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 of benzophenone / sodium metal to remove oxygen, and anhydrous toluene and anhydrous dichloromethane were prepared by reflux drying of calcium chloride. Solvents used as mobile phases for column chromatography, 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 for reaction detection were from Sinopharm Chemical Reagent Co., Ltd. 200-300 mesh silica gel from Sinopharm Chemical Reagent Co., Ltd. was used for compound separation. The raw materials used in this invention can be obtained commercially, such as the main reagents purchased from Sinopharm Chemical Reagent Co., Ltd., or prepared by methods of inhibition in the art, or prepared according to the methods described in this invention.

[0168] Example

[0169] Synthesis of key intermediates:

[0170] Intermediate (1): 5-chloro-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine] hydrochloride

[0171]

[0172] Selenium dioxide (2.81 g, 25.34 mmol) was suspended in CH2Cl2 (120 mL). Tert-butyl hydroperoxide solution (13.9 mL, 0.101 mol, 70% in water) was added under ice bath conditions. After reacting for 30 min under ice bath conditions, compound 1.1 (10 g, 50.69 mmol) was dissolved in CH2Cl2 (20 mL) and added to the reaction system. The reaction was continued under ice bath conditions for 1 h, then slowly raised to room temperature. After 4 h at room temperature, TLC monitoring showed that the reaction was complete. The reaction was quenched with saturated sodium bisulfite solution, and the pH was adjusted to neutral with saturated sodium bicarbonate solution. Extraction was performed with EA, the organic phase was dried under rotary evaporation, and column chromatography was performed. A PE:EA ratio of 5:1 yielded 6.5 g of compound 1.2, with a yield of 60.12%. 1 H NMR (400MHz, CDCl3) δ5.01(s,1H),4.85(s,1H),4.08(s,1H),3.76(dd,J=12.8,4.0Hz,1H),3.52( d,J=26.0Hz,1H),3.16(d,J=30.3Hz,2H),2.41(dd,J=8.8,5.1Hz,1H),2.12(s,1H),1.44(s,9H).

[0173] Compound 1.2 (6.5 g, 30.48 mmol) was dissolved in benzene, and SOCl2 (2.67 mL, 36.88 mmol) was added. The reaction was carried out at 60 °C for 10 min. After the reaction was stopped, the mixture was transferred to an ice bath and quenched with saturated sodium bicarbonate solution. The mixture was extracted with EA, and the organic phase was dried and subjected to rotary evaporation for column chromatography. A PE:EA ratio of 10:1 yielded compound 1.3 as a yellow oily substance (3 g). 1 H NMR (400MHz, CDCl3) δ5.76(s,1H),4.02(s,2H),3.92(s,2H),3.53(s,2H),2.22(s,2H),1.47(s,9H).

[0174] Compound 1.4 (450 mg, 1.76 mmol) was added to a 100 mL round-bottom flask, along with compound 1.3 (408 mg, 1.76 mmol) and K₂CO₃ (487 mg, 3.52 mmol). 20 mL of ultradry DMF was used as the solvent. The reaction was carried out at 75 °C. After 4.5 h, LC-MS monitoring showed the reaction was complete. The mixture was diluted with saturated sodium chloride and extracted with EA. After washing away the DMF, the organic phase was dried and subjected to rotary cyclohexane chromatography (PE:EA ratio 5:1) to obtain 550 mg of compound 1.5, with a yield of 69.27%. 1 H NMR (400MHz, CDCl3) δ7.82(s,1H),7.75(s,1H),5.85(s,1H),4.56(s,2H),3.96(s,2H),3.56(t,J=5.6Hz,2H),2.23(s,2H),1.47(s,9H).

[0175] Compound 1.5 (550 mg, 1.22 mmol) was dissolved in 30 mL of toluene, and n-Bu3SnH (0.658 mL, 2.44 mmol) was added. The mixture was heated until the toluene began to boil, and then AIBN (200 mg, 1.22 mmol) was added. The reaction was carried out at 135 °C for 4 h. The reaction was monitored by TLC until complete, and the product spotted blue-violet with ninhydrin. Direct rotary cyclohexane column chromatography was performed. A PE:EA ratio of 4.0:1 to 3.5:1 yielded 350 mg of compound 1.6, with a yield of 88.30%. 1 H NMR (400MHz, CDCl3) δ7.92 (s, 1H), 7.07 (s, 1H), 4.48 (s, 2H), 4.06 (d, J = 11.5Hz, 2H), 2. 92(t,J=11.8Hz,2H),1.82(dd,J=11.4,4.3Hz,2H),1.74(d,J=13.4Hz,2H),1.48(s,9H).

[0176] Compound 1.6 (3500 mg, 1.08 mmol) was dissolved in 10 mL of CH2Cl2, and 4 M HCl in 1,4-Dioxane (2.5 mL, 10 mmol) was added. The reaction was carried out overnight at room temperature, and the reaction was monitored by TLC until complete. The solvent was evaporated, and the mixture was slurried with diethyl ether to obtain compound 1, which was used directly in the next step.

[0177] Intermediate (2): 5-(trifluoromethyl)-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine] hydrochloride

[0178]

[0179] Compound 2.1 (1 g, 6.13 mmol) was dissolved in 30 mL of dichloromethane, and DIPEA (1.28 mL, 7.36 mmol) was added. Bromomethyl methyl ether (0.5 mL, 6.13 mmol) was added under ice bath conditions. After 3 h, the reaction was monitored by TLC until complete. The mixture was diluted with saturated sodium chloride solution, extracted with CH2Cl2, and the organic phase was dried and subjected to rotary cyclohexane chromatography. A PE:EA ratio of 15:1 yielded 1.19 g of compound 2.2, with a yield of 93.69%. Compound 3.2 was dissolved in 20 mL of ultradry THF, purged with N2, and placed at -78 °C. 2.4 M n-butyllithium (2.87 mL, 6.89 mmol, 2.4 M in Hexane) was added. The reaction was carried out at -78 °C for 10 min. I2 (1.53 g, 6.03 mmol) was weighed, dissolved in 20 mL of ultradry THF, and added to the reaction mixture. The reaction was continued at -78°C for 5 hours. The mixture was then transferred to room temperature, and the reaction was quenched by adding sodium thiosulfate solution. Extraction was performed with diethyl ether, and the organic phase was dried and subjected to rotary evaporation for column chromatography. A PE:EA ratio of 17.5:1 yielded 900 mg of compound 2.3, with a yield of 47.04%. 1 HNMR (400MHz, CDCl3) δ8.38(s,1H),8.08(s,1H),5.36(s,2H),3.54(s,3H).

[0180] 900 mg of compound 2.3 was dissolved in CH₂Cl₂, and 4 M dioxane hydrochloride (5 mL, 20 mmol) was added. After reacting for 8 h, TLC monitoring showed that the reaction was complete. The mixture was neutralized to neutral with saturated sodium bicarbonate solution, extracted with CH₂Cl₂, and dried to 600 mg of compound 2.4, yielding 76.83%.

[0181] Compound 2.4 (350 mg, 1.21 mmol) was added to a flask, followed by compound 1.3 (309 mg, 1.33 mmol) and K₂CO₃ (335 mg, 2.42 mmol). 20 mL of ultradry DMF was used as the solvent. The reaction was carried out at 75 °C. After 4.5 h, LC-MS monitoring showed the reaction was complete. The mixture was diluted with saturated sodium chloride and extracted with EA. After washing away the DMF, the organic phase was dried and subjected to rotary cyclohexane chromatography (PE:EA ratio 6:1) to obtain 400 mg of compound 2.5, with a yield of 68.20%. 1 H NMR (400MHz, CDCl3) δ8.12(s,1H),8.08(s,1H),5.87(s,1H),4.67(s,2H),3.97(s,2H),3.57(t,J=5.6Hz,2H),2.24(s,2H),1.47(s,9H).

[0182] Compound 2.5 (400 mg, 0.826 mmol) was dissolved in 30 mL of toluene, and n-Bu3SnH (7.79 mL, 28.91 mmol) was added. The mixture was heated until the toluene began to boil, and then AIBN (1.63 g, 9.91 mmol) was added. The reaction was carried out at 135 °C for 4 h. The reaction was monitored by TLC until complete, and the product spotted blue-violet with ninhydrin. Direct rotary cyclohexane column chromatography was performed. A PE:EA ratio of 4.5:1 yielded 240 mg of compound 2.6, with a yield of 81.08%. 1H NMR (400MHz, CDCl3) δ8.25 (s, 1H), 7.45 (s, 1H), 4.56 (s, 2H), 4.11 (d, J = 9.8Hz, 2H ),2.91(t,J=12.2Hz,2H),1.91–1.85(m,2H),1.76(d,J=13.3Hz,2H),1.49(s,9H).

[0183] Compound 3.6 was dissolved in 10 mL of (240 mg, 0.670 mmol) solution, and 1.8 mL of 4 M dioxane hydrochloride (7.2 mmol) was added. The reaction was carried out overnight at room temperature, and the reaction was monitored by TLC until complete. The solvent was evaporated and the mixture was slurried with diethyl ether to give compound 2.

[0184] Intermediate (3): tert-butyl 7-chloro-5-(hydroxymethyl)-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0185]

[0186] The synthesis method is the same as that for intermediate 1. 1H NMR (400MHz, CDCl3) δ7.06 (s, 1H), 4.68 (s, 2H), 4.56 (s, 2H), 4.10 (dd, J = 15.7, 9.4Hz, 2H), 2. 91(t,J=12.5Hz,2H),1.85(td,J=12.7,11.7,4.5Hz,2H),1.76(d,J=13.2Hz,2H),1.48(s,9H).

[0187] Intermediate (4): tert-butyl 7-chloro-5-(difluoromethyl)-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0188]

[0189] The synthesis method is the same as that for intermediate 1. 1 H NMR (500MHz, CDCl3) δ7.38 (s, 1H), 6.56 (t, J = 55.5Hz, 1H), 4.63 (s, 2H), 4.12 (d, J = 6.9Hz, 2H), 2.90 (s, 2H), 1.89 (td, J = 12.7, 12.1, 4.5Hz, 2H), 1.79 (d, J = 13.4Hz, 2H), 1.49 (s, 9H).

[0190] Intermediate (5): tert-butyl 2H-spiro[furan[2,3-b]pyridine-3,4'-piperidine]-1'-carboxylate

[0191]

[0192] The synthesis method is the same as that for intermediate 1. 1 H NMR (500MHz, CDCl3) δ8.05 (dd, J=5.2, 1.5Hz, 1H), 7.42 (dd, J=7.2, 1.7Hz, 1H), 6.83 (dd, J=7.2, 5.2Hz,1H),4.43(s,2H),4.03(s,2H),2.97(s,2H),1.79(dd,J=37.2,11.9Hz,4H),1.48(s,9H).

