A GSPT1 degradation agent containing an alkyne group and its application

Abstract

The present invention relates to a compound of formula I or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative thereof. The compound has the activity of degrading GSPT1 protein and can be used to prevent and treat GSPT1 protein-related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and particularly relates to an alkyne-containing GSPT1 degradation agent and application thereof. Background Art

[0002] The ubiquitin-proteasome system (UPS) is the primary pathway for protein degradation in most cells and plays a crucial role in numerous cellular activities, including DNA repair, signal transduction, transcription and translation, and immune responses. By hijacking the inherent UPS in organisms, pathogenic proteins can be rapidly destroyed and eliminated, thereby achieving targeted protein degradation (TPD). In recent years, the most widely studied and mature TPD technologies fall into two categories: proteolysis-targeting chimeras (PROTACs) and molecular glues. Compared to PROTACs, molecular glues have lower molecular weights and often possess higher membrane permeability and favorable pharmacokinetic properties, making them promising small-molecule protein degraders.

[0003] Molecular glues bind to E3 ligases and alter their surface and specificity, leading to the recruitment, ubiquitination, and degradation of substrates not normally targeted by the ligase (neosubstrates). Recognition of new substrates is controlled by protein-ligase surface interactions and does not require a ligand binding pocket. Therefore, molecular glues can be used to degrade undruggable targets, such as the translation termination factor G1 to S phase transition 1 (GSPT1).

[0004] GSPT1 mediates stop codon recognition by binding to eukaryotic release factor 1 (eRF1), promoting the release of nascent peptides from the ribosome and thus terminating protein translation. It is also involved in several other key cellular processes, such as cell cycle regulation, cytoskeleton formation, and apoptosis. GSPT1 expression is closely associated with tumor development and progression, and is considered an oncogenic driver in various cancers, such as breast, liver, gastric, and prostate cancers. Studies have shown that GSPT1 degradation leads to impaired translation termination, activation of the integrated stress response pathway, and TP53-independent cell death. Studies have also shown that Myc-driven tumors are critically dependent on the transcription factor Myc for protein synthesis, and GSPT1 is a terminator that regulates the translation process, making Myc-driven tumors more sensitive to GSPT1-targeted therapies.

[0005] GSPT1 is not only closely associated with tumor development and progression but also plays a crucial role in the viral lifecycle. Viral replication is highly dependent on host factors, and GSPT1 is a key host protein regulating viral RNA synthesis. Lassa virus (LASV) and Ebola virus (EBOV), as RNA viruses, have life cycles that are highly dependent on the replication and transcriptional activity of virally encoded RNA-dependent RNA polymerases in the host cytoplasm. Studies have shown that GSPT1 physically interacts with LASV polymerase, acting as a proviral factor that promotes LASV replication. Proximity interactome analysis has also confirmed GSPT1 as a druggable target for host-targeted antiviral drugs. Studies have also shown that EBOV can promote viral replication by disrupting the cellular mRNA decay machinery and repurposing host factors such as GSPT1 and UPF1.

[0006] Currently, there are no selective GSPT1 degraders marketed worldwide. Only three small molecule compounds have entered early clinical trials: CC-90009 (Phase I / II, NCT04336982, parenteral administration, terminated), MRT-2359 (Phase I / II, NCT05546268, oral administration), and FD-001 (Phase I, CTR20233435, oral administration). In vitro, MRT-2359 demonstrated potent antiproliferative activity against Myc-overexpressing tumor cell lines and dose-dependent tumor inhibition in mouse tumor models. Furthermore, CC-90009 demonstrated significant inhibitory effects on both LASV and EBOV infection in cell culture models. In EBOV-infected human hepatocytes, the drug effectively reduced viral RNA load and viral particle production, further confirming the importance of GSPT1 in viral replication. The strategy of targeted degradation of GSPT1 provides a new direction for the development of anti-tumor and antiviral drugs. Its mechanism of action can avoid the limitations of traditional anti-tumor and antiviral drugs that are prone to induce drug-resistant mutations, and has important development value.

[0007] Given the therapeutic potential of GSPT1 molecular glue degraders in tumor and virus treatment and their early development stage, GSPT1 small molecule degraders still need in-depth research. Summary of the Invention

[0008] Problems to be solved by the invention

[0009] In order to solve the above problems, the present invention provides a class of degradation agents targeting GSPT1 protein, which can be used to prevent and treat GSPT1-related diseases.

[0010] Solutions for solving problems

[0011] The present invention provides a compound of formula I or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative thereof.

[0012]

[0013] in,

[0014] R is selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-6 membered heteroaryl and substituted or unsubstituted 4-6 membered heterocyclic group, wherein the heteroaryl and heterocyclic group have 1-4 heteroatoms selected from N, O, and S, and the substitution is substituted by at least one Q;

[0015] Each occurrence of Q is independently selected from hydrogen, deuterium, halogen, cyano, hydroxy, NR'(R"), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocyclic group and C 6-10 Aryl, 5-10 membered heteroaryl;

[0016] R', R'' are each independently selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl and C 1-3 Halogenated alkoxy.

[0017] Effects of the Invention

[0018] The compounds provided by the present invention have strong degradation activity against GSPT1 protein. For example, the degradation rate of compound 3 against GSPT1 at a concentration of 10 nM is 61.7%. At the same time, they have excellent efficacy, in vitro / in vivo pharmacokinetic properties and safety, and have high prospects for clinical application. DETAILED DESCRIPTION

[0019] To make the technical solutions and beneficial effects of the present invention more clearly understood, the following is a detailed description by listing specific embodiments. Unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application belongs.

[0020] The present invention provides a compound of formula I or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative thereof.

[0021]

[0022] in,

[0023] R is selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-6 membered heteroaryl and substituted or unsubstituted 4-6 membered heterocyclic group, wherein the heteroaryl and heterocyclic group have 1-4 heteroatoms selected from N, O, and S, and the substitution is substituted by at least one Q;

[0024] Each occurrence of Q is independently selected from hydrogen, deuterium, halogen, cyano, hydroxy, NR'(R"), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocyclic group and C 6-10 Aryl, 5-10 membered heteroaryl;

[0025] R', R'' are each independently selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl and C 1-3 Halogenated alkoxy.

[0026] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted C 1~5 Alkyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted 5- to 6-membered heteroaryl and substituted or unsubstituted 4- to 6-membered heterocyclic group, wherein the heteroaryl group has 1 to 2 heteroatoms selected from N, O, and S, and the heterocyclic group has 1 to 2 heteroatoms selected from N, O, and S.

[0027] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isopentyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperazinyl, and substituted or unsubstituted thiomorpholinyl.

[0028] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, and substituted or unsubstituted piperidinyl.

[0029] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), C 1~6 Alkyl and C 1~6 Alkoxy.

[0030] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), and C 1~3 alkyl.

[0031] In certain embodiments, each occurrence of Q is independently selected from hydroxy, NR'(R"), and methyl.

[0032] In certain embodiments, R' and R'' are each independently selected from hydrogen and C 1-3 alkyl.

[0033] In certain embodiments, R' and R" are each independently selected from hydrogen and methyl.

[0034] The present invention provides a compound of formula I-1 or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative thereof.

[0035]

[0036] in,

[0037] R is selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-6 membered heteroaryl and substituted or unsubstituted 4-6 membered heterocyclic group, wherein the heteroaryl and heterocyclic group have 1-4 heteroatoms selected from N, O, and S, and the substitution is substituted by at least one Q;

[0038] Each occurrence of Q is independently selected from hydrogen, deuterium, halogen, cyano, hydroxy, NR'(R"), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocyclic group and C 6-10 Aryl, 5-10 membered heteroaryl;

[0039] R', R'' are each independently selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl and C 1-3 Halogenated alkoxy.

[0040] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted C 1~5 Alkyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted 5- to 6-membered heteroaryl and substituted or unsubstituted 4- to 6-membered heterocyclic group, wherein the heteroaryl group has 1 to 2 heteroatoms selected from N, O, and S, and the heterocyclic group has 1 to 2 heteroatoms selected from N, O, and S.

[0041] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isopentyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperazinyl, and substituted or unsubstituted thiomorpholinyl.

[0042] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, and substituted or unsubstituted piperidinyl.

[0043] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), C 1~6 Alkyl and C 1~6 Alkoxy.

[0044] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), and C 1~3 alkyl.

[0045] In certain embodiments, each occurrence of Q is independently selected from hydroxy, NR'(R"), and methyl.

[0046] In certain embodiments, R' and R'' are each independently selected from hydrogen and C 1-3 alkyl.

[0047] In certain embodiments, R' and R" are each independently selected from hydrogen and methyl.

[0048] The present invention provides a compound of formula I-2 or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative thereof.

[0049]

[0050] in,

[0051] R is selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-6 membered heteroaryl and substituted or unsubstituted 4-6 membered heterocyclic group, wherein the heteroaryl and heterocyclic group have 1-4 heteroatoms selected from N, O, and S, and the substitution is substituted by at least one Q;

[0052] Each occurrence of Q is independently selected from hydrogen, deuterium, halogen, cyano, hydroxy, NR'(R"), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocyclic group and C 6-10 Aryl, 5-10 membered heteroaryl;

[0053] R', R'' are each independently selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl and C 1-3 Halogenated alkoxy.