[0193] Intermediate (6): 1'-benzyl-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]

[0194]

[0195] The synthesis method is the same as that for intermediate 1. 1H NMR (400MHz, CDCl3) δ8.20(d,J=4.7Hz,2H),7.36(d,J=4.5Hz,5H),7.14(d,J=4.9Hz,1H),4 .43(s,2H),3.58(s,2H),2.93(d,J=11.0Hz,2H),2.13–1.97(m,4H),1.76(d,J=12.6Hz,2H).

[0196] Intermediate (7): tert-butyl 5-fluoro-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0197]

[0198] The synthesis method is the same as that for intermediate 1. 1 H NMR (500MHz, CDCl3) δ7.69(d,J=1.5Hz,1H),6.70(d,J=3.0Hz,1H),4.51(s,2H),4.08(s,2H),2.94(s,2H),1.86–1.76(m,4H),1.50(s,9H).

[0199] Intermediate (8): tert-butyl 5-chloro-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0200]

[0201] The synthesis method is the same as that for intermediate 1. 1 H NMR (400MHz, CDCl3) δ7.92 (s, 1H), 7.07 (s, 1H), 4.48 (s, 2H), 4.06 (d, J = 11.5Hz, 2H), 2. 92(t,J=11.8Hz,2H),1.82(dd,J=11.4,4.3Hz,2H),1.74(d,J=13.4Hz,2H),1.48(s,9H).

[0202] Intermediate (9): tert-butyl 5-methyl-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0203]

[0204] The synthesis method is the same as that for intermediate 1. 1H NMR (400MHz, CDCl3) δ8.06 (s, 1H), 6.92 (s, 1H), 4.41 (s, 2H), 4.05 (d, J = 10.2Hz, 2H), 2.9 3(t,J=12.1Hz,2H),2.49(s,3H),1.86–1.79(m,2H),1.71(d,J=13.3Hz,2H),1.48(s,9H).

[0205] Intermediate (10): tert-butyl 5-(trifluoromethyl)-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0206]

[0207] The synthesis method is the same as that for intermediate 1. 1 H NMR (400MHz, CDCl3) δ8.25 (s, 1H), 7.45 (s, 1H), 4.56 (s, 2H), 4.11 (d, J = 9.8Hz, 2H), 2.9 1(t,J=12.2Hz,2H),1.88(dd,J=12.6,8.3Hz,2H),1.76(d,J=13.3Hz,2H),1.49(s,9H).

[0208] Intermediate (11): tert-butyl 5-(methyl-d3)-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0209]

[0210] The synthesis method is the same as that for intermediate 1. 1 H NMR (400MHz, CDCl3) δ8.07(s,1H),6.93(s,1H),4.42(s,2H),4.07(s,2H),2.92(d,J=11.6Hz,2H),1.84(s,2H),1.70(s,2H),1.49(s,9H).

[0211] Intermediate (12): 1'-benzyl-5,7-dichloro-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]

[0212]

[0213] The synthesis method is the same as that for intermediate 1. 1H NMR (400MHz, CDCl3) δ7.35–7.31(m,5H),7.05(s,1H),4.54(s,2H),3.54(s,2H),2.89 (d,J=11.9Hz,2H),2.05(t,J=11.6Hz,2H),1.99–1.92(m,2H),1.78(d,J=12.0Hz,2H).

[0214] Intermediate (13): tert-butyl 5-cyano-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine]-1'-carboxylate

[0215]

[0216] The synthesis method is the same as that for intermediate 1. 1 H NMR(400MHz, CDCl3)δ8.25(d,J=0.7Hz,1H),7.47(d,J=0.7Hz,1H),4.57(s,2H),4.13–4.0 6(m,2H),2.91(t,J=11.9Hz,2H),1.87–1.80(m,2H),1.75(d,J=13.3Hz,2H),1.48(s,9H).

[0217] Intermediate (14): (S)-3-(4-(5-bromopentyl)-1-oxoisoindololin-2-yl)piperidin-2,6-dione

[0218]

[0219] Compound 14.1 (10 g, 32.5 mmol) was dissolved in acetonitrile (100 mL), and (S)-3-aminopiperidine-2,6-dione hydrochloride (5.88 g, 35.7 mmol) and N,N-diisopropylethylamine (14.1 mL, 81 mmol) were added. The mixture was reacted overnight at 80°C. After the reaction was complete, the reaction solution was concentrated under reduced pressure, filtered, the filter cake was washed with methanol, the solid was collected, and dried to give 6.4 g of the product (S)-3-(4-bromo-1-oxoisoindoline-2-yl)piperidine-2,6-dione, a blue solid, with a yield of 61%. 1H NMR(500MHz,DMSO-d6)δ11.08(s,1H),7.87(dd,J=8.0,1.9Hz,1H),7.79(d,J=7 .5Hz,1H),7.52(t,J=7.7Hz,1H),5.18(dd,J=13.3,5.1Hz,1H),4.43(d,J=17.6H z,1H),4.27(d,J=17.6Hz,1H),2.94(ddd,J=17.3,13.7,5.4Hz,1H),2.62(ddd, J=17.3,4.6,2.3Hz,1H),2.51–2.42(m,1H),2.04(dtd,J=12.9,5.4,2.2Hz,1H). 13 C NMR (126MHz, DMSO) δ173.40,171.39,167.71,142.56,135.09,134.36,130.97,122.96,117.80,52.15,48.41,31.66,22.73.

[0220] The compound (S)-3-(4-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (6 g, 18.6 mmol) was dissolved in DMF (60 mL), and triethylamine (7.74 mL, 55.7 mmol), pentylo-4-yn-1-ol (4.32 mL, 46.4 mmol), cuprous iodide (0.530 g, 2.79 mmol), and bis(triphenylphosphine) palladium dichloride (0.651 g, 0.928 mmol) were added. The reaction was carried out overnight at 80°C under nitrogen protection. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was added with methanol. The mixture was stirred vigorously, filtered, and the filter cake was washed with methanol until white. The product (S)-3-(4-(5-hydroxypent-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione) was dried to give 5.5 g of the product (S)-3-(4-(5-hydroxypent-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione) as a grayish-white solid, with a yield of 91%. 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),7.72(d,J=7.5Hz,1H),7.64(d,J=7.6Hz,1H),7.53(t,J=7.6 Hz,1H),5.17(dd,J=13.4,5.1Hz,1H),4.61(t,J=5.1Hz,1H),4.46(d,J=17.8Hz,1H),4.32(d,J=17 .8Hz,1H),3.54(q,J=5.9Hz,2H),2.93(ddd,J=17.6,13.4,5.2Hz,1H),2.61(dt,J=17.1,3.3Hz,1H ),2.55–2.52(m,2H),2.50–2.42(m,1H),2.02(ddd,J=11.3,5.6,3.0Hz,1H),1.73(p,J=6.7Hz,2H). 13 C NMR (126MHz, DMSO) δ173.03,171.14,167.76,143.83,134.14,132.02,128.67,1 22.67,118.90,96.32,76.29,59.43,51.63,46.99,31.51,31.27,22.41,15.58.

[0221] Compound (S)-3-(4-(5-hydroxypent-1-yn-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (5.5 g, 16.8 mmol) was added to tetrahydrofuran (100 mL), and nickel catalyst (11 g) was added. The mixture was stirred vigorously at room temperature under a hydrogen atmosphere. After the reaction was complete, the reaction solution was filtered through diatomaceous earth, and the solid was washed with tetrahydrofuran. The filtrate was concentrated under reduced pressure to give 4.5 g of product (S)-3-(4-(5-hydroxypentyl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione, a white solid, in 81% yield. 1H NMR(500MHz,DMSO-d6)δ11.05(s,1H),7.57(q,J=4.1Hz,1H),7.50–7.44(m,2H),5.16(dd,J= 13.3,5.1Hz,1H),4.53–4.37(m,2H),4.31(d,J=17.1Hz,1H),3.39(t,J=6.5Hz,2H),2.94(dd d,J=18.1,13.6,5.4Hz,1H),2.67–2.58(m,3H),2.45(qd,J=13.0,4.3Hz,1H),2.02(dq,J=12 .0,5.6,4.9Hz,1H),1.60(p,J=7.6Hz,2H),1.46(p,J=6.7Hz,2H),1.34(h,J=7.3,6.4Hz,2H). 13 C NMR (126MHz, DMSO) δ173.06,171.21,168.47,140.58,137.61,131.59,128. 35,120.66,60.68,51.58,46.28,32.37,31.38,31.28,29.34,25.56,22.57.

[0222] (S)-3-(4-(5-hydroxypentyl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (4.5 g, 13.6 mol) was added to dry tetrahydrofuran (50 mL), followed by triphenylphosphine (5.72 g, 21.8 mmol) and carbon tetrabromide (7.23 g, 21.8 mmol). The mixture was stirred at room temperature. After the reaction was complete, the solution was concentrated under reduced pressure, ethyl acetate was added, and the mixture was stirred vigorously. The mixture was filtered, the filter cake was washed with methanol, and dried to give 3.9 g of the product as a white solid, with a yield of 72%. 1 H NMR(500MHz,DMSO-d6)δ11.02(s,1H),7.60–7.55(m,1H),7.48–7.42(m,2H),5.15(dd,J =13.3,5.2Hz,1H),4.48(d,J=17.1Hz,1H),4.32(d,J=17.1Hz,1H),3.54(t,J=6.7Hz,2H ),2.94(ddd,J=18.2,13.7,5.4Hz,1H),2.68–2.57(m,3H),2.44(qd,J=13.1,4.3Hz,1H) ,2.06–1.98(m,1H),1.84(p,J=6.9Hz,2H),1.63(p,J=7.5Hz,2H),1.44(p,J=7.6Hz,2H). 13C NMR (126MHz, DMSO) δ172.94,171.09,168.38,140.54,137.31,131.57,131.49,1 28.29,120.67,51.54,46.24,35.15,32.03,31.23,31.11,28.36,27.51,22.55.

[0223] Intermediate (15): 3-(4-(5-bromopentyl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione

[0224]

[0225] Using compound 15.1 as the starting material, the synthetic route is the same as that of intermediate 15. 1 H NMR (500MHz, DMSO) δ11.01(s,1H),7.62(dd,J=11.8,7.3Hz,1H),7.56(dd,J=6.5,4.0Hz,1H),7.48– 7.43(m,1H),5.14(dd,J=13.4,5.2Hz,1H),4.47(d,J=17.1Hz,1H),4.31(d,J=17.1Hz,1H),3.54(t, J=6.6Hz,2H),2.98–2.87(m,1H),2.63(dd,J=22.8,14.8Hz,3H),2.43(ddd,J=26.4,13.4,4.3Hz,1H ),2.06–1.97(m,1H),1.94–1.76(m,2H),1.63(dt,J=15.3,7.6Hz,2H),1.44(dt,J=14.8,7.5Hz,2H).