[0054] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted C 1~5 Alkyl, substituted or unsubstituted C 3~6Cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted 5- to 6-membered heteroaryl and substituted or unsubstituted 4- to 6-membered heterocyclic group, wherein the heteroaryl group has 1 to 2 heteroatoms selected from N, O, and S, and the heterocyclic group has 1 to 2 heteroatoms selected from N, O, and S.

[0055] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isopentyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperazinyl, and substituted or unsubstituted thiomorpholinyl.

[0056] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, and substituted or unsubstituted piperidinyl.

[0057] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), C 1~6 Alkyl and C 1~6 Alkoxy.

[0058] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), and C 1~3 alkyl.

[0059] In certain embodiments, each occurrence of Q is independently selected from hydroxy, NR'(R"), and methyl.

[0060] In certain embodiments, R' and R'' are each independently selected from hydrogen and C 1-3 alkyl.

[0061] In certain embodiments, R' and R" are each independently selected from hydrogen and methyl.

[0062] The present invention provides a compound of formula I-3 or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative thereof.

[0063]

[0064] in,

[0065] R is selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-6 membered heteroaryl and substituted or unsubstituted 4-6 membered heterocyclic group, wherein the heteroaryl and heterocyclic group have 1-4 heteroatoms selected from N, O, and S, and the substitution is substituted by at least one Q;

[0066] Each occurrence of Q is independently selected from hydrogen, deuterium, halogen, cyano, hydroxy, NR'(R"), C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl, 3-12 membered heterocyclic group and C 6-10 Aryl, 5-10 membered heteroaryl;

[0067] R', R'' are each independently selected from hydrogen, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl and C 1-3 Halogenated alkoxy.

[0068] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted C 1~5 Alkyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted 5- to 6-membered heteroaryl and substituted or unsubstituted 4- to 6-membered heterocyclic group, wherein the heteroaryl group has 1 to 2 heteroatoms selected from N, O, and S, and the heterocyclic group has 1 to 2 heteroatoms selected from N, O, and S.

[0069] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isopentyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperazinyl, and substituted or unsubstituted thiomorpholinyl.

[0070] In certain embodiments, R is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl, substituted or unsubstituted phenyl, substituted or unsubstituted furanyl, substituted or unsubstituted oxetanyl, substituted or unsubstituted azetidinyl, and substituted or unsubstituted piperidinyl.

[0071] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), C 1~6 Alkyl and C 1~6 Alkoxy.

[0072] In certain embodiments, each occurrence of Q is independently selected from hydrogen, deuterium, halogen, hydroxyl, NR'(R"), and C 1~3 alkyl.

[0073] In certain embodiments, each occurrence of Q is independently selected from hydroxy, NR'(R"), and methyl.

[0074] In certain embodiments, R' and R'' are each independently selected from hydrogen and C 1-3 alkyl.

[0075] In certain embodiments, R' and R" are each independently selected from hydrogen and methyl.

[0076] In certain embodiments, the present invention provides a compound or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug, and stable isotope derivative thereof, characterized in that the compound is any one of the following:

[0077] 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and .

[0078] In certain embodiments, the present invention provides a compound or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug, and stable isotope derivative thereof, characterized in that the compound is any one of the following:

[0079] 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and .

[0080] The present invention provides a pharmaceutical composition comprising the aforementioned compound or its pharmaceutically acceptable salts, tautomers, stereoisomers, solvates, hydrates, prodrugs and stable isotope derivatives.

[0081] In certain embodiments, the composition further comprises a pharmaceutically acceptable carrier, diluent or excipient.

[0082] In certain embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

[0083] In certain embodiments, the pharmaceutical composition comprises 0.01% to 99.99% of the aforementioned compound, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition comprises 0.1% to 99.9% of the aforementioned compound. In certain embodiments, the pharmaceutical composition comprises 0.5% to 99.5% of the aforementioned compound. In certain embodiments, the pharmaceutical composition comprises 1% to 99% of the aforementioned compound. In certain embodiments, the pharmaceutical composition comprises 2% to 98% of the aforementioned compound.

[0084] In certain embodiments, the pharmaceutical composition contains 0.01%-99.99% of a pharmaceutically acceptable carrier, diluent, or excipient, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1%-99.9% of a pharmaceutically acceptable carrier, diluent, or excipient. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of a pharmaceutically acceptable carrier, diluent, or excipient. In certain embodiments, the pharmaceutical composition contains 1%-99% of a pharmaceutically acceptable carrier, diluent, or excipient. In certain embodiments, the pharmaceutical composition contains 2%-98% of a pharmaceutically acceptable carrier, diluent, or excipient.

[0085] All compounds of the present invention, as well as mixtures and compositions comprising the compounds of the present invention, can be administered to a living organism via any route of administration. Routes of administration include oral administration, intravenous injection, intramuscular injection, subcutaneous injection, rectal administration, vaginal administration, sublingual administration, nasal inhalation, oral inhalation, eye drops, and topical or systemic transdermal administration.

[0086] All compounds of the present invention and mixtures and compositions containing the compounds of the present invention can be formulated into single doses, which contain the active compounds of the present invention as well as carriers, excipients, etc. The dosage forms can be tablets, capsules, injections, granules, powders, suppositories, pills, creams, pastes, gels, powders, oral solutions, inhalants, suspensions, dry suspensions, patches, lotions, etc. These dosage forms may contain ingredients commonly used in pharmaceutical preparations, such as diluents, absorbents, wetting agents, binders, disintegrants, colorants, pH adjusters, antioxidants, antibacterial agents, isotonicity adjusters, anti-adherents, etc.

[0087] Suitable formulations of the above-mentioned various dosage forms are available from public sources, such as Remington: The Science and Practice of Pharmacy, 21st edition, published by Lippincott Williams & Wilkins in 2006 and Rowe, Raymond C. Handbook of Pharmaceutical Excipients, Chicago, Pharmaceutical Press in 2005, and thus can be easily prepared by those skilled in the art.

[0088] Different dosages can be selected based on the nature and intensity of the disease suffered by different individuals, the patient's age, gender, weight, route of administration and other factors. The dosage of the compound of the present invention can be 0.01 to 500 mg / kg per day, preferably 1-100 mg / kg per day, and can be administered in single or multiple doses.

[0089] The present invention provides the use of the aforementioned compound or its pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, prodrug and stable isotope derivative, or the aforementioned pharmaceutical composition in preventing and / or treating GSPT1-related diseases.

[0090] The present invention provides use of the aforementioned compound or its pharmaceutically acceptable salts, tautomers, stereoisomers, solvates, hydrates, prodrugs and stable isotope derivatives, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating GSPT1-related diseases.

[0091] In certain embodiments, the GSPT1-associated disease is selected from cancer, autoimmune disease, immunodeficiency disease, aging, viral infection, and organ transplant rejection.

[0092] In certain embodiments, the cancer is selected from bladder cancer, breast cancer, bone cancer, neuronal cell cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, choriocarcinoma, multiple myeloma, basal cell carcinoma, teratoma, choroidal melanoma, seminoma, craniopharyngioma, plasmacytoma, papilloma, budding glioma, sarcoma (including but not limited to chondrosarcoma, histiocytoma, malignant fibrous histiocytoma, lymphosarcoma and rhabdomyosarcoma), melanoma, hemangioma, keloid, squamous cell carcinoma, astrocytoma, lymphoma (including but not limited to choroidal fibrosis), leukemia, myeloma, leukemia, sarcoma, leukemia, necrosis factor alpha ... Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, adult T-cell leukemia and lymphoma, diffuse large B-cell lymphoma, Hodgkin's disease, and central nervous system lymphoma), respiratory tract cancer (including but not limited to lung cancer, such as small cell lung cancer, non-small cell lung cancer, bronchial adenoma and thoracic pulmonary blastoma), head and neck cancer (including but not limited to head cancer, neck cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer and / or oropharyngeal cancer, and cancer of the lips and oral cavity), breast cancer (including but not limited to invasive ductal carcinoma, lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma), digestive tract cancer (including including but not limited to anal cancer, colon cancer, colorectal cancer, esophageal cancer, gallbladder cancer, rectal cancer, stomach cancer, small intestine cancer and salivary gland cancer), thyroid cancer, parathyroid cancer and their distant metastases, liver cancer (including but not limited to hepatocellular carcinoma, stem cell carcinoma with or without fibrolamellar form, bile duct cell carcinoma and mixed hepatocellular and cholangiocellular carcinoma), leukemia (including but not limited to acute lymphoblastic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia and villous cell leukemia), brain cancer (including but not limited to Brainstem and pituitary gliomas, medulloblastomas, cerebellar and cerebral astrocytomas, ependymoma and neuroectodermal tumors and pineal adenomas), reproductive organ cancers (including but not limited to prostate cancer, testicular cancer, ovarian cancer, endometrial cancer, cervical cancer, vaginal cancer and vulvar cancer, and uterine sarcoma), urethral cancer, eye cancer (including but not limited to intraocular melanoma and retinoblastoma), skin cancer (including but not limited to Kaposi's sarcoma, squamous cell carcinoma, malignant melanoma, Merkel cell skin cancer and non-melanoma skin cancer), renal parenchymal cancer, kidney cancer and other related cancers.