[0226] Intermediate (16): 3-(4-(5-bromopentyl)-6-fluoro-1-oxoisoindololin-2-yl)piperidine-2,6-dione

[0227]

[0228] 25 g (117 mmol) of methyl 5-fluoro-2-methyl-3-nitrobenzene was dissolved in methanol (250 mL), and 2.5 g of 10% Pd / C was added. The mixture was stirred at room temperature for 8 hours under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give 21.1 g of methyl 3-amino-5-fluoro-2-methylbenzoate, with a yield of 98%. 1 H NMR (500MHz, DMSO-d6) δ6.63–6.56(m,2H),5.45(s,2H),3.78(s,3H),2.13(s,3H). 13C NMR (126MHz, DMSO-d6) δ167.79 (d, J = 3.3Hz), 160.41 (d, J = 238.2Hz), 149.67 (d, J = 11.1Hz), 132.29 (d, J =9.5Hz),117.21(d,J=2.3Hz),102.79(d,J=24.0Hz),102.66(d,J=23.6Hz),52.04,13.43.ESI-MS[M+H] + m / z = 184.13.

[0229] 21.1 g (115 mmol) of methyl 3-amino-5-fluoro-2-methylbenzoate was dissolved in dry acetonitrile (200 mL), and cuprous bromide (21.5 g (150 mmol), copper bromide (1.41 g (6.3 mmol), and tert-butyl nitrite (23.8 g (231 mmol)) were added. The mixture was reacted at 30°C for 18 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The residue was diluted with ethyl acetate, washed with water, and washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography to give 20.8 g of methyl 3-bromo-5-fluoro-2-methylbenzoate as a colorless oil, with a yield of 73%. 1 H NMR (500MHz, DMSO-d6) δ7.80 (dd, J=8.1, 2.8Hz, 1H), 7.54 (dd, J=8.9, 2.8Hz, 1H), 3.85 (s, 3H), 2.45 (s, 3H). 13 C NMR (126MHz, DMSO-d6) δ166.14(d,J=2.8Hz), 159.23(d,J=247.5Hz), 133.51(d,J=5.3 Hz), 126.33 (d, J = 9.3Hz), 122.78 (d, J = 24.0Hz), 115.84 (d, J = 22.9Hz), 52.71, 19.41.

[0230] 20.8 g (84 mmol) of methyl 3-bromo-5-fluoro-2-methylbenzoate was dissolved in 200 mL of 1,2-dichloroethane. N-bromosuccinimide (15.7 g, 88 mmol) and benzoyl peroxide (2.04 g, 8.4 mmol) were added, and the mixture was reacted overnight at 80°C. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The residue was diluted with ethyl acetate, washed with water, and then washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 26.6 g of methyl 3-bromo-2-(bromomethyl)-5-fluorobenzoate as an oil, with a yield of 97%. 1H NMR (500MHz, DMSO-d6) δ7.95 (dd, J=8.0, 2.8Hz, 1H), 7.70 (dd, J=8.9, 2.8Hz, 1H), 4.98 (s, 2H), 3.89 (s, 3H).

[0231] Methyl 3-bromo-2-(bromomethyl)-5-fluorobenzoate (20 g, 61.4 mmol) was dissolved in acetonitrile (200 mL), and 3-amino-2,6-piperidinidone hydrochloride (12 g, 73.6 mmol) and N,N-diisopropylethylamine (32 mL, 184 mmol) were added. The reaction was carried out overnight at 80°C. After the reaction was complete, the reaction solution was concentrated under reduced pressure, filtered, the filter cake was washed with methanol, the solid was collected, and dried to give 14.3 g of the product 3-(4-bromo-6-fluoro-1-oxoisoindoline-2-yl)piperidin-2,6-dione as a blue solid, with a yield of 68%. 1 HNMR(500MHz,DMSO-d6)δ11.02(s,1H),7.89(dd,J=8.9,2.3Hz,1H),7.63(dd,J=7.4,2.3Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.40(dd,J=1 7.5,1.3Hz,1H),4.25(dd,J=17.5,1.4Hz,1H),2.91(ddd,J=17.3,13.7,5.4Hz,1H),2.64–2.56(m,1H),2.49–2.40(m,1H),2.05–1.98(m,1H). 13 C NMR (126MHz, DMSO-d6) δ172.85, 170.73, 166.41, 162.21 (d, J = 249.5Hz), 138.25, 135.05 (d, J = 8.6Hz), 122 .34(d,J=26.7Hz),117.79(d,J=10.0Hz),109.63(d,J=23.4Hz),51.96,47.73,31.16,22.22.ESI-MS[M+H] + m / z = 340.98, 342.92.

[0232] 20 g (58.6 mmol) of compound 3-(4-bromo-6-fluoro-1-oxoisoindololin-2-yl)piperidine-2,6-dione was dissolved in DMF (200 mL), and triethylamine (24.5 mL, 176 mmol), penta-4-yn-1-ol (13.6 mL, 146 mmol), cuprous iodide (1.67 g, 8.79 mmol), and palladium dichloride bis(triphenylphosphine) (2.06 g, 2.93 mmol) were added. The reaction was carried out overnight at 80 °C under nitrogen protection. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was added with methanol. The mixture was stirred vigorously, filtered, and the filter cake was washed with methanol until white. The product was dried to give 18.7 g of white solid, with a yield of 93%. 1 H NMR(500MHz,DMSO-d6)δ11.01(s,1H),7.54(s,1H),7.52(s,1H),5.14(dd,J=13.3,5.1 Hz,1H),4.56(t,J=5.1Hz,1H),4.43(d,J=17.7Hz,1H),4.29(d,J=17.6Hz,1H),3.52(q ,J=5.9Hz,2H),2.91(ddd,J=17.2,13.6,5.5Hz,1H),2.63–2.56(m,1H),2.53(t,J=7.1 Hz,2H),2.49–2.39(m,1H),2.02(ddt,J=12.5,7.6,3.8Hz,1H),1.71(p,J=6.7Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ172.80, 170.80, 166.71, 161.95 (d, J = 244.2Hz), 139.73, 133.81 (d, J = 9.5Hz), 121.05 (d, J = 25.0Hz) ,120.51(d,J=10.6Hz),109.65(d,J=23.8Hz),97.64,75.26,59.30,51.75,46.66,31.25,31.11,22.21,15.46.ESI-MS[M+H] + m / z = 345.17.

[0233] Compound 3-(6-fluoro-4-(5-hydroxypent-1-yn-1-yl)-1-oxoisoindololin-2-yl)piperidin-2,6-dione (18.7 g, 54.3 mmol) was added to tetrahydrofuran (400 mL), and a nickel catalyst (20 g) was added. The mixture was stirred vigorously at room temperature under a hydrogen atmosphere. After the reaction was complete, the reaction solution was filtered through diatomaceous earth, and the solid was washed with tetrahydrofuran. The filtrate was concentrated under reduced pressure to give 17.4 g of the product as a white solid, with a yield of 92%. 1H NMR(500MHz,DMSO-d6)δ11.00(s,1H),7.35(s,1H),7.33(s,1H),5.13(dd,J=13.3 ,5.2Hz,1H),4.45(d,J=17.4Hz,1H),4.29(d,J=17.1Hz,1H),3.39(t,J=6.3Hz,2H) ,2.92(ddd,J=18.4,13.4,5.3Hz,1H),2.67–2.57(m,3H),2.48–2.36(m,1H),2.06 –1.98(m,1H),1.61(p,J=7.7Hz,2H),1.45(p,J=6.7Hz,2H),1.34(p,J=7.7Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ172.86, 170.90, 167.37, 162.37 (d, J = 243.7Hz), 140.37, 136.47, 133.36 (d, J = 9.2Hz), 11 8.60(d,J=23.4Hz),106.99(d,J=23.6Hz),60.58,51.73,46.04,32.23,31.14,28.90,25.39,22.41.ESI-MS[M+H] + m / z = 349.13.

[0234] Compound 3-(6-fluoro-4-(5-hydroxypentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione (13 g, 37.3 mol) was added to dry tetrahydrofuran (200 mL), followed by triphenylphosphine (19.6 g, 74.6 mmol) and carbon tetrabromide (24.8 g, 74.6 mmol). The mixture was stirred at room temperature. After the reaction was complete, the reaction solution was concentrated under reduced pressure, diluted with dichloromethane, and then ethyl acetate was added. The mixture was stirred vigorously, and a solid precipitated. The solid was filtered, washed with methanol, and dried to give 8.3 g of the product as a white solid, with a yield of 54%. 1H NMR(500MHz,DMSO-d6)δ11.01(s,1H),7.39–7.31(m,2H),5.13(dd,J=13.3,5.1Hz,1H),4 .45(d,J=17.2Hz,1H),4.30(d,J=17.3Hz,1H),3.54(t,J=6.7Hz,2H),2.92(ddd,J=17.3,1 3.7,5.4Hz,1H),2.69–2.63(m,2H),2.63–2.57(m,1H),2.42(qd,J=13.3,4.5Hz,1H),2.0 2(dtt,J=14.1,5.8,2.9Hz,1H),1.89–1.80(m,2H),1.69–1.58(m,2H),1.49–1.39(m,2H). 13 C NMR (126MHz, DMSO-d6) δ172.86, 170.89, 167.37, 162.39 (d, J = 243.9Hz), 140.16 (d, J = 8.0Hz), 136.49, 133.39 (d, J = 8.9Hz) ,118.61(d,J=22.9Hz),107.06(d,J=23.4Hz),51.73,46.05,35.08,31.95,31.15,30.89,27.98,27.37,22.45.ESI-MS[M+H] + m / z = 411.02, 413.04.

[0235] Intermediate (17): (S)-3-(4-(5-bromopentyl)-6-fluoro-1-oxoisoindololin-2-yl)piperidin-2,6-dione

[0236]

[0237] Methyl 3-bromo-2-(bromomethyl)-5-fluorobenzoate (9 g, 27.6 mmol) was dissolved in acetonitrile (90 mL), and (S)-3-aminopiperidine-2,6-dione hydrochloride (5.45 g, 33.1 mmol) and N,N-diisopropylethylamine (12 mL, 69.0 mmol) were added. The mixture was reacted overnight at 80°C. After the reaction was complete, the reaction solution was concentrated under reduced pressure, filtered, the filter cake was washed with methanol, the solid was collected, and dried to give 4.5 g of (S)-3-(4-bromo-6-fluoro-1-oxoisoindololin-2-yl)piperidine-2,6-dione, a pale purple solid, with a yield of 48%. 1H NMR (500MHz, DMSO-d6) δ11.07(s,1H),7.92(dd,J=8.9,2.2Hz,1H),7.65(dd,J=7.3,2.2Hz,1H),5.17(dd,J=13.3,5.1Hz,1H),4.41(d,J=17.5Hz ,1H),4.25(d,J=17.4Hz,1H),2.93(ddd,J=17.2,13.7,5.4Hz,1H),2.60 (ddd,J=17.3,4.5,2.2Hz,1H),2.50–2.42(m,1H),2.11–1.99(m,1H).13C NMR (126MHz, DMSO-d6) δ172.97, 170.84, 166.48, 162.25 (d, J = 249.3Hz), 138.31, 135.10, 122 .40(d,J=26.9Hz),117.87(d,J=10.0Hz),109.70(d,J=23.6Hz),51.94,47.73,31.21,22.25.

[0238] The compound (S)-3-(4-bromo-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (4.5 g, 13.2 mmol) was dissolved in DMF (60 mL), and triethylamine (9.2 mL, 65.9 mmol), pentan-4-yn-1-ol (3.07 mL, 32.9 mmol), cuprous iodide (0.376 g, 1.98 mmol), and palladium dichloride bis(triphenylphosphine) (0.555 g, 0.791 mmol) were added. The reaction was carried out overnight at 80°C under nitrogen protection. After the reaction was complete, the reaction solution was concentrated under reduced pressure. Methanol was added to the residue, and the mixture was stirred vigorously. The mixture was filtered, and the filter cake was washed with methanol until white. After drying, 3.9 g of the product (S)-3-(6-fluoro-4-(5-hydroxypent-1-yn-1-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione was obtained as a grayish-white solid, with a yield of 86%. ¹H NMR (500 MHz, DMSO-d6) δ 11.04 (s, 1H), 7.56–7.50 (m, 2H), 5.16 (dd, J = 13.3, 5.1 Hz, 1H), 4.60 (t, J = 5.0 Hz, 1H), 4.44 (d, J = 17.6 Hz, 1H), 4.31 (d, J = 17.6 Hz, 1H), 3.54 (dq, J = 5.9, 3.2 Hz, 2H), 2. 93(ddd,J=17.0,13.5,5.2Hz,1H),2.62(ddd,J=17.3,4.6,2.1Hz,1H),2.55(t,J=7.1Hz,2H),2 .47(td,J=13.2,4.5Hz,1H),2.04(ddq,J=10.4,5.4,3.2,2.6Hz,1H),1.73(p,J=6.7Hz,2H).13C NMR (126MHz, DMSO-d6) δ172.91, 170.90, 166.80 (d, J = 3.7Hz), 162.03 (d, J = 245.4Hz), 139.80, 133.88 (d, J = 9.2Hz), 121.12 ( d,J=25.3Hz),120.59(d,J=10.2Hz),109.73(d,J=23.6Hz),97.71,75.35,59.40,51.84,46.74,31.33,31.20,22.31,15.55.

[0239] Compound (S)-3-(6-fluoro-4-(5-hydroxypent-1-yn-1-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (3.9 g, 11.3 mmol) was added to tetrahydrofuran (100 mL), and nickel catalyst (8 g) was added. The mixture was stirred vigorously at room temperature under a hydrogen atmosphere. After the reaction was complete, the reaction solution was filtered through diatomaceous earth, and the solid was washed with tetrahydrofuran. The filtrate was concentrated under reduced pressure to give 3.3 g of product (S)-3-(6-fluoro-4-(5-hydroxypentyl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione, a white solid, in 83% yield. NMR(500MHz,DMSO-d6)δ11.03(s,1H),7.36(s,1H),7.34(s,1H),5.14(dd,J=13.3,5.1Hz,1H) ,4.46(d,J=17.1Hz,1H),4.38(t,J=5.1Hz,1H),4.30(d,J=17.1Hz,1H),3.39(q,J=6.3,4.9Hz, 2H),2.93(ddd,J=17.3,13.7,5.4Hz,1H),2.68–2.57(m,3H),2.44(qd,J=13.2,4.4Hz,1H),2. 02(dtd,J=12.7,5.3,2.2Hz,1H),1.66–1.55(m,2H),1.51–1.41(m,2H),1.39–1.29(m,2H).13C NMR (126MHz, DMSO-d6) δ172.93, 170.97, 167.43, 162.41 (d, J = 244.1Hz), 140.43 (d, J = 8.1Hz), 136.50, 133.39 (d, J=9.0Hz),118.64(d,J=23.3Hz),107.04(d,J=23.4Hz),60.62,51.75,46.07,32.27,31.18,28.95,25.43,22.45.

[0240] Compound (S)-3-(6-fluoro-4-(5-hydroxypentyl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (1.5 g, 4.31 mol) was added to dry tetrahydrofuran (30 mL), followed by triphenylphosphine (1.69 g, 6.46 mmol) and carbon tetrabromide (2.14 g, 6.46 mmol). The mixture was stirred at room temperature. After the reaction was complete, the reaction solution was concentrated under reduced pressure. The residue was separated by silica gel column chromatography to give 1.3 g of the product, a white solid, in 73% yield. 1H NMR(500MHz,DMSO-d6)δ11.03(s,1H),7.40–7.29(m,2H),5.15(dd,J=13.4,5.1Hz, 1H),4.46(d,J=17.1Hz,1H),4.31(d,J=17.1Hz,1H),3.55(t,J=6.7Hz,2H),2.93(d dd,J=18.0,13.7,5.4Hz,1H),2.69–2.57(m,3H),2.43(qd,J=13.3,4.5Hz,1H),2.0 7–1.98(m,1H),1.85(p,J=6.9Hz,2H),1.64(p,J=7.5Hz,2H),1.44(p,J=7.7Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ172.91, 170.93, 167.39, 162.40 (d, J = 244.2Hz), 140.17 (d, J = 8.1Hz), 136.51, 133.40 (d, J = 9 .0Hz), 118.63 (d, J = 23.1Hz), 107.09 (d, J = 23.2Hz), 51.73, 46.06, 35.13, 31.97, 31.17, 30.91, 28.00, 27.40, 22.47.

[0241] Synthesis of the compounds in the examples:

[0242] Example 1: 3-(4-(5-(2H-spirocyclic [furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0243]

[0244] Intermediate 3-(4-(5-bromopentyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (60 mg, 0.15 mmol) and 2H-spiro[furan[2,3-c]pyridine-3,4'-piperidine] hydrochloride (34 mg, 0.15 mmol) were dissolved in 2 mL of DMF. Triethylamine (208 μL, 1.5 mmol) was added, and the mixture was reacted overnight at room temperature. After the reaction was completed by LC-Mass monitoring, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed four times with saturated sodium chloride solution, dried, filtered, concentrated, and purified by HPLC to give 29 mg of white solid, with a yield of 38%. 1 H NMR (500MHz, DMSO-d6) δ11.01(s,1H),8.13–8.11(m,2H),7.58(dd,J=5.4,3.2Hz,1H),7.48–7.46(m,2H),7.34(d,J =4.2Hz,1H),5.16–5.12(m,1H),4.47(d,J=17.1Hz,1H),4.42(s,2H),4.31(d,J=17.2Hz,1H),2.95–2.90(m,1H),2. 81(d,J=11.4Hz,2H),2.68–2.64(m,2H),2.59(s,1H),2.46–2.42(m,1H),2.31–2.27(m,2H),2.04–2.00(m,1H),1.9 5(t,J=11.2Hz,2H),1.85(t,J=10.7Hz,2H),1.66–1.61(m,4H),1.51–1.47(m,2H),1.36–1.32(m,2H).ESI-MS[M+H] + m / z = 503.44.

[0245] Example 2: 3-(4-(5-(2H-spirocyclic [furan[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0246]

[0247] The synthesis method and feed ratio were the same as in Example 1, yielding 21 mg of white solid, with a yield of 29%. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),8.06(t,J=5.6Hz,1H),7.59(t,J=4.3Hz,1H),7.52–7.4 5(m,2H),7.28–7.15(m,2H),5.16(dd,J=13.3,5.0Hz,1H),4.63–4.39(m,3H),4.31(d,J=17.0 Hz,1H),3.19–3.00(m,4H),2.98–2.89(m,1H),2.76–2.59(m,3H),2.43–2.35(m,2H),2.14(dd ,J=25.6,12.2Hz,2H),2.09–1.91(m,4H),1.75–1.63(m,4H),1.44–1.33(m,2H).ESI-MS[M+H] + m / z = 503.65.

[0248] Example 3: 3-(4-(5-(5-fluoro-2H-spirocyclic [furan[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0249]

[0250] The synthesis method and feed ratio were the same as in Example 1, yielding 30 mg of white solid, with a yield of 38%. 1 H NMR (500MHz, DMSO-d6) δ11.02(s,1H),7.60–7.57(m,1H),7.49–7.43(m,3H),7.02(t,J=7.7Hz,1H),6.97(dd,J=8 .6,1.5Hz,1H),5.15(dd,J=13.3,5.0Hz,1H),4.65(s,1H),4.48(d,J=11.5Hz,1H),4.31(d,J=17.1Hz,1H),3.48( d,J=14.9Hz,2H),3.13–2.96(m,4H),2.96–2.90(m,1H),2.68(t,J=7.6Hz,2H),2.63(s,1H),2.41(dd,J=12.9,3. 9Hz,1H),2.13–2.09(m,1H),2.08–1.94(m,4H),1.68(dd,J=16.4,8.6Hz,4H),1.36(d,J=7.3Hz,2H).ESI-MS[M+H] + m / z = 521.47.

[0251] Example 4: 3-(4-(5-(5-chloro-2H-spirocyclic [furan[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0252]

[0253] The synthesis method and feed ratio were the same as in Example 1, yielding 18 mg of white solid, with a yield of 22%. 1 H NMR (500MHz, DMSO-d6) δ11.02(s,1H),7.58(d,J=4.3Hz,1H),7.48(d,J=4.4Hz,2H),7.35(d,J=8.6Hz,1H),7.29(d ,J=8.5Hz,1H),5.15(dd,J=13.3,4.9Hz,1H),4.65(s,2H),4.49–4.46(m,1H),4.31(d,J=17.1Hz,1H),3.49(s,2H) ,3.05(dd,J=13.2,9.2Hz,4H),2.94(s,1H),2.68(t,J=7.5Hz,2H),2.63(s,1H),2.41(dd,J=13.1,4.2Hz,1H),2.1 2(t,J=12.0Hz,2H),2.00(dd,J=22.4,10.0Hz,3H),1.74–1.64(m,4H),1.36(dd,J=15.0,7.6Hz,2H).ESI-MS[M+H] + m / z = 537.45.

[0254] Example 5: 3-(1-oxo-4-(5-(5-(trifluoromethyl)-2H-spiro[furan[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)isoindoline-2-yl)piperidin-2,6-dione

[0255]

[0256] The synthesis method and feed ratio were the same as in Example 1, yielding 20 mg of white solid, with a yield of 23%. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),7.78–7.70(m,1H),7.60(d,J=3.9Hz,1H),7.47(dt,J=8.6,4.5Hz ,3H),5.21–5.13(m,1H),4.76(d,J=4.0Hz,2H),4.49(dd,J=17.1,3.9Hz,1H),4.32(dd,J=17.1,4.1Hz, 1H),3.55(t,J=27.3Hz,3H),3.09(s,3H),2.94(t,J=15.4Hz,1H),2.67(d,J=21.7Hz,3H),2.45–2.37(m ,1H),2.20(t,J=14.1Hz,2H),2.04(d,J=12.8Hz,3H),1.70(d,J=25.8Hz,4H),1.38(s,2H).ESI-MS[M+H] + m / z = 571.66.