[0093] In certain embodiments, the virus in the viral infection is selected from RNA viruses.

[0094] In certain embodiments, the virus is selected from the group consisting of Coronaviridae, Orthomyxoviridae, Retroviridae, Flaviviridae, Poliovirus, Rabies, Poliovirus, Lassa virus, and Ebola virus.

[0095] Explanation of terms:

[0096] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0097] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of D atoms, halogen, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0098] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, preferably 3 to 8 (e.g., 3, 4, 5, 6, 7, and 8) carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls.

[0099] The term " spiroalkyl " refers to a polycyclic group of 5 to 20 yuan, a carbon atom (called spiral atom) shared between the monocycle, which can contain one or more double bonds. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of spiral atoms shared between the rings, spiroalkyl is divided into single spiroalkyl, double spiroalkyl or multiple spiroalkyl, preferably single spiroalkyl and double spiroalkyl. More preferably, it is 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiroalkyl. Non-limiting examples of spiroalkyl include:

[0100] .

[0101] The term "fused cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system, wherein one or more rings may contain one or more double bonds. Preferably, it is 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9, or 10 members). According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl groups, preferably bicyclic or tricyclic, more preferably 3 / 4, 3 / 5, 3 / 6, 4 / 4, 4 / 5, 4 / 6, 5 / 4, 5 / 5, 5 / 6, 6 / 3, 6 / 4, 6 / 5, and 6 / 6 bicyclic alkyl groups. Non-limiting examples of fused cycloalkyl groups include:

[0102] .

[0103] The term "bridged cycloalkyl" refers to a 5- to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, and which may contain one or more double bonds. Preferably, it is 6- to 14-membered, and more preferably, it is 7- to 10-membered (e.g., 7, 8, 9, or 10-membered). Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl group, preferably a bicyclic, tricyclic, or tetracyclic group, and more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged cycloalkyl groups include:

[0104]

[0105] The cycloalkyl ring includes a cycloalkyl group as described above (including monocyclic, spirocyclic, fused and bridged rings) fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples include 、 、 etc.; preferred and .

[0106] The cycloalkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0107] The term "alkoxy" refers to -O-(alkyl) and -O-(cycloalkyl), wherein alkyl and cycloalkyl are as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, and butoxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, are preferably one or more of the following groups independently selected from the group consisting of D atoms, halogen, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0108] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic non-aromatic cyclic substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen and sulfur, wherein the sulfur may be optionally oxoed (i.e., forming a sulfoxide or sulfone), but excluding the ring moiety -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, the substituent contains 3 to 12 ring atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12), of which 1 to 4 (e.g., 1, 2, 3 and 4) are heteroatoms; more preferably, the substituent contains 3 to 8 ring atoms (e.g., 3, 4, 5, 6, 7 and 8), of which 1 to 3 (e.g., 1, 2 and 3) are heteroatoms; more preferably, the substituent contains 3 to 6 ring atoms, of which 1 to 3 are heteroatoms; and most preferably, the substituent contains 5 or 6 ring atoms, of which 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclyls include spiro, fused, and bridged heterocyclyls.

[0109] The term "spiroheterocyclyl" refers to a 5-20-membered, non-aromatic polycyclic heterocyclic group in which one atom (called a spiro atom) is shared between the monocyclic rings, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur being optionally oxoed (i.e., forming a sulfoxide or sulfone), and the remaining ring atoms being carbon. It may contain one or more double bonds. It is preferably 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members). Spiroheterocyclyl is divided into monospiroheterocyclyl, bispiroheterocyclyl or polyspiroheterocyclyl according to the number of spiro atoms shared between the rings, preferably monospiroheterocyclyl and bispiroheterocyclyl. More preferably, it is a 3-membered / 5-membered, 3-membered / 6-membered, 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered or 5-membered / 6-membered monospiroheterocyclyl. Non-limiting examples of spiroheterocyclyl include:

[0110] .

[0111] The term "fused heterocyclic group" refers to a non-aromatic polycyclic heterocyclic group having 5 to 20 members, wherein each ring in the system shares a pair of adjacent atoms with other rings in the system. One or more rings may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur, wherein the sulfur may be optionally oxoed (i.e., forming a sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members (e.g., 7, 8, 9, or 10 members). Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic group, and more preferably a 3 / 4-membered, 3 / 5-membered, 3 / 6-membered, 4 / 4-membered, 4 / 5-membered, 4 / 6-membered, 5 / 4-membered, 5 / 5-membered, 5 / 6-membered, 6 / 3-membered, 6 / 4-membered, 6 / 5-membered, and 6 / 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

[0112]

[0113] and .

[0114] The term "bridged heterocyclic group" refers to a non-aromatic polycyclic heterocyclic group with 5 to 14 members, wherein any two rings share two atoms that are not directly connected. It may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur may be optionally oxoed (i.e., forming a sulfoxide or sulfone), and the remaining ring atoms are carbon. It is preferably 6 to 14 members, and more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members). Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, preferably a bicyclic, tricyclic or tetracyclic group, and more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged heterocyclic groups include:

[0115] .

[0116] The heterocyclyl ring includes a heterocyclyl as described above (including monocyclic, spiro heterocyclic, fused heterocyclic and bridged heterocyclic rings) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0117] and wait.

[0118] The heterocyclyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0119] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is a ring that shares adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aryl ring includes an aryl ring as described above fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

[0120] and .

[0121] Aryl can be substituted or unsubstituted, and when substituted, it can be substituted on any available point of attachment, and the substituent is preferably independently optionally selected from one or more substituents in halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 (e.g., 1, 2, 3, and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl is preferably 5 to 10 yuan (e.g., 5, 6, 7, 8, 9, or 10 yuan), more preferably 5 yuan or 6 yuan, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc. The heteroaryl ring includes a heteroaryl group as described above fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0122] and .

[0123] The heteroaryl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0124] The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0125] The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0126] The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, wherein alkyl is as defined above.

[0127] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0128] The term "hydroxy" refers to -OH.

[0129] The term "cyano" refers to -CN.

[0130] The compounds of the present disclosure include isotopic derivatives thereof. The term "isotopic derivative" refers to a compound whose structure differs only in the presence of one or more isotopically enriched atoms. For example, a compound having a structure of the present disclosure, with "deuterium" or "tritium" replacing hydrogen, or with 18 F-fluorine labeling ( 18 F isotope) instead of fluorine, or with 11 C-, 13 C-, or 14 C-enriched carbon ( 11 C-, 13 C-, or 14 C-carbon labeling; 11 C-, 13 C-, or 14 Compounds in which a carbon atom is replaced by a deuterium atom (C-isotope) are within the scope of this disclosure. Such compounds can be used, for example, as analytical tools or probes in biological assays, or as in vivo diagnostic imaging tracers for diseases, or as tracers for pharmacodynamic, pharmacokinetic, or receptor studies. The various deuterated forms of the compounds disclosed herein refer to compounds in which each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art will be able to synthesize deuterated forms of the compounds by referring to relevant literature. Deuterated forms of the compounds can be prepared using commercially available deuterated starting materials, or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated ethyl iodide, and deuterated methyl iodide. Deuterated compounds generally retain activity comparable to the undeuterated compounds, and deuteration at certain specific sites can achieve improved metabolic stability, thereby conferring certain therapeutic advantages.

[0131] The term "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0132] The term "substituted" refers to a group in which one or more hydrogen atoms, preferably 1 to 5, more preferably 1 to 3, are independently replaced by a corresponding number of substituents. Those skilled in the art will be able to determine (by experiment or theory) whether substitution is possible or not without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0133] The term "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0134] The term "pharmaceutically acceptable salt" refers to salts of the disclosed compounds that are safe and effective for use in mammals and possess the desired biological activity. Salts can be prepared during the final isolation and purification of the compound, or separately by reacting a suitable group with a suitable base or acid. Bases commonly used to form pharmaceutically acceptable salts include inorganic bases, such as sodium hydroxide and potassium hydroxide, and organic bases, such as ammonia. Acids commonly used to form pharmaceutically acceptable salts include inorganic acids and organic acids.

[0135] With respect to a drug or pharmacologically active agent, the term "therapeutically effective amount" refers to a non-toxic amount of the drug or agent sufficient to achieve the desired effect. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active agent. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0136] The term "solvate" refers to a physical association of a compound of the present disclosure with one or more, preferably 1-3, solvent molecules, whether organic or inorganic. This physical association includes hydrogen bonding. In certain cases, for example, when one or more, preferably 1-3, solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be isolated. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are well known in the art.

[0137] The term "hydrate" refers to a substance formed by the compound of the present application or a pharmaceutically acceptable salt thereof and water through non-covalent intermolecular forces. Common hydrates include (but are not limited to) hemihydrates, monohydrates, dihydrates, trihydrates, etc.

[0138] The term "prodrug" refers to a compound that can be transformed in vivo under physiological conditions, for example, by hydrolysis in the blood, to yield the active prodrug.

[0139] The term "stereoisomers" refers to compounds that have the same chemical constitution but differ in the way the atoms or groups are arranged in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric (cis / trans) isomers, atropisomers, and the like.