[0257] Example 6: 3-(4-(5-(2H-spirocyclic [furan[2,3-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0258]

[0259] The synthesis method and feed ratio were the same as in Example 1, yielding 16 mg of white solid at 21%. 1 H NMR (400MHz, DMSO-d6) δ11.02(s,1H),9.66(s,1H),8.43(s,1H),8.38(d,J=5.2Hz,1H),7.58(dd,J=8.4, 4.3Hz,1H),7.54(d,J=5.1Hz,1H),7.48(d,J=4.0Hz,2H),5.15(dd,J=13.3,5.1Hz,1H),3.56(d,J=12.1Hz ,2H),3.38(s,1H),3.31(s,1H),3.16–2.87(m,5H),2.67(dd,J=15.6,8.3Hz,3H),2.46–2.34(m,1H),2.16 (t,J=12.0Hz,2H),2.02(d,J=14.2Hz,3H),1.79–1.60(m,4H),1.37(dd,J=14.4,7.1Hz,2H).ESI-MS[M+H] + m / z = 503.40

[0260] Example 7: 3-(4-(5-(7-chloro-5-(hydroxymethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0261]

[0262] The synthesis method and feed ratio were the same as in Example 1, yielding 19 mg of white solid, with a yield of 22%. 1 H NMR(500MHz,DMSO-d6)δ11.01(s,1H),7.57(dd,J=5.3,3.3Hz,1H),7.49–7.44(m,2H),7.35(d,J=3.4Hz,1H),5 .45(t,J=6.0Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.54(s,2H),4.50–4.28(m,4H),2.93(ddd,J=17.3,13.7,5. 4Hz,1H),2.80(s,2H),2.66(t,J=7.7Hz,2H),2.64–2.57(m,1H),2.44(td,J=13.1,4.4Hz,1H),2.29(s,2H),2. 05–1.90(m,3H),1.82(s,2H),1.72(s,2H),1.63(p,J=7.7Hz,2H),1.50(s,2H),1.37–1.32(m,2H).ESI-MS[M+H] + m / z = 567.83.

[0263] Example 8: 3-(4-(5-(7-chloro-5-(difluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0264]

[0265] The synthesis method and feed ratio were the same as in Example 1, yielding 20 mg of white solid, with a yield of 22%. 1H NMR (500MHz, DMSO-d6) δ11.02(s,1H),7.73(s,1H),7.57(dd,J=5.1,3.4Hz,1H),7.50–7.42(m,2H),6.87(t,J=55.0Hz,1H),5.14 (dd,J=13.4,5.1Hz,1H),4.64(s,2H),4.39(dd,J=81.9,17.1Hz,2H),2.93(ddd,J=17.3,13.7,5.4Hz,1H),2.82(dd,J=8.7,5.3H z,2H),2.65(t,J=7.7Hz,2H),2.63–2.56(m,1H),2.43(qd,J=13.2,4.5Hz,1H),2.28(t,J=7.3Hz,2H),2.01(ddd,J=9.7,5.3,2.6 Hz,1H),1.98–1.85(m,4H),1.78–1.69(m,2H),1.62(p,J=7.6Hz,2H),1.48(p,J=7.3Hz,2H),1.33(p,J=7.7Hz,2H).ESI-MS[M+H] + m / z = 587.31.

[0266] Example 9: 3-(4-(5-(5-fluoro-2H-spirocyclic [furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0267]

[0268] The synthesis method and feed ratio were the same as in Example 1, yielding 38 mg of white solid, with a yield of 49%. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),7.67(s,1H),7.60–7.56(m,1H),7.49–7.45(m,2H),7.18(s,1H),5.15(dd ,J=13.3,5.1Hz,1H),4.48(d,J=19.7Hz,3H),4.31(d,J=17.1Hz,1H),2.97–2.90(m,1H),2.83(s,2H),2.69–2.6 4(m,2H),2.61(d,J=16.7Hz,1H),2.47–2.40(m,1H),2.30(s,2H),2.02(dd,J=6.6,4.3Hz,1H),1.90(d,J=10.1H z,4H),1.73–1.66(m,2H),1.63(dd,J=15.1,7.7Hz,2H),1.50(s,2H),1.35(dd,J=14.8,7.7Hz,2H).ESI-MS[M+H] + m / z = 521.43.

[0269] Example 10: 3-(4-(5-(5-(difluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0270]

[0271] The synthesis method and feed ratio were the same as in Example 1, yielding 27 mg of white solid, with a yield of 33%. 1H NMR (500MHz, DMSO-d6) δ11.02(s,1H),8.21(s,1H),7.65(s,1H),7.57(dt,J=7.7,3.9Hz,1H),7.48–7.44(m,2H),6.85(t,J=55.3H z,1H),5.14(dd,J=13.3,5.1Hz,1H),4.53(s,2H),4.39(dd,J=82.5,17.2Hz,2H),2.92(ddd,J=17.4,13.7,5.4Hz,1H),2.83(s,2H ),2.65(t,J=7.8Hz,2H),2.60(dt,J=17.2,3.3Hz,1H),2.43(qd,J=13.2,4.4Hz,1H),2.30(s,2H),2.01(dtd,J=10.7,5.5,2.7Hz, 2H),1.91(d,J=12.3Hz,3H),1.68(d,J=12.3Hz,2H),1.62(q,J=7.8Hz,2H),1.50(t,J=7.5Hz,2H),1.39–1.29(m,2H).ESI-MS[M+H] + m / z = 553.40.

[0272] Example 11: 3-(4-(5-(5-(difluoromethyl)-2H-spiro[furan[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0273]

[0274] The synthesis method and feed ratio were the same as in Example 1, yielding 31 mg of white solid, with a yield of 37%. 1H NMR (500MHz, DMSO-d6) δ11.01(s,1H),7.57(dd,J=5.6,3.0Hz,1H),7.50–7.46(m,2H),7.45(d,J=8.2Hz,1H),7.32(d,J=8.3Hz ,1H),6.89(t,J=55.2Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.56(s,2H),4.51–4.27(m,2H),2.92(ddd,J=17.3,13.6,5.5Hz,3H ),2.70–2.62(m,3H),2.63–2.56(m,1H),2.47–2.39(m,1H),2.36(p,J=1.9Hz,2H),2.01(dtd,J=12.7,5.3,2.2Hz,2H),1.93(t ,J=11.1Hz,2H),1.69(d,J=13.0Hz,2H),1.63(q,J=7.8Hz,2H),1.51(q,J=8.6,7.5Hz,2H),1.35(p,J=7.8Hz,2H).ESI-MS[M+H] + m / z = 553.32.

[0275] Example 12: 3-(4-(5-(5-chloro-2H-spirocyclic [furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0276]

[0277] The synthesis method and feed ratio were the same as in Example 1, yielding 38 mg of white solid, with a yield of 47%. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),7.93(s,1H),7.58–7.56(m,1H),7.48–7.46(m,3H),5.16–5.13(m,1H ),4.49(s,2H),4.45(s,1H),4.32–4.29(m,1H),2.93(dd,J=11.3,6.0Hz,1H),2.86(d,J=28.0Hz,2H),2.67 –2.64(m,2H),2.60(d,J=17.5Hz,1H),2.43(dd,J=13.1,4.4Hz,1H),2.33(d,J=23.7Hz,2H),2.01(dd,J=6. 4,4.1Hz,1H),1.90(d,J=10.5Hz,4H),1.69–1.61(m,4H),1.49(s,2H),1.34(d,J=6.7Hz,2H).ESI-MS[M+H] + m / z = 537.30.

[0278] Example 13: 3-(4-(5-(5-methyl-2H-spirocyclic[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0279]

[0280] The synthesis method and feed ratio were the same as in Example 1, yielding 32 mg of white solid, with a yield of 41%. 1 H NMR (500MHz, DMSO-d6) δ11.02(s,1H),7.95(s,1H),7.57(d,J=3.4Hz,1H),7.47–7.45(m,2H),7.16(s,1H),5. 16–5.12(m,1H),4.47(d,J=17.1Hz,1H),4.38(s,2H),4.31(d,J=17.1Hz,1H),2.96–2.90(m,1H),2.82(d,J=1 1.4Hz,2H),2.66(d,J=7.7Hz,2H),2.62–2.58(m,1H),2.46–2.41(m,1H),2.38(s,3H),2.33–2.29(m,2H),2.0 3–1.96(m,3H),1.82(t,J=10.9Hz,2H),1.65–1.60(m,4H),1.52–1.47(m,2H),1.35–1.31(m,2H).ESI-MS[M+H] + m / z = 537.30.

[0281] Example 14: 3-(1-oxo-4-(5-(5-(trifluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)isoindoline-2-yl)piperidin-2,6-dione

[0282]

[0283] The synthesis method and feed ratio were the same as in Example 1, yielding 35 mg of white solid, with a yield of 41%. 1 H NMR (500MHz, DMSO-d6) δ11.02(s,1H),8.29(s,1H),7.90(s,1H),7.58–7.55(m,1H),7.48–7.44(m,2H),5.14(dd,J=13.3,5. 1Hz,1H),4.57(s,2H),4.47(d,J=17.1Hz,1H),4.31(d,J=17.1Hz,1H),2.96–2.89(m,1H),2.83(d,J=6.0Hz,2H),2.67–2.63( m,2H),2.60(d,J=16.5Hz,1H),2.43(dd,J=13.1,4.4Hz,1H),2.28(t,J=7.3Hz,2H),2.03–1.98(m,1H),1.98–1.89(m,4H),1. 68(d,J=10.3Hz,2H),1.62(dd,J=15.2,7.7Hz,2H),1.48(dd,J=14.6,7.5Hz,2H),1.34(dd,J=14.8,7.6Hz,2H).ESI-MS[M+H] + m / z = 571.76.

[0284] Example 15: 3-(4-(5-(5-methoxy-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0285]

[0286] The synthesis method and feed ratio were the same as in Example 1, yielding 18 mg of white solid, with a yield of 23%. 1H NMR (500MHz, DMSO-d6) δ11.01(s,1H),7.61(s,1H),7.57(dd,J=5.1,3.5Hz,1H),7.47–7.45(m,2H),6.75(s,1H),5.14( dd,J=13.3,5.1Hz,1H),4.47(d,J=17.1Hz,1H),4.38(s,2H),4.31(d,J=17.1Hz,1H),3.75(s,3H),2.96–2.90(m,1H),2. 83(s,2H),2.66(d,J=7.6Hz,2H),2.60(d,J=17.2Hz,1H),2.43(dd,J=13.1,4.4Hz,1H),2.32(s,2H),2.06–1.92(m,3H), 1.85(t,J=10.9Hz,2H),1.63(dd,J=15.0,8.8Hz,4H),1.49(d,J=6.6Hz,2H),1.33(dd,J=14.5,7.5Hz,2H).ESI-MS[M+H] + m / z = 533.80.