[0140] The term "tautomer" refers to isomers of a compound that differ from one another in the location of protons and / or distribution of electrons. Examples of tautomers include, but are not limited to, enol-ketone tautomers, imine-enamine tautomers, amide-imidic acid tautomers, amine-imine tautomers, and tautomeric forms of heteroaryls comprising ring atoms connected to the -NH- portion of the ring and the =N- portion of the ring, such as pyrazole, imidazole, benzimidazole, triazole, pyridotriazole, piperidotriazole, and tetrazole.

[0141] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with patient tissues without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio, and effective for the intended use.

[0142] The term "deuterated" refers to a compound in which one or more hydrogen atoms are replaced by deuterium atoms. Deuteration can be mono-, di-, poly- or full-substituted. The term "deuterated compound" refers to a compound containing deuterium atoms.

[0143] As used herein, the singular form "a," "an," and "the" include plural references and vice versa unless the context clearly dictates otherwise.

[0144] In the chemical structure of the compound of the present invention, the bond " " indicates that the configuration is not specified, i.e. if chiral isomers exist in the chemical structure, the bond " ” can be "or" ”, or both "and" "Two configurations. Key" ” can be ”, “ " is a cis structure. Although all the above structural formulas are drawn as certain isomers for the sake of simplicity, the present invention can include all isomers, such as tautomers, rotational isomers, geometric isomers, diastereomers, racemates and enantiomers. In the chemical structure of the compounds disclosed in the present invention, the bond " " does not specify the configuration, i.e. the bond" The configuration of " can be E-type or Z-type, or contain both E and Z configurations.

[0145] The terms "administering" or "administering" refer to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound described herein or a composition thereof into the body or onto a subject.

[0146] The term "treating" includes any effect that results in improvement, such as alleviation, reduction, regulation, amelioration or elimination, of a condition, disease, disorder, etc., or amelioration of the symptoms thereof.

[0147] The term "cancer," as used herein, refers to an abnormal, uncontrolled growth of cells that, under certain conditions, can metastasize. This type of cancer includes, but is not limited to, solid tumors (e.g., of the bladder, intestine, brain, breast, uterus, heart, kidney, lung, lymphoid tissue, ovary, pancreas or other endocrine organs (e.g., thyroid), prostate, and skin (melanoma)) or blood tumors (e.g., non-leukemic leukemias).

[0148] In the present invention, cancer includes bladder cancer, breast cancer, bone cancer, neuronal cell cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, choriocarcinoma, multiple myeloma, basal cell carcinoma, teratoma, choroidal melanoma, seminoma, craniopharyngioma, plasmacytoma, papilloma, budding glioma, sarcoma melanoma, hemangioma, keloid, squamous cell carcinoma, astrocytoma, lymphoma, respiratory tract cancer, head and neck cancer, breast cancer, digestive tract cancer, thyroid cancer, parathyroid cancer and its distant metastases, liver cancer, leukemia, brain cancer, reproductive organ cancer, urethra cancer, eye cancer, skin cancer, renal parenchymal cancer, kidney cancer.

[0149] The sarcoma includes but is not limited to chondrosarcoma, histiosarcoma, malignant fibrous histiocytoma, lymphosarcoma and rhabdomyosarcoma. In certain embodiments, the lymphoma includes but is not limited to Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma, Hodgkin's disease and central nervous system lymphoma. In certain embodiments, the respiratory tract cancer includes but is not limited to lung cancer, such as small cell, non-small cell lung cancer, and bronchial adenoma and thoracic pulmonary blastoma. In certain embodiments, the head and neck cancer includes but is not limited to head cancer, neck cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer and / or oropharyngeal cancer and lip and oral cancer. In certain embodiments, the breast cancer includes but is not limited to invasive ductal carcinoma, lobular carcinoma, ductal carcinoma in situ and lobular carcinoma. In certain embodiments, the digestive tract cancer includes but is not limited to anal cancer, colon cancer, colorectal cancer, esophageal cancer, gallbladder cancer, rectal cancer, gastric cancer, small intestine cancer and salivary gland cancer. In certain embodiments, the liver cancer includes but is not limited to hepatocellular carcinoma, stem cell cancer with or without fibrolamellar form, cholangiocarcinoma and mixed hepatocellular cholangiocarcinoma. In certain embodiments, the leukemia includes but is not limited to acute lymphoblastic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia and chorionic villus leukemia. In certain embodiments, brain cancer includes but is not limited to brainstem and pituitary glioma, medulloblastoma, cerebellar and cerebral astrocytoma, ependymoma and neuroectodermal tumor and pineal adenoma. In certain embodiments, reproductive organ cancer includes but is not limited to prostate cancer, testicular cancer, ovarian cancer, endometrial cancer, cervical cancer, vaginal cancer and vulvar cancer and uterine sarcoma, in certain embodiments, eye cancer includes but is not limited to intraocular melanoma and retinoblastoma, in certain embodiments, skin cancer includes but is not limited to Kaposi's sarcoma, squamous cell tumor, malignant melanoma, Merkel cell skin cancer and non-melanoma skin cancer.

[0150] The term "virus", as used herein, refers to a non-cellular organism that is tiny, simple in structure, contains only one nucleic acid (DNA or RNA), and must parasitize and replicate within living cells. These viruses include, but are not limited to, rhinovirus, poliovirus, rotavirus, norovirus, enterovirus, hepatovirus, astrovirus, sapovirus, hepatitis E virus, influenza A / B / C virus, parainfluenza virus, mumps virus, measles virus, human metapneumovirus, RS virus, Nipah virus, Hendra virus, yellow fever virus, dengue virus, Japanese encephalitis virus, West Nile virus, hepatitis B / C virus, eastern and western equine encephalitis virus, Arnold-Nyon virus, rubella virus, Lassa virus, Junin virus, Maqiu virus, and other viruses. Bo virus, Guanarito virus, arenavirus, Crimean-Congo hemorrhagic fever virus, sandfly fever virus, Hantavirus, Sin Nombre virus, rabies virus, Ebola virus, Marburg virus, bat lyssavirus, human T-cell leukemia virus, human immunodeficiency virus, human coronavirus, SARS coronavirus, human parvovirus, human polyomavirus, human papillomavirus, adenovirus, herpes virus, varicella virus, herpes zoster virus, Epstein-Barr virus, cytomegalovirus, smallpox virus, monkeypox virus, cowpox virus, molluscum pox virus and parapox virus.

[0151] The above definitions are given for the terms involved in the present invention. Those skilled in the art can also understand the above terms in combination with the existing technology. The following is a further description based on the content of the present invention and the definitions of the terms.

[0152] The following examples further describe the preparation of the compounds and pharmaceutically acceptable salts of the present invention, but these examples are not intended to limit the scope of the present invention.

[0153] The experimental methods in the examples of the present invention, where specific conditions are not specified, are generally carried out under conventional conditions or the conditions recommended by the raw material or product manufacturers. Reagents where the specific sources are not specified are conventional reagents purchased from the market.

[0154] The following describes the general experimental conditions in the embodiments of the present invention:

[0155] First, the reactions in the examples are generally carried out under nitrogen protection.

[0156] Furthermore, the intermediates and final products were separated and purified by chromatographic columns, preparative chromatography plates and ISCO rapid preparative chromatography system.

[0157] Furthermore, the LC-MS liquid chromatography-mass spectrometry instrument uses Waters ACQUITY Arc or equivalent equipment. Mass spectrometry (MS) uses an ESI source and only indicates the molecular weight M of the parent molecule, usually reporting [M+H] +NMR spectra were obtained using a Varian 400 MHz NMR spectrometer or equivalent, typically using CDCl₃ or DMSO-d₆ as solvents, and chemical shifts are reported in ppm. Peaks are described as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), and dd (doublet of doublets). Coupling constants are expressed in Hz. Example 1

[0158] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxyphenyl)carbamate (Compound 1)

[0159]

[0160] Step 1: Preparation of phenyl (2-fluoro-5-trifluoromethoxyphenyl)carbamate

[0161]

[0162] 2-Fluoro-5-trifluoromethoxyaniline (507 mg, 2.6 mmol), phenyl chloroformate (564 mg, 3.6 mmol), and pyridine (403 mg, 5.1 mmol) were dissolved in acetonitrile (10 mL) and reacted at room temperature for 4 h. TLC monitored the reaction completion. The acetonitrile was removed under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. The organic phase was stripped of the solvent under reduced pressure, and the residue was purified by flash column chromatography (PE:EA = 80:1). 600 mg of the desired product was obtained in a 74% yield.

[0163] 1 H NMR (400 MHz, DMSO-d 6 ) d 9.31 (s, 1H), 7.50 – 7.36 (m, 2H), 7.17 –7.12(m, 2H), 7.06 (dd, J = 12.0, 8.0 Hz, 1H), 6.78 – 6.73 (m, 3H).

[0164] Step 2: Preparation of 4-methylbenzene-1,3-dicarboxylic acid

[0165]

[0166] Dissolve methyl 5-cyano-2-methylbenzoate (10.0 g, 57.1 mmol) in water (250 mL), then add concentrated sulfuric acid (100 mL). After addition, react at 100°C. TLC monitors the reaction for completion. Cool the mixture to room temperature, filter, and wash the filter cake with water and dry to constant weight. The desired product (10.0 g) is obtained in a 98% yield.

[0167] 1 H NMR (400 MHz, DMSO-d 6 ) d 8.37 (d, J = 2.0 Hz, 1H), 7.97 (dd, J =8.0, 2.0 Hz, 1H), 7.43 (d, J = 8.0 Hz, 1H), 2.58 (s, 3H).

[0168] Step 3: Preparation of 5-bromo-4-methylbenzene-1,3-dicarboxylic acid

[0169]

[0170] Dissolve 4-methylbenzene-1,3-dicarboxylic acid (10.0 g, 55.6 mmol) in concentrated sulfuric acid (60 mL), and add NBS (8.9 g, 50.0 mmol) portionwise. After addition, react at 50°C for 4 h. TLC monitors the reaction for completion. Cool the mixture to room temperature, then slowly pour it into ice water and stir for 10 min. Filter the mixture, wash the filter cake with water, anhydrous ethanol, and n-hexane, and dry the resulting solid to constant weight. This yields 14.1 g of the desired product in a 99% yield.

[0171] 1 H NMR (400 MHz, DMSO-d 6 ) d 8.18 (s, 1H), 8.14 (s, 1H), 2.56 (s, 3H).

[0172] Step 4: Preparation of methyl 5-bromo-4-methylbenzene-1,3-dicarboxylate

[0173]

[0174] Dissolve 5-bromo-4-methylbenzene-1,3-dicarboxylic acid (14.1 g, 54.7 mmol) in methanol (350 mL), then add concentrated sulfuric acid (3.0 mL). After addition, allow to react overnight at 65°C. TLC confirms the reaction is complete the next day. Cool the system to room temperature, remove the solvent under reduced pressure, and purify the residue by flash column chromatography (PE:EA = 20:1). The desired product (12.8 g) is obtained in an 82% yield.

[0175] 1 H NMR (400 MHz, CDCl 3 ) d 8.35 (dd, J = 16.0, 4.0 Hz, 2H), 3.92 (s, 6H), 2.67 (s, 3H).

[0176] Step 5: Preparation of methyl 5-bromo-4-bromomethylbenzene-1,3-dicarboxylate

[0177]

[0178] Methyl 3-bromo-5-(methoxycarbonyl)-4-methylbenzoate (12.8 g, 44.5 mmol), NBS (11.9 g, 66.7 mmol), and AIBN (739 mg, 4.5 mmol) were dissolved in CHCl₃ (200 mL) and refluxed at 65°C overnight. TLC confirmed the reaction completion the next day. After cooling, water was added, and the mixture was extracted with DCM. The organic phase was stripped of solvent under reduced pressure, and the residue was purified by flash column chromatography (PE:EA = 70:1). 14.0 g of the desired product was obtained in an 86% yield.

[0179] 1 H NMR (400 MHz, CDCl 3 ) d 8.50 (s, 1H), 8.38 (s, 1H), 5.13 (s, 2H), 3.97 (s, 3H), 3.94 (s, 3H).

[0180] Step 6: Preparation of methyl 7-bromo-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindole-5-carboxylate

[0181]

[0182] Methyl 5-bromo-4-bromomethylbenzene-1,3-dicarboxylate (12.6 g, 41.0 mmol), 3-amino-2,6-piperidinedione hydrochloride (10.1 g, 61.5 mmol), and DIPEA (15.9 g, 123.0 mmol) were dissolved in acetonitrile (200 mL) and reacted at 80°C overnight. TLC confirmed the reaction was complete the next day. After cooling, the mixture was filtered, and the filter cake was washed with acetonitrile and dried. The product was obtained as a gray-black solid (11.0 g, 83% yield).

[0183] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.03 (s, 1H), 8.33 (d, J = 1.0 Hz, 1H),8.20 (d, J = 1.0 Hz, 1H), 5.16 (dd, J = 12.0, 4.0 Hz, 1H), 4.53 (d, J =16.0Hz, 1H), 4.37 (d, J =20.0 Hz, 1H), 3.92 (s, 3H), 2.97-2.86 (m, 1H), 2.68-2.56 (m, 1H), 2.49 – 2.42 (m, 1H), 2.07-1.96 (m, 1H).

[0184] Step 7: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate

[0185]

[0186] Methyl 7-bromo-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindole-5-carboxylate (4.0 g, 10.5 mmol) was dissolved in anhydrous DMF (50 mL), followed by the addition of CuI (399 mg, 2.1 mmol), DIPEA (6.8 g, 52.5 mmol), and PdCl2(PPh3)2 (737 mg, 1.05 mmol). The nitrogen atmosphere was replaced, and trimethylsilylacetylene (1.6 g, 16.3 mmol) was added. After the addition was complete, the reaction was allowed to proceed at 60°C. TLC monitored the reaction completion. The system was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was stripped of solvent under reduced pressure, and the residue was purified by flash column chromatography (DCM:MeOH = 90:1). 3.5 g of the desired product was obtained in an 80% yield.

[0187] 1 H NMR (400 MHz, CDCl 3 ) d 8.48 (d, J = 1.0 Hz, 1H), 8.33 (d, J = 1.0Hz, 1H), 8.17 (s, 1H), 5.26 (dd, J = 12.0, 4.0 Hz, 1H), 4.56 (d, J = 16.0 Hz,1H), 4.40 (d, J = 16.0 Hz, 1H), 3.96 (s, 3H), 2.99 – 2.80 (m, 3H), 2.50-2.36(m, 1H), 2.30-2.18 (m, 1H), 1.63 (s, 1H), 0.27 (s, 9H).

[0188] Step 8: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole

[0189]

[0190] Methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate (2.9 g, 7.2 mmol) was dissolved in anhydrous THF (100 mL), and LiAlH4 (547 mg, 14.4 mmol) was added under ice-cooling. After the addition was complete, the reaction was allowed to warm naturally. TLC monitored the reaction completion. The solvent was removed under reduced pressure, and water was added to the residue. The pH was adjusted to 1-2 with 2M aqueous HCl. The product was extracted with ethyl acetate, and the organic phase was stripped of the solvent under reduced pressure. The residue was purified by flash column chromatography (DCM:MeOH = 40:1). 215 mg of the desired product was obtained in a 10% yield.

[0191] 1 H NMR (400 MHz, CDCl 3 ) d 7.75 (s, 1H), 7.62 (s, 1H), 5.09 (dd, J =12.0, 4.0 Hz, 1H), 4.64 (s, 2H), 4.48 – 4.29 (m, 2H), 3.29 (t, J= 1.6 Hz,1H), 2.86 – 2.69 (m, 2H), 2.39-2.25 (m, 1H), 2.18 – 2.10 (m, 1H).

[0192] Step 9: Preparation of Compound 1

[0193]

[0194] (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole (105 mg, 0.3 mmol) was dissolved in DMF (4 mL). NaH (24 mg, 0.6 mmol) was added under ice-cooling, followed by a DMF solution of phenyl (2-fluoro-5-trifluoromethoxyphenyl)carbamate (98 mg, 0.3 mmol). The reaction was allowed to warm naturally for 3 h. TLC monitored the reaction completion. Aqueous ammonium chloride was added to the system to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with brine. The solvent was removed under reduced pressure, and the residue was purified by flash column chromatography (DCM:MeOH = 40:1). 52 mg of the desired product was obtained in a 30% yield.

[0195] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.00 (s, 1H), 9.96 (s, 1H), 7.90 – 7.80 (m, 3H), 7.39 (m, 1H), 7.20 – 7.09 (m, 1H), 5.30 (s, 2H), 5.15 (dd, J = 12.0,4.0 Hz, 1H), 4.65 (s, 1H), 4.51 (d, J = 20.0 Hz, 1H), 4.35 (d, J = 20.0 Hz,1H), 2.98 – 2.84 (m, 1H), 2.64 – 2.54 (m, 1H), 2.48-2.41 (m, 1H), 2.03-2.0(m, 1H).

[0196] MS-ESI: 520.1[M+H] + . Example 2

[0197] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)methyl(2-fluoro-5-(3-hydroxy-3-methylbut-1-yn-1-yl)phenyl)carbamate (Compound 2)

[0198]

[0199] Step 1: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(prop-1-ynyl)isoindole-5-carboxylate

[0200]

[0201] Methyl 7-bromo-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindole-5-carboxylate (4.0 g, 10.5 mmol), (tributyltin)propyne (4.14 mg, 12.6 mmol), and Pd(PPh3)4 (1.21 mg, 1.1 mmol) were dissolved in toluene (100 mL), the atmosphere was replaced with nitrogen, and the mixture was reacted at 115°C overnight. The next day, TLC confirmed the completion of the reaction. The system was cooled, and aqueous KF solution (5.0 g, 40 mL) was added and stirred for 30 min. A 10:1 mixture of DCM and MeOH was then added and stirred for 10 min. The mixture was filtered through celite. The solvent was removed from the filtrate under reduced pressure, and the product was extracted with ethyl acetate. The solvent was removed from the organic phase under reduced pressure, and the residue was purified by flash column chromatography (DCM:MeOH = 90:1). 2.28 g of the desired product was obtained in a yield of 64%.