[0287] Example 16: 3-(4-(5-(5-chloro-2H-spirocyclic [furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-6-fluoro-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0288]

[0289] The synthesis method and feed ratio were the same as in Example 1, yielding 28 mg of white solid, with a yield of 34%. 1H NMR(500MHz,DMSO-d6)δ11.03(s,1H),7.93(s,1H),7.47(s,1H),7.37–7.34(m,2H),5.14(dd,J=13.3,5.1Hz ,1H),4.49(s,2H),4.46(d,J=17.1Hz,1H),4.29(d,J=17.1Hz,1H),2.95–2.89(m,1H),2.82(d,J=9.2Hz,2H), 2.67–2.64(m,2H),2.60(d,J=16.5Hz,1H),2.42(dd,J=13.1,4.4Hz,1H),2.30(s,2H),2.04–2.00(m,1H),1.8 9(dd,J=26.8,15.2Hz,4H),1.65(dd,J=21.9,9.8Hz,4H),1.51–1.46(m,2H),1.35–1.31(m,2H).ESI-MS[M+H] + m / z = 555.37

[0290] Example 17: 3-(6-fluoro-1-oxo-4-(5-(5-(trifluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)isoindoline-2-yl)piperidin-2,6-dione

[0291]

[0292] The synthesis method and feed ratio were the same as in Example 1, yielding 36 mg of white solid, with a yield of 41%. 1 H NMR(500MHz,DMSO-d6)δ11.03(s,1H),8.28(s,1H),7.89(s,1H),7.37–7.34(m,2H),5.14(dd,J=13.3,5.1Hz, 1H),4.57(s,2H),4.46(d,J=17.1Hz,1H),4.30(d,J=17.1Hz,1H),2.96–2.88(m,1H),2.82(d,J=5.6Hz,2H),2. 68–2.64(m,2H),2.63–2.58(m,1H),2.42(dt,J=13.3,8.9Hz,1H),2.28(t,J=7.3Hz,2H),2.05–1.99(m,1H),1 .93(dd,J=10.0,3.4Hz,4H),1.70–1.61(m,4H),1.51–1.45(m,2H),1.34(dd,J=14.7,7.6Hz,2H).ESI-MS[M+H] + m / z = 589.27

[0293] Example 18: 3-(6-fluoro-4-(5-(5-fluoro-2H-spirocyclic [furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0294]

[0295] The synthesis method and feed ratio were the same as in Example 1, yielding 37 mg of white solid, with a yield of 46%. 1 H NMR (500MHz, DMSO-d6) δ11.02(s,1H),7.66(d,J=1.3Hz,1H),7.36(dd,J=6.6,4.3Hz,2H),7.17(s,1H),5.14(dd,J= 13.3,5.1Hz,1H),4.50–4.43(m,3H),4.29(d,J=17.1Hz,1H),2.96–2.90(m,1H),2.83(s,2H),2.68–2.64(m,2H),2.6 0(d,J=16.0Hz,1H),2.42(dd,J=13.1,4.4Hz,1H),2.31(s,2H),2.02(dd,J=9.0,3.6Hz,1H),1.89(t,J=11.0Hz,4H), 1.69(d,J=12.0Hz,2H),1.63(dd,J=15.1,7.6Hz,2H),1.53–1.46(m,2H),1.34(dd,J=14.7,7.6Hz,2H).ESI-MS[M+H] + m / z = 539.28.

[0296] Example 19: 3-(4-(5-(5-(difluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-6-fluoro-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0297]

[0298] The synthesis method and feed ratio were the same as in Example 1, yielding 41 mg of white solid, with a yield of 48%. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),8.21(s,1H),7.65(s,1H),7.40–7.33(m,2H),6.85(t,J=55.4Hz, 1H),5.14(dd,J=13.3,5.1Hz,1H),4.54(s,2H),4.50–4.26(m,2H),2.96–2.86(m,3H),2.66(t,J=7.7Hz ,2H),2.64–2.57(m,1H),2.47–2.35(m,3H),2.12–1.99(m,3H),1.93(ddd,J=14.9,12.0,3.5Hz,2H),1. 71(d,J=12.9Hz,2H),1.64(p,J=7.6Hz,2H),1.51(p,J=7.4Hz,2H),1.35(q,J=7.6Hz,2H).ESI-MS[M+H] + m / z = 571.61.

[0299] Example 20: (S)-3-(6-fluoro-1-oxo-4-(5-(5-(trifluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidine]-1)'-yl)pentyl)isoindoline-2-yl)piperidine-2,6-dione

[0300]

[0301] The synthesis method and feed ratio were the same as in Example 1, yielding 27 mg of white solid, with a yield of 31%. 1 H NMR(500MHz,DMSO-d6)δ11.01(s,1H),8.29(s,1H),7.87(s,1H),7.37–7.33(m,2H),5.14(dd,J=13.3,5.1Hz ,1H),4.58(s,2H),4.46(d,J=17.1Hz,1H),4.30(d,J=17.1Hz,1H),2.96–2.90(m,1H),2.89–2.82(m,2H),2.6 8–2.64(m,2H),2.60(dd,J=15.5,2.0Hz,1H),2.46–2.40(m,1H),2.39–2.28(m,2H),2.05–2.02(m,1H),2.01 –1.87(m,4H),1.70(d,J=10.0Hz,2H),1.66–1.61(m,2H),1.54–1.47(m,2H),1.37–1.31(m,2H).ESI-MS[M+H] + m / z = 589.27

[0302] Example 21: (S)-3-(4-(5-(5-chloro-2H-spirocyclic [furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0303]

[0304] The synthesis method and feed ratio were the same as in Example 1, yielding 25 mg of white solid, with a yield of 31%. 1 H NMR (500MHz, DMSO-d6) δ11.00(s,1H),7.92(s,1H),7.58–7.56(m,1H),7.46(s,2H),7.45(s,1H),5.14(dd,J=13.3,5 .1Hz,1H),4.47(d,J=17.2Hz,3H),4.31(d,J=17.1Hz,1H),2.96–2.89(m,1H),2.81(d,J=9.1Hz,2H),2.66(d,J=7.6Hz ,2H),2.62(dd,J=9.0,6.6Hz,1H),2.43(dd,J=13.1,4.4Hz,1H),2.29(s,2H),2.04–2.00(m,1H),1.89(dd,J=26.5,1 4.6Hz,4H),1.66(d,J=11.6Hz,2H),1.64–1.60(m,2H),1.51–1.46(m,2H),1.34(dd,J=14.8,7.7Hz,2H).ESI-MS[M+H] + m / z = 537.29

[0305] Example 22: (S)-3-(4-(5-(5-chloro-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-6-fluoro-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0306]

[0307] The synthesis method and feed ratio were the same as in Example 1, yielding 31 mg of white solid, with a yield of 37%. 1H NMR (500MHz, DMSO-d6) δ11.01(s,1H),7.92(s,1H),7.46(s,1H),7.35(dd,J=6.4,4.2Hz,2H),5.14(dd,J=1 3.3,5.1Hz,1H),4.50–4.43(m,3H),4.30(d,J=17.1Hz,1H),2.96–2.89(m,1H),2.82(d,J=8.5Hz,2H),2.68 –2.63(m,2H),2.61(d,J=16.4Hz,1H),2.45–2.38(m,1H),2.29(s,2H),2.05–2.01(m,1H),1.89(dd,J=26.3 ,14.4Hz,4H),1.65(dd,J=19.4,10.7Hz,4H),1.52–1.45(m,2H),1.33(dd,J=14.7,7.6Hz,2H).ESI-MS[M+H] + m / z = 555.37.

[0308] Example 23: (S)-3-(1-oxo-4-(5-(5-(trifluoromethyl)-2H-spiro[furan[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)isoindoline-2-yl)piperidin-2,6-dione

[0309]

[0310] The synthesis method and feed ratio were the same as in Example 1, yielding 29 mg of white solid, with a yield of 34%. 1 H NMR (500MHz, DMSO-d6) δ11.00(s,1H),8.28(s,1H),7.89(s,1H),7.58–7.56(m,1H),7.47–7.45(m,2H),5.14(dd,J= 13.3,5.1Hz,1H),4.57(s,2H),4.47(d,J=17.1Hz,1H),4.31(d,J=17.1Hz,1H),2.96–2.89(m,1H),2.83(s,2H),2.67 –2.63(m,2H),2.60(dd,J=15.6,2.0Hz,1H),2.43(dd,J=13.1,4.5Hz,1H),2.29(s,2H),2.04–2.00(m,1H),1.94(d,J =8.6Hz,4H),1.68(d,J=9.6Hz,2H),1.65–1.60(m,2H),1.51–1.46(m,2H),1.34(dd,J=9.8,4.8Hz,2H).ESI-MS[M+H]+ m / z = 571.76.

[0311] Example 24: (S)-1'-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)pentyl)-2H-spiro[furan[2,3-c]pyridine-3,4'-piperidin]-5-carboxynitrile

[0312]

[0313] The synthesis method and feed ratio were the same as in Example 1, yielding 10 mg of white solid, with a yield of 13%. 1 H NMR (500MHz, DMSO-d6) δ11.00(s,1H),8.31(s,1H),8.07(s,1H),7.59–7.55(m,1H),7.46(d,J=4.0Hz,2H),5.14(dd ,J=13.3,5.2Hz,1H),4.58(s,2H),4.47(d,J=17.1Hz,1H),4.31(d,J=17.1Hz,1H),2.97–2.89(m,1H),2.82(d,J=9.3 Hz,2H),2.70–2.63(m,2H),2.63–2.58(m,1H),2.43(dd,J=13.1,4.5Hz,1H),2.32–2.23(m,2H),2.02(ddd,J=10.4,5 .2,3.0Hz,1H),1.99–1.84(m,4H),1.70–1.59(m,4H),1.52–1.45(m,2H),1.34(dd,J=14.8,7.7Hz,2H).ESI-MS[M+H] + m / z = 528.36.

[0314] Example 25: (S)-1'-(5-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1-oxoisodihydroindole-4-yl)pentyl)-2H-spiro[furan[2,3-]c]pyridine-3,4'-piperidin]-5-carboxynitrile

[0315]

[0316] The synthesis method and feed ratio were the same as in Example 1, yielding 16 mg of white solid, with a yield of 20%. 1H NMR (500MHz, DMSO-d6) δ11.01(s,1H),8.31(s,1H),8.07(s,1H),7.35(dd,J=6.5,4.3Hz,2H),5.14(dd,J=13.3,5 .1Hz,1H),4.58(s,2H),4.46(d,J=17.1Hz,1H),4.30(d,J=17.1Hz,1H),2.96–2.88(m,1H),2.82(d,J=10.5Hz,2H) ,2.68–2.63(m,2H),2.61(d,J=17.5Hz,1H),2.42(dt,J=13.4,8.9Hz,1H),2.29(t,J=7.3Hz,2H),2.02(dd,J=8.8 ,3.7Hz,1H),2.00–1.85(m,4H),1.70–1.61(m,4H),1.53–1.44(m,2H),1.34(dd,J=14.4,7.6Hz,2H).ESI-MS[M+H] + m / z = 546.33.

[0317] The compounds in the following examples can be obtained by following synthesis method one and synthesis method two.