[0202] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.02 (s, 1H), 8.13 (s, 1H), 8.11 (s, 1H),5.16 (dd, J = 12.0, 4.0 Hz, 1H), 4.48 (dd, J = 64.0, 20.0 Hz, 2H), 3.90 (s,3H), 2.97 – 2.85 (m, 1H), 2.67 – 2.55 (m, 1H), 2.51 – 2.42 (m, 1H), 2.13 (s,3H), 2.06 – 1.95 (m, 1H).

[0203] Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(prop-1-ynyl)-isoindole

[0204]

[0205] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(prop-1-ynyl)-isoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(prop-1-ynyl)-isoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(prop-1-ynyl)isoindole-5-carboxylic acid methyl ester. 230 mg of the target product was obtained in a yield of 11%.

[0206] 1 H NMR (400 MHz, CDCl 3 ) d 8.16 (s, 1H), 7.77 (s, 1H), 7.58 (s, 1H),5.21 (dd, J = 12.0, 4.0 Hz, 1H), 4.75 (s, 2H), 4.40 (dd, J = 56.0, 16.0 Hz,1H), 2.96 – 2.76 (m, 2H), 2.43 – 2.32 (m, 1H), 2.24 – 2.19 (m, 1H), 2.09 (s,3H).

[0207] Step 3: Preparation of Compound 2

[0208]

[0209] Compound 2 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-propynylisoindole). 83 mg of the target product was obtained in a yield of 49%.

[0210] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.00 (s, 1H), 9.94 (s, 1H), 7.85 (d, J =4.0 Hz, 1H), 7.79 (s, 1H), 7.71 (s, 1H), 7.39 (dd, J= 12.0, 8.0 Hz, 1H),7.15 – 7.12 (m, 1H), 5.28 (s, 2H), 5.15 (dd, J = 12.0, 4.0 Hz, 1H), 4.40 (dd, J = 64.0, 20.0 Hz, 2H), 2.99 – 2.85 (m, 1H), 2.66 – 2.57 (m, 1H), 2.49 – 2.40(m, 1H), 2.12 (s, 3H), 2.04 – 1.98 (m, 1H).

[0211] MS-ESI: 534.1 [M+H] + . Example 3

[0212] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopropylethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 3)

[0213]

[0214] Step 1: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopropylethynylisoindole-5-carboxylate

[0215]

[0216] The synthesis of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopropylethynylisoindole-5-carboxylate) was the same as that of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate) except that trimethylsilyl acetylene was replaced with ethynylcyclopropane (260 mg, 3.9 mmol). The target product was obtained in a yield of 74% (710 mg).

[0217] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.01 (s, 1H), 8.12 (d, J = 2.0 Hz, 1H),8.09 (d, J = 2.0 Hz, 1H), 5.15 (dd, J = 12.0, 4.0 Hz, 1H), 4.46 (dd, J=60.0, 20.0 Hz, 2H), 3.90 (s, 3H), 2.96 – 2.87 (m, 1H), 2.67 – 2.57 (m, 1H), 2.54 – 2.43 (m, 1H), 2.05 – 1.96 (m, 1H), 1.65 – 1.58 (m, 1H), 0.99 – 0.93 (m, 2H), 0.88 – 0.81 (m, 2H).

[0218] Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopropylethynylisoindole

[0219]

[0220] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopropylethynylisoindole was the same as (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopropylethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopropylethynylisoindole-5-carboxylic acid methyl ester). 114 mg of the target product was obtained in a yield of 13%.

[0221] 1 H NMR (400 MHz, CDCl 3 ) d 8.00 (s, 1H), 7.77 (s, 1H), 7.58 (s, 1H), 5.23 (dd, J = 12.0, 4.0 Hz, 1H), 4.75 (s, 2H), 4.39 (dd, J = 56.0, 16.0 Hz,2H), 2.99 – 2.78 (m, 2H), 2.44 – 2.34 (m, 1H), 2.28 – 2.17 (m, 1H), 1.52 –1.42 (m, 1H), 0.96 – 0.90 (m, 2H), 0.84 – 0.78 (m, 2H).

[0222] Step 3: Preparation of Compound 3

[0223]

[0224] Compound 3 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopropylethynylisoindole). 48 mg of the target product was obtained in a yield of 26%.

[0225] 1 H NMR (400 MHz, DMSO-d 6 ) d 10.99 (s, 1H), 9.93 (s, 1H), 7.84 (d, J =4.0 Hz, 1H), 7.77 (s, 1H), 7.68 (s, 1H), 7.43 – 7.35 (m, 1H), 7.17 – 7.10 (m,1H), 5.27 (s, 2H), 5.14 (dd, J = 12.0, 4.0 Hz, 1H), 4.37 (dd, J = 56.0, 16.0Hz, 2H), 2.96 – 2.87 (m, 1H), 2.67 – 2.55 (m, 1H), 2.53 – 2.42 (m, 1H), 2.05– 1.96 (m, 1H), 1.65 – 1.56 (m, 1H), 0.97 – 0.91 (m, 2H), 0.83 – 0.78 (m, 2H).

[0226] MS-ESI: 560.1 [M+H] + . Example 4

[0227] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclobutylethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 4)

[0228]

[0229] Step 1: Preparation of ethynylcyclobutane

[0230]

[0231] Dissolve cyclobutanaldehyde (1.0 g, 11.9 mmol) in methanol (30 mL). Add dimethyl (1-diazo-2-oxopropyl)phosphonate (2.9 g, 15.5 mmol) and potassium carbonate (4.1 g, 29.7 mmol) sequentially under an ice bath. After addition, react at room temperature for 5 h. TLC monitors the reaction for completion. Add water (100 mL) to the system, extract with dichloromethane (20 mL), and wash the organic phase with water (100 mL x 2), dry over anhydrous MgSO₄, and use directly in the next step.

[0232] Step 2: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclobutylethynylisoindole-5-carboxylate

[0233]

[0234] The synthesis of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclobutylethynylisoindole-5-carboxylate) was similar to that of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate) except that trimethylsilyl acetylene was replaced with ethynylcyclobutane. 687 mg of the target product was obtained in a yield of 78%.

[0235] 1 H NMR (400 MHz, CDCl 3 ) d 8.44 (d, J = 1.5 Hz, 1H), 8.27 (d, J = 1.4Hz, 1H), 7.96 (s, 1H), 5.26 (dd, J = 13.4, 5.1 Hz, 1H), 4.46 (dd, J = 66.1,17.3 Hz, 2H), 3.96 (s, 3H), 3.26 (p, J = 8.4 Hz, 1H), 2.88 (m, 2H), 2.39 (m,3H), 2.23 (m, 3H), 2.01 (m, 2H).

[0236] Step 3: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclobutaneethynylisoindole

[0237]

[0238] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclobutylethynylisoindole) was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclobutylethynylisoindole), except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclobutylethynylisoindole-5-carboxylic acid methyl ester. 180 mg of the target product was obtained in a yield of 28%.

[0239] 1 H NMR (400 MHz, CDCl 3 ) d 8.44 (s, 1H), 7.74 (s, 1H), 7.58 (s, 1H), 5.20 (dd, J = 13.4, 5.0 Hz, 2H), 4.74 (t, J = 5.4 Hz, 3H), 4.36 (dd, J =53.7, 16.7 Hz, 3H), 3.25 (dd, J = 16.8, 8.4 Hz, 1H), 2.85 (m, 3H), 2.70 (d, J = 6.2 Hz, 1H), 2.35 (m, 4H), 2.21 (m, 4H), 1.96 (m, 3H).

[0240] Step 4: Preparation of Compound 4

[0241]

[0242] Compound 4 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclobutaneethynylisoindole). 43 mg of the target product was obtained in a yield of 29%.

[0243] 1 H NMR (400 MHz, CDCl 3 ) d 8.15 (s, 1H), 8.10 (s, 1H), 7.83 (d, J = 1.5Hz, 1H), 7.61 (d, J= 1.5 Hz, 1H), 7.08 (m, 3H), 6.87 (m, 1H), 5.28 (s, 2H), 5.24 (dd, J = 13.2, 5.0 Hz, 2H), 4.41 (dd, J = 61.4, 16.8 Hz, 3H), 3.25 (m,2H), 2.88 (m, 2H), 2.37 (m, 4H), 2.22 (m, 4H), 1.98 (m, 3H).

[0244] MS-ESI: 572.2 [MH] - . Example 5

[0245] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-oxetanyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 5)

[0246]

[0247] Step 1: Preparation of 3-ethynyloxetane

[0248]

[0249] 3-Ethynyloxetane was synthesized into ethynylcyclobutane except that cyclobutanaldehyde was replaced by oxetane-3-carboxaldehyde.

[0250] Step 2: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-oxetanyl)ethynylisoindole-5-carboxylate

[0251]

[0252] The synthesis of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-oxetanyl)ethynylisoindole-5-carboxylate was similar to that of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate), except that trimethylsilyl acetylene was replaced with 3-ethynyloxetane. 466 mg of the target product was obtained in a yield of 58%.

[0253] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.02 (s, 1H), 8.18 (q, J= 1.5 Hz, 2H),5.17 (dd, J = 13.3, 5.1 Hz, 1H), 4.83 (dd, J = 8.5, 5.4 Hz, 2H), 4.69 (m,2H), 4.53 (dd, J = 59.7, 18.7 Hz, 2H), 4.22 (tt, J = 8.5, 7.0 Hz, 1H), 3.91(s, 3H), 2.92 (m, 1H), 2.46 (dd, J = 13.2, 4.5 Hz, 1H), 2.02 (ddd, J = 20.1,9.5, 4.8 Hz, 2H).

[0254] Step 3: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-oxetanyl)ethynylisoindole

[0255]

[0256] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-oxetanyl)ethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-oxetanyl)ethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-oxetanyl)ethynylisoindole-5-carboxylic acid methyl ester. 175 mg of the target product was obtained in a yield of 13%.

[0257] 1 H NMR (400 MHz, CDCl 3 ) d 8.05 (s, 1H), 7.83 (s, 1H), 7.66 (s, 1H), 5.25 (dd, J = 13.3, 5.1 Hz, 1H), 4.91 (dd, J = 8.5, 5.5 Hz, 3H), 4.79 (dd, J = 7.0, 5.7 Hz, 6H), 4.43 (dd, J= 59.7, 16.6 Hz, 3H), 4.09 (m, 1H), 2.94 (d, J = 17.2 Hz, 1H), 2.85 (ddd, J = 17.9, 13.0, 5.3 Hz, 1H), 2.23 (dt, J = 11.3,5.3 Hz, 2H), 2.00 (m, 1H).

[0258] Step 4: Preparation of Compound 5

[0259]

[0260] Compound 5 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-oxetanyl)ethynylisoindole. 5 mg of the target product was obtained in a yield of 29%.

[0261] 1 H NMR (400 MHz, CDCl 3 ) d 8.10 (s, 1H), 7.96 (s, 1H), 7.89 (s, 1H),7.66 (d, J = 1.5 Hz, 1H), 7.09 (dd, J = 10.4, 9.0 Hz, 2H), 7.00 (m, 1H), 6.88(d, J = 8.9 Hz, 1H), 5.25 (dd, J = 13.4, 5.1 Hz, 1H), 4.92 (dd, J = 8.5, 5.5Hz, 3H), 4.79 (dd, J = 7.2, 5.5 Hz, 3H), 4.44 (dd, J = 63.3, 16.8 Hz, 3H), 4.10 (m, 1H), 2.87 (m, 3H), 2.40 (m, 2H), 2.23 (m, 3H), 2.01 (dd, J = 14.5,8.3 Hz, 3H).

[0262] MS-ESI: 574.4 [MH] -. Example 6

[0263] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-azetidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 6)

[0264]

[0265] Step 1: Preparation of tert-butyl 3-ethynyl-1-azetidinecarboxylate

[0266]

[0267] Ethynylcyclobutane was synthesized from tert-butyl 3-ethynyl-1-azetidinecarboxylate except that cyclobutanaldehyde was replaced by 1-Boc-azetidine-3-carbaldehyde.

[0268] Step 2: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindole-5-carboxylate

[0269]

[0270] The synthesis of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindole-5-carboxylate was the same as that of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate), except that trimethylsilyl acetylene was replaced with tert-butyl 3-ethynyl-1-azetidinecarboxylate. 2.1 g of the target product was obtained in a yield of 55%.

[0271] 1 H NMR (400 MHz, CDCl 3 ) d 8.49 (d, J = 4.0Hz, 1H), 8.29 (d, J = 4.0Hz,1H), 7.97 (s, 1H), 5.31 – 5.23 (dd, J =12.0,4.0 Hz,1H), 4.48 (dd, J = 24.0,16.0 Hz, 2H), 4.25 (t, J = 8.0 Hz, 2H), 4.02 (dd, J= 8.0, 8.0 Hz, 2H), 3.96(s, 3H), 3.57 (tt, J = 8.0, 4.0 Hz, 1H), 3.04 – 2.76 (m, 2H), 2.49 – 2.18 (m,2H), 1.45 (s, 9H).

[0272] Step 3: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-Boc-azetidinyl)ethynylisoindole

[0273]

[0274] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-Boc-azetidinyl)ethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-Boc-azetidinyl)ethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindole-5-carboxylic acid methyl ester. 297 mg of the target product was obtained with a yield of 15%.

[0275] MS-ESI: 454.4 [M+H] + .

[0276] Step 4: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate

[0277]

[0278] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindole was similar to that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(3-Boc-azetidinyl)ethynylisoindole was replaced with (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindole-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate. 100 mg of the target product was obtained in a yield of 23%.

[0279] 1 H NMR (400 MHz, CDCl 3 ) d 8.10 (s, 1H), 7.90 (d, J = 20.0 Hz, 2H),7.65 (s, 1H), 7.13–7.05 (m, 1H), 7.00 (d, J = 4.0 Hz, 1H), 6.88 (d, J = 8.0Hz, 1H), 5.30 (d, J = 4.0 Hz, 2H), 5.25 (dd, J = 16.0, 8.0 Hz, 1H), 4.44 (dd, J = 60.0, 16.0 Hz, 2H), 4.24 (t, J = 8.0 Hz, 2H), 4.01 (dd, J = 8.0, 8.0 Hz,2H), 3.65–3.50 (m, 1H), 3.05–2.78 (m, 2H), 2.43 (qd, J = 12.0, 4.0 Hz, 1H), 2.30–2.18 (m, 1H), 1.45 (s, 9H).

[0280] Step 5: Preparation of Compound 6

[0281]

[0282] (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (100 mg, 0.15 mmol) was dissolved in DCM (3 mL) and HCl / 1,4-dioxane (3 mL) was added. The reaction was allowed to react overnight at room temperature. The next day, the reaction was complete as monitored by TLC. The reaction solution was concentrated under reduced pressure, and the residue was slurried in anhydrous ethanol (3 mL). The resulting solid was collected by filtration and oven-dried. 38 mg of a white solid was obtained, with a yield of 40%.

[0283] 1 H NMR (400 MHz, DMSO-d 6 ) d11.02 (s, 1H), 9.94 (s, 1H), 9.30 (s, 1H),7.90 – 7.74 (m, 3H), 7.39 (t, J = 8.0 Hz, 1H), 7.17 – 7.10 (m, 1H), 5.31 (s,2H), 5.18 (dd, J = 16.0,8.0 Hz, 1H), 4.55 (d, J = 16 Hz, 1H), 4.42 – 4.34 (m,1H), 4.25 (t, J = 8.0 Hz, 2H), 4.10 – 3.93 (m, 3H), 3.57 (s, 1H), 2.62 (d, J = 16.0 Hz, 1H), 2.46 – 2.39 (m, 1H).

[0284] MS-ESI: 575.2 [M+H] + . Example 7

[0285] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-methylazetidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 7)

[0286]

[0287]

[0288] Compound 6 (38 mg, 0.06 mmol) was dissolved in methanol (3 mL), followed by the addition of 38% formaldehyde solution (8 mg, 0.08 mmol) and acetic acid (12 mg, 2 mmol). The reaction was allowed to react at room temperature for 2.5 h. NaBH3CN (12.5 mg, 0.2 mmol) was then added and allowed to react at room temperature for 3 h. Completion of the reaction was monitored by TLC. The reaction solution was concentrated under reduced pressure, adjusted to pH 14 by adding NaOH solution, diluted with water, and extracted with EA. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified using preparative chromatography to obtain 7 mg of a white solid, in an 18% yield.

[0289] 1 H NMR (400 MHz, DMSO-d 6 ) d11.00 (s, 1H), 9.95 (s, 1H), 7.84 (d, J =4.0 Hz, 1H), 7.77 (d, J = 36.0 Hz, 2H), 7.38 (dd, J = 12.0, 8.0 Hz, 1H), 7.13(dd, J = 8.0, 4.0 Hz, 1H), 5.28 (s, 2H), 5.17 – 5.03 (dd, J= 16.0,8.0 Hz,1H),4.40 (dd, J = 60.0, 16.0 Hz, 2H), 3.61 (t, J = 8.0 Hz, 2H), 3.12 (t, J = 8.0Hz, 2H), 2.95 – 2.82 (m, 1H), 2.59 (d, J =20.0 Hz, 1H), 2.25 (s, 3H), 2.00(dd, J = 8.0, 4.0 Hz, 1H), 1.90 (s, 1H).

[0290] MS-ESI: 589.4 [M+H] + . Example 8

[0291] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopentylethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 8)

[0292]

[0293] Step 1: Preparation of ethynylcyclopentane

[0294]

[0295] Ethynylcyclobutane was synthesized from ethynylcyclopentane, except that cyclobutanal was replaced with cyclopentanal. 1.43 g of a brown oil was obtained, with a yield of 30%.

[0296] Step 2: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopentylethynylisoindole-5-carboxylate

[0297]

[0298] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopentylethynylisoindole-5-carboxylic acid methyl ester was the same as (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester), except that trimethylsilyl acetylene was replaced by ethynylcyclopentane. 703 mg of white solid was obtained with a yield of 29%.

[0299] 1 H NMR (400 MHz, CDCl 3 ) d 8.43 (d, J = 4.0 Hz, 1H), 8.25 (d, J = 4.0Hz, 1H), 8.11 (s, 1H), 5.25 (dd, J = 16.0, 8.0 Hz, 1H), 4.45 (dd, J = 64.0,16.0 Hz, 2H), 3.95 (s, 3H), 3.01–2.79 (m, 3H), 2.40 (qd, J = 16.0, 4.0 Hz,1H), 2.25 (ddd, J = 12.0, 6.0, 4.0 Hz, 1H), 2.02 (dd, J = 12.0, 4.0 Hz, 2H),1.81–1.63 (m, 6H).