[0318] Example 26: 3-(4-(5-(7-chloro-5-fluoro-2H-spiro[furano[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0319]

[0320] Example 27: 3-(4-(5-(5,7-difluoro-2H-spiro[furano[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0321]

[0322] Example 28: 3-(4-(5-(7-chloro-5-fluoro-2H-spiro[furano[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-6-fluoro-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0323]

[0324] Example 29: 3-(4-(5-(5,7-difluoro-2H-spiro[furano[2,3-c]pyridin-3,4'-piperidin]-1'-yl)pentyl)-6-fluoro-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0325]

[0326] Example 30: 3-(4-(5-(5-methyl-2H-spiro[furano[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0327]

[0328] Example 31: 3-(6-fluoro-4-(5-(5-methyl-2H-spiro[furano[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0329]

[0330] Example 32: 3-(4-(5-(5-fluoro-2H-spiro[furano[2,3-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0331]

[0332] Example 33: 3-(6-fluoro-4-(5-(5-fluoro-2H-spiro[furano[2,3-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione

[0333]

[0334] Example 34: 3-(1-oxo-4-(5-(6-(trifluoromethyl)-2H-spiro[furano[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)isoindoline-2-yl)piperidin-2,6-dione

[0335]

[0336] Example 35: 3-(6-fluoro-1-oxo-4-(5-(6-(trifluoromethyl)-2H-spiro[furano[3,2-b]pyridin-3,4'-piperidin]-1'-yl)pentyl)isoindoline-2-yl)piperidin-2,6-dione

[0337]

[0338] II. Experimental Examples

[0339] This invention also tested the activity of the multisubstituted isoindoline compounds of the embodiments against three major hematologic malignancy cell lines: acute myeloid leukemia cell line (MV-4-11), multiple myeloma cell line (MM.1S), and mantle cell lymphoma cell line (Mino). The cell proliferation inhibitory activity of these three representative cell lines was tested. Unless otherwise specified, all experimental materials required for the pharmacological experiments were commercially available.

[0340] 1. Inhibitory activity of the compound on the proliferation of MV-4-11 cells

[0341] MV-4-11 cells were cultured and collected using IMDM supplemented with 10% fetal bovine serum. 180 μL of cell suspension was added to each well of a 96-well cell culture plate to achieve a cell count of 2000. 20 μL of DMSO (final concentration 0.2%) was added to the control cell wells. The compound was serially diluted 5-fold from a 10 mM stock solution, with 20 μL added to each well (DMSO final concentration 0.2%). Cells were incubated at 37°C in a 5% CO2 incubator for 7 days. The reaction solution was prepared according to the MTS kit (Promega, G5430), and 20 μL was added to each well. Cells were incubated at 37°C in a 5% CO2 incubator for 3–4 hours. The absorbance at 490 nm was read using a microplate, with the 690 nm absorbance as the background value. OD490–OD690 was used as the final raw data. The inhibition rate of the compound was calculated using the formula: Inhibition rate = (OD690–OD490) / (OD690–OD490) = (OD690–OD490) / (OD690–OD490). DMSO -OD 化合物 ) / (OD DMSO -O 空白 ()×100%. The proliferation inhibition IC50 of the compound. 50 The results were fitted using Graph Pad Prism 5.0. The experiment was repeated three times, with three parallel experiments used for each iteration to calculate the mean and standard deviation.

[0342] The results of the MV-4-11 cell viability assay are shown in Table 1 below: A represents the cell viability IC50. 50 <60nM, B indicates cell viability 60nM≤IC 50 ≤200 nM, C indicates cell viability 200 nM <IC 50 ≤10μM; D indicates cell viability >10μM.

[0343] Table 1. Inhibitory activity of compounds on MV-411 cell proliferation

[0344]

[0345]

[0346] Based on the cell growth inhibitory activity test results of the above compounds, the compounds of the embodiments of the present invention exhibit excellent inhibitory activity against the cell growth of the acute myeloid leukemia cell line (MV-4-11), with an inhibitory activity level of A or B. More importantly, the activity of the compounds provided by the present invention is significantly superior to the marketed drugs lenalidomide or pomalidomide, both of which have an inhibitory activity level of D in MV-4-11 cells. The activity test results of the compounds of the present invention against multiple myeloma (MM.1S) and mantle cell lymphoma (Mino) cells also show that the compounds have good activity.

[0347] 2. Activity of the compound on peripheral blood mononuclear cells (PBMCs)

[0348] PBMC cells were cultured and collected using RPMI-1640 with 10% fetal bovine serum. 90 μL of cell suspension was added to each well of a 96-well cell plate to achieve a cell count of 10,000. 10 μL of DMSO (final concentration 0.2%) was added to the control cell wells. The compound was serially diluted 5-fold from a 10 mM stock solution, with 10 μL added to each compound cell well (DMSO final concentration 0.2%). Cells were incubated at 37°C in a 5% CO2 incubator for 3 days. 30 μL of the reaction solution was added to each well according to the CTG kit (Promega, G7572), and the cells were incubated at room temperature on a shaker at low speed for 10 min. The luminescence value was read using Envision. The inhibition rate of the compound was calculated as: Inhibition rate = (DMSO - Compound) / (DMSO - Blank) × 100%. The IC50 of the compound's proliferation inhibition was... 50 The results were fitted using Graph Pad Prism 8.0. The experiment was repeated three times, with three parallel experiments used for each iteration to calculate the mean and standard deviation.

[0349] 3. Experimental methods and results regarding the effects of the compounds of this invention on hERG potassium channels.

[0350] Experimental platform: Qpatch automated patch clamp system

[0351] Cell line: Chinese hamster ovary (CHO) cells stably expressing hERG potassium channels

[0352] Experimental methods: CHO-hERG cells were cultured at 175 cm⁻¹. 2In the culture flask, once the cell density reaches 60-80%, remove the culture medium, wash once with 7 mL of PBS (Phosphate Buffered Saline), and then add 3 mL of L-Detachin for digestion. After complete digestion, add 7 mL of culture medium to neutralize, then centrifuge, aspirate the supernatant, and resuspend in 5 mL of culture medium to ensure a cell density of 2-5 × 10⁶ cells / year. 6 / mL. Single-cell high-impedance sealing and whole-cell pattern formation were automated using the Qpatch instrument. After obtaining the whole-cell recording pattern, cells were clamped at -80 mV. Before a 5-second +40 mV depolarization stimulus, a 50-millisecond -50 mV pre-voltage was applied, followed by repolarization to -50 mV for 5 seconds, then back to -80 mV. This voltage stimulus was applied every 15 seconds. After 2 minutes of recording, extracellular fluid was administered for 5 minutes, followed by drug delivery. The compound concentration was started from the lowest test concentration, and each test concentration was administered for 2.5 minutes. After all concentrations were administered, the positive control compound Cisapride was given. At least 3 cells (n>=3) were tested for each compound concentration. The extracellular fluid formulation (mM) consisted of 140 NaCl, 5 KCl, 1 CaCl2, 1.25 MgCl2, 10 HEPES, and 10 Glucose, adjusted to pH 7.4 with NaOH. Intracellular fluid formulation (mM): 140 KCl, 1 MgCl2, 1 CaCl2, 10 EGTA and 10 HEPES, pH adjusted to 7.2 with KOH.

[0353] Data processing:

[0354] Data analysis and processing were performed using Assay Software v5.6.4, GraphPad Prism 8, and Excel. Different compounds...

[0355] The degree to which the concentration of the substance inhibits the hERG potassium current (the peak value of the hERG tail current induced at -50mV) is measured by the following...

[0356] Formula calculation: Inhibition% = [1 – (I / Io)] × 100%

[0357] Where Inhibition% represents the percentage of inhibition of the hERG potassium current by the compound, and I and Io are respectively

[0358] This indicates the amplitude of the hERG potassium current before and after drug administration.

[0359] Compound IC 50 The following equations were fitted and calculated using GraphPad Prism 8 software:

[0360] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0361] Where X is the Log value of the detected concentration of the test sample, Y is the inhibition percentage at the corresponding concentration, and Bottom

[0362] Top and Minimum and Maximum Suppression Percentages, respectively.

[0363] 4. Experimental Results:

[0364] IC of the inhibitory effect of the compound of the present invention on hERG potassium channel current 50 The values ​​are shown in Table 2 below: A represents IC. 50 ≥10μM, B indicates 4μM <IC 50 <10μM, C means 1.5μM≤IC 50 ≤4μM, D indicates IC 50 <1.5μM.

[0365] Table 2: Results of the inhibitory activities of the compounds on PMBC cells and hERG cells

[0366]

[0367]

[0368] Based on the above results of the activity test of the compounds on PBMC cells, the compounds of the present invention have an activity greater than 20 micromoles (μM) on PBMC cells, indicating that the compounds of the present invention are not toxic to PBMC cells.

[0369] In recent years, some drugs have been withdrawn from the market due to their inhibitory effects on hERG channels. Therefore, reducing the inhibitory activity of drugs against hERG is an important issue to consider in new drug development. Most of the compounds in the embodiments of this invention exhibit hERG inhibitory activity greater than 4 micromoles (μM), IC50. 50 A rating of A or B indicates weak inhibitory activity against hERG. In contrast, the comparative compounds 123, 127, 129, and 157 in the prior art (CN114085212B) all have an IC50 rating of D. 50 <1.5 μM). As can be seen from Tables 1 and 2 above, the compounds of the embodiments of the present invention significantly reduce the inhibitory activity against hERG without reducing tumor suppressor activity, which can significantly reduce the risk of cardiotoxicity caused by the compounds and greatly improve the safety of the compounds in subsequent clinical use.

[0370] 5. In vitro cell degradation data of compounds

[0371] Western blot analysis

[0372] MV-4-11 cells in logarithmic growth phase were harvested at a concentration of 5 × 10⁻⁶ cells / cells. 5 Cells were seeded per well in 12-well plates. Different concentrations of compounds diluted with complete culture medium were added to a final volume of 1 mL. After culturing for 16 h, cells were collected into 2 mL EP tubes, centrifuged to remove the supernatant, and then lysed on ice for 30 min with 100 μL of 1× Loading buffer. The cells were then boiled in a metal bath at 95 °C for 15 min. After cooling, the samples were subjected to SDS-PAGE electrophoresis and transferred to nitrocellulose membranes. The membranes were blocked with 5% skim milk powder at room temperature for 1.5 h. Primary antibodies (GSPT1, IKZF1, CK1α, and β-actin) were added and the membranes were incubated overnight at 4 °C. After washing 10 min × 3 times, the corresponding secondary antibodies (1:10000) were added and the membranes were incubated at room temperature for 1 h. After washing 10 min × 3 times, ECL developing solution was added, and the membranes were exposed using a gel imaging system. The bands were processed and analyzed using ImageLab software. The gray value of the band represents the signal intensity; the lower the gray value, the lower the signal intensity. The expression level (Int) of each protein was calculated as the relative fold increase normalized to the internal control protein. The degradation rate of the target protein in each group was calculated as the ratio of the target protein expression level (Int_target) in the compound group to the target protein expression level (Int_DMSO) in the negative control group. The calculation formula is as follows: Protein degradation = (1 - Int_target / Int_DMSO) × 100%

[0373] The degradation activities of IKZF1 and CK1α are shown in Table 3. In Table 3, A represents DC. 50 <10nM, B means 10nM ≤ DC 50 ≤100nM, where C represents 100nM < DC 50 <1000nM, D represents DC 50 >1000nM; D max "a" means >80%; "b" means from >60% to <80%; "c" means <60%.

[0374] The degradation activity of GSPT1 is shown in Table 4. In Table 4, the % degradation values ​​are reported as "A", "B", "C" or "D". "A" indicates that the % degradation value is greater than 75% (value > 75%); "B" indicates that the % degradation value is equal to or greater than 50% and less than 75% (50% ≤ value < 75%); "C" indicates that the % degradation value is equal to or greater than 25% and less than 50% (25% ≤ value < 50%); "D" indicates that the % degradation value is less than 25% (< 25%).

[0375] Table 3: Degradation results of the compounds on CK1α and IKZF1 in MV-4-11 cells

[0376]

[0377] Based on the degradation results of the above compounds on CK1α and IKZF1 in MV-4-11 cells, it is shown that the compounds in the embodiments of the present invention can efficiently degrade CK1α (DC). 50 <10nM,D max The CK1α degradation efficiency is >80%, while the IKZF1 degradation efficiency is relatively weak. Therefore, the compounds of this invention, through the introduction of a pyridine ring into their molecular structure, have an unexpected advantage: they can effectively improve the selectivity for CK1α degradation while reducing the degradation of other proteins such as IKZF1. Thus, the compounds provided by this invention can primarily degrade CK1α, reducing the side effects of degrading other proteins, such as neutropenia.

[0378] Table 4: Degradation of GSPT1 protein in MV-4-11 cells by the compounds

[0379]

[0380]

[0381] Based on the degradation results of the above compounds on GSPT1 in MV-4-11 cells, it is shown that the compounds of the present invention do not degrade GSPT1 at 1 μM. Therefore, the pyridine ring-substituted isoindoline compounds provided by the present invention can reduce the toxic side effects on normal cells caused by the degradation of GSPT1.

[0382] In summary, this invention provides a new class of compounds with excellent antitumor activity but significantly reduced toxicity. Compared with the superior compounds 123, 127, 129, and 157 in the prior art (CN114085212B), the compounds of this invention exhibit comparable or superior inhibitory activity against the acute myeloid leukemia cell line (MV-4-11), and are significantly superior to the marketed drugs lenalidomide or pomalidomide. Therefore, the compounds of this invention can expand the indications for hematologic malignancies where existing thalidomide drugs are ineffective, such as their application in the treatment of acute myeloid leukemia.

[0383] Even more surprisingly, the introduction of a pyridine ring into the molecular structure resulted in a significant reduction in hERG-induced cardiotoxicity in the pyridine heterocyclic-substituted isoindoline compounds of this invention. This was a highly unexpected and outstanding effect. Those skilled in the art will recognize that drug-induced cardiotoxicity is a significant concern in recent years, a major reason for drug withdrawal from the market. The hERG potassium channel is an important target for drug-induced arrhythmias and antiarrhythmic drugs. Therefore, reducing the inhibitory activity of drugs against hERG is a crucial issue to consider in new drug development. The majority of the compounds in the embodiments of this invention exhibit an IC50 value for hERG. 50 The concentration greater than 4 micromolar (μM) exhibits weak inhibitory activity against hERG, while the IC50 of non-pyridine-substituted comparative compounds 123, 127, 129, and 157 in the prior art (CN114085212B) is significantly higher. 50 With a concentration generally <1.5 μM, it is evident that the compounds of this invention exhibit significantly lower inhibitory activity against hERG, thereby reducing the risk of potential cardiotoxicity and greatly improving the safety of the compounds in subsequent clinical use.

[0384] Furthermore, the compounds of this invention can efficiently degrade CK1α with good selectivity, while reducing degradation of other proteins such as IKZF1 and GSPT1. Therefore, another unexpected and significant advantage of the compounds provided by this invention is their selective degradation of CK1α, thereby reducing the side effects of degrading other proteins, such as neutropenia and reducing the toxic side effects on normal cells caused by GSPT1 degradation.

[0385] In summary, the objective of this invention is to provide a novel CRL4 structure. CRBN E3 ubiquitin ligase molecular gel degraders can improve the therapeutic effect of tumors, reduce toxic side effects, and broaden the indications for new molecular gel degraders based on CRL4CRBNE3 ubiquitin ligases.

[0386] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of formula (I), including its pharmaceutically acceptable salt, tautomer, or stereoisomer: in, X is CHR3, CR3R4, or CO; Each of R3 and R4 is independently hydrogen or a C1-C6 alkyl group; R1 is hydrogen or halogen; R2 is hydrogen, fluorine, or a C1-C6 alkyl group; Z can be (CH2)n, -O-(CH2)n, or -NH-(CH2)n; n can be 4, 5, or 6 independently; Y is hydrogen, halogen, C1-C6 alkyl, or C1-C6 haloalkyl; Y1 is 0; X1 is either C-R5 or N; X2 is either C-R6 or N; X3 is either C-R7 or N; X4 is either C-R8 or N; Among X1, X2, X3, and X4, only one is N; R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, nitro, hydroxyl, carboxyl, aminocarbonyl, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

2. The compound according to claim 1, characterized in that, The compound has the structure shown in formula (I-1): Where X represents CHR3, CR3R4, and CO; Each of R3 and R4 is hydrogen; R1 is hydrogen or fluorine; R2 is hydrogen, fluorine, or methyl; Z can be (CH2)n, -O-(CH2)n, or -NH-(CH2)n; n is 4, 5, or 6; Y represents hydrogen; X1 is either C-R5 or N; X2 is either C-R6 or N; X3 is either C-R7 or N; X4 is either C-R8 or N; Among X1, X2, X3, and X4, only one is N; R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, hydroxyl, aminocarbonyl, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

3. The compound according to claim 1, characterized in that, The compound has the structure shown in formula (I-2): Where R1 is hydrogen or fluorine; Z is (CH2)n or -O-(CH2)n; n is 4, 5, or 6; Y represents hydrogen; X1 is either C-R5 or N; X2 is either C-R6 or N; X3 is either C-R7 or N; X4 is either C-R8 or N; Among X1, X2, X3, and X4, only one is N; R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

4. The compound according to claim 1, characterized in that, The compound has the structure shown in formula (I-3): R1 is hydrogen or fluorine; n is 4, 5, or 6; Y represents hydrogen; X1 is either C-R5 or N; X2 is either C-R6 or N; X3 is either C-R7 or N; X4 is either C-R8 or N; Among X1, X2, X3, and X4, only one is N; R5, R6, R7, and R8 are each independently selected from: hydrogen, halogen, cyano, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy, and halogen-substituted C1-C6 alkoxy.

5. The compound according to any one of claims 1-4, characterized in that, Selected from:

6. The compound according to claim 1, characterized in that, The compounds are selected from the group consisting of:

7. A method for preparing the compound of claim 1, wherein, The method is selected from one of the following methods: X, X1, X2, X3, X4, Y, R1, and n are the same as those defined in claim 3, wherein X5 is CH2 or O; Step 1-1: Compounds 1A and 1B react with potassium carbonate in DMF solution by heating to obtain compound 1C; Steps 1-2: Compound 1C, tri-n-butyltin hydrogen and azobisisobutyronitrile were reacted in toluene at 135±10℃ to give compound 1D; Steps 1-3: Compound 1D reacts with dioxane hydrochloride at room temperature to give compound 1F; Steps 1-4: Compounds 1G and 1F reacted with triethylamine at room temperature to give compound 1H; Synthesis Method Two: X1, X2, X3, X4, n, and Y are the same as those defined in claim 4; Step 2-1: Compound 2A and compound 2B react in the presence of N,N-diisopropylethylamine to give compound 2C; Step 2-2: Compound 2C reacts with carbon tetrabromide and triphenylphosphine to give compound 2D; Steps 2-3: Compounds 2D and 1F reacted with triethylamine at room temperature to give compound 2E.

8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the compound of any one of claims 1-6, a pharmaceutically acceptable salt, tautomer or stereoisomer, and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to claim 8, characterized in that, The pharmaceutical composition further comprises one or more selected from dexamethasone, rituximab, trastuzumab, PD-1 inhibitors, PD-L1 inhibitors, pemetrexed, topotecan, doxorubicin, gemcitabine, dacarbazine, clarithromycin, vincristine, cytarabine, azacitidine, prednisone, docetaxel, clofarabine injection, HDAC inhibitors, androgen receptor inhibitors, androgen biosynthesis inhibitors, BTK inhibitors, BCL2 inhibitors, erythrocyte growth hormone, minocycline, elotuzumab, palbociclib, nivolumab, pembrolizumab, panobinostat, ubliximab, romidepsin, eltrombopag, CAR-T, and melphalan.

10. The compound of any one of claims 1-6, wherein a pharmaceutically acceptable salt, tautomer, or stereoisomer, or pharmaceutical combination of any one of claims 8-9, is used in the preparation for the prevention or treatment of CRL4. CRBN Use in medicines for diseases related to E3 ubiquitin ligases, preferably, those related to CRL4 CRBN Diseases associated with E3 ubiquitin ligase include cancer, pain, inflammation, central nervous system disorders, or immune system disorders.

11. The use as described in claim 10, characterized in that, The cancers mentioned are hematologic malignancies and solid tumors. Preferably, the hematologic malignancies are selected from: myelodysplastic syndromes, multiple myeloma, mantle cell lymphoma, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, myelofibrosis, Burkitt lymphoma, Hodgkin lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, ciliary body and chronic melanoma, iris melanoma, relapsed interocular melanoma, T-cell lymphoma, erythroid lymphoma, monocytic and monocyte leukemia, myeloid leukemia, and central nervous system lymphoma. The solid tumors are selected from: meningeal malignancies. Tumors, spinal cord tumors, lung cancer, ovarian cancer, skin cancer, renal cell carcinoma, astrocytoma, amyloidosis, type I complex regional pain syndrome, malignant melanoma, radiculopathy, glioblastoma, sarcoma, malignant glioma, refractory plasmacytoma, extraocular melanoma, thyroid cancer, breast cancer, prostate cancer, hepatocellular carcinoma or primary macroglobulinemia, adrenocortical carcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colorectal cancer, esophageal cancer, gallbladder cancer, gastric cancer, head and neck cancer, kidney cancer, liver cancer, uterine cancer, rhabdomyosarcoma, neuroblastoma, pancreatic cancer, testicular cancer; The pain is nociceptive pain, neuropathic pain, visceral pain, migraine, headache, or postoperative pain; The neuropathic pain mentioned is complex regional pain syndrome, malnutrition, or diabetic neuropathy; The central nervous system diseases mentioned are amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis, or Tourette syndrome.

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