[0300] Step 3: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopentylethynylisoindole

[0301]

[0302] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopentethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopentethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclopentylethynylisoindole-5-carboxylic acid methyl ester). 133 mg of yellow solid was obtained with a yield of 21%.

[0303] MS-ESI: 367.2 [M+H] + .

[0304] Step 4: Preparation of Compound 8

[0305]

[0306] Compound 8 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclopentethynylisoindole). 6 mg of the target product was obtained in a yield of 13%.

[0307] 1 H NMR (400 MHz, CDCl 3 ) d 8.02 (s, 1H), 7.83 (s, 1H), 7.60 (s, 1H),7.07 (dd, J = 20.0, 12.0 Hz, 2H), 5.29 (d, J = 8.0 Hz, 2H), 4.41 (dd, J =60.0, 16.0 Hz, 2H), 2.98–2.78 (m, 3H), 2.40 (dt, J = 8.0, 4.0 Hz, 1H), 2.27–2.15 (m, 1H), 2.10–1.93 (m, 3H), 1.38–1.20 (m, 8H).

[0308] MS-ESI: 588.4 [M+H] + . Example 9

[0309] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclohexylethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 9)

[0310]

[0311] Step 1: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclohexylethynylisoindole-5-carboxylate

[0312]

[0313] The synthesis of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclohexylethynylisoindole-5-carboxylate) was the same as that of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate) except that trimethylsilyl acetylene was replaced with ethynylcyclohexane. 2.4 g of a white solid was obtained with a yield of 38%.

[0314] 1 H NMR (400 MHz, CDCl 3 ) d 8.42 (d, J = 1.4 Hz, 1H), 8.25 (d, J = 1.4Hz, 1H), 5.25 (dd, J = 13.2, 4.9 Hz, 1H), 4.44 (dd, J = 63.1, 17.3 Hz, 2H), 3.94 (s, 3H), 3.48 (d, J = 4.2 Hz, 1H), 2.88 (m, 3H), 2.63 (m, 1H), 2.40 (qd, J = 13.1, 5.1 Hz, 1H), 2.23 (dtd, J = 13.0, 5.2, 2.6 Hz, 1H), 1.89 (m, 2H), 1.75 (dd, J = 9.3, 3.9 Hz, 2H), 1.53 (dd, J = 9.7, 6.4 Hz, 3H), 1.39 (m, 3H).

[0315] Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclohexylethynylisoindole

[0316]

[0317] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclohexethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclohexethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-cyclohexylethynylisoindole-5-carboxylic acid methyl ester). 320 mg of yellow solid was obtained with a yield of 14%.

[0318] 1 H NMR (400 MHz, CDCl 3 ) d 8.35 (d, J = 1.9 Hz, 1H), 8.21 (d, J = 8.2Hz, 1H), 5.19 (dd, J = 13.1, 5.4 Hz, 1H), 4.75 (m, 2H), 4.54 (s, 1H), 4.44(dd, J = 17.5, 7.2 Hz, 2H), 3.94 (s, 2H), 3.93 (s, 2H), 2.85 (m, 1H), 2.62(dd, J = 22.1, 13.6 Hz, 5H), 2.40 (m, 2H), 2.22 (dd, J = 15.0, 6.9 Hz, 1H),2.07 (dd, J = 24.1, 9.7 Hz, 3H), 1.26 (m, 10H).

[0319] Step 3: Preparation of Compound 9

[0320]

[0321] Compound 9 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-cyclohexethynylisoindole). 21 mg of the target product was obtained in a yield of 14%.

[0322] 1 H NMR (400 MHz, CDCl 3 ) d8.41 (dd, J = 4.3, 2.9 Hz, 1H), 8.23 ​​(dd, J = 3.5, 1.4 Hz, 1H), 7.83 (d, J = 1.5 Hz, 1H), 7.61 (d, J = 1.5 Hz, 1H), 7.09(m, 1H), 7.06 (s, 1H), 6.86 (m, 2H), 5.28 (s, 2H), 5.23 (dd, J = 13.3, 5.2Hz, 2H), 4.40 (dd, J = 59.3, 16.7 Hz, 4H), 2.82 (m, 1H), 2.63 (dd, J = 11.0,7.0 Hz, 2H), 2.39 (m, 2H), 2.23 (m, 2H), 1.37 (m, 11H).

[0323] MS-ESI: 602.4 [M+H] + . Example 10

[0324] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-piperidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 10)

[0325]

[0326] Step 1: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindole-5-carboxylate

[0327]

[0328] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindole-5-carboxylic acid methyl ester was similar to (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester) except that trimethylsilyl acetylene was replaced by 1-Boc-4-ethynylpiperazine. 4.4 g of the target product was obtained in a yield of 81%.

[0329] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.02 (s, 1H), 8.14 (m, 2H), 5.15 (dd, J =13.3, 5.1 Hz, 1H), 4.48 (dd, J = 54.6, 18.7 Hz, 2H), 3.90 (s, 3H), 3.65 (m,2H), 3.17 (m, 2H), 2.94 (m, 2H), 2.60 (d, J = 17.9 Hz, 1H), 2.50 (m, 5H), 2.01 (dd, J = 12.0, 6.6 Hz, 1H), 1.85 (m, 2H), 1.57 (m, 2H), 1.40 (s, 9H).

[0330] Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(4-Boc-piperidinyl)ethynylisoindole

[0331]

[0332] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(4-Boc-piperidinyl)ethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-(4-Boc-piperidinyl)ethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindole-5-carboxylic acid methyl ester. 700 mg of the target product was obtained with a yield of 17%.

[0333] 1 H NMR (400 MHz, CDCl 3 ) d 8.40 (s, 1H), 8.22 (d, J = 12.2 Hz, 1H),7.80 (s, 1H), 7.62 (s, 1H), 5.21 (dd, J = 13.4, 5.1 Hz, 1H), 4.77 (s, 2H), 4.39 (dd, J = 56.4, 16.6 Hz, 3H), 3.94 (s, 3H), 3.74 (d, J= 11.2 Hz, 3H), 3.23 (s, 3H), 2.83 (s, 4H), 2.59 (dd, J = 18.3, 7.0 Hz, 1H), 2.41 (qd, J =13.0, 4.8 Hz, 1H), 2.22 (m, 2H), 2.04 (m, 3H), 1.87 (m, 4H), 1.46 (s, 9H).

[0334] Step 3: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate

[0335]

[0336] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindole was similar to that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole) (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindole) (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole) (40 mg of the target product was obtained in a 27% yield.

[0337] 1 H NMR (400 MHz, CDCl 3 ) d 8.09 (s, 1H), 8.00 (s, 1H), 7.86 (s, 1H), 7.62 (s, 1H), 7.09 (m, 1H), 7.03 (s, 1H), 6.88 (m, 1H), 5.28 (s, 2H), 5.24(dd, J = 13.4, 5.1 Hz, 1H), 4.40 (dd, J = 61.4, 16.7 Hz, 2H), 3.76 (d, J =13.6 Hz, 2H), 3.22 (m, 2H), 2.93 (t, J= 15.3 Hz, 3H), 2.84 (d, J = 5.6 Hz,0H), 2.41 (qd, J = 13.2, 4.7 Hz, 1H), 2.24 (dd, J = 8.1, 4.9 Hz, 1H), 2.01(dd, J = 12.5, 6.8 Hz, 1H), 1.88 (m, 2H), 1.67 (d, J = 9.4 Hz, 3H), 1.47 (s,9H).

[0338] Step 4: Preparation of Compound 10

[0339]

[0340] The synthesis of compound 10 was similar to that of compound 6, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(3-Boc-azetidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate was replaced with (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-Boc-piperidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate. The resulting product was a white solid (52 mg, 97% yield).

[0341] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.00 (s, 1H), 9.93 (s, 1H), 8.71 (s, 1H), 7.84 (s, 2H), 7.74 (s, 1H), 7.39 (m, 1H), 7.14 (d, J = 9.0 Hz, 1H), 5.29 (s,2H), 5.16 (dd, J = 13.3, 5.1 Hz, 1H), 4.41 (dd, J = 60.6, 17.8 Hz, 2H), 3.23(d, J = 12.1 Hz, 2H), 3.07 (dd, J = 12.3, 8.5 Hz, 3H), 2.93 (m, 1H), 2.63 (t, J= 16.2 Hz, 1H), 2.44 (m, 1H), 2.07 (m, 2H), 2.01 (m, 1H), 1.84 (dd, J =11.7, 7.1 Hz, 2H).

[0342] MS-ESI: 603.4 [M+H] + . Example 11

[0343] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-(4-methylpiperidinyl)ethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 11)

[0344]

[0345]

[0346] The synthesis of compound 11 was similar to compound 7, except that compound 6 was replaced by compound 10. 30 mg of a white solid was obtained, with a yield of 74%.

[0347] 1 H NMR (400 MHz, CD 3 OD ) d 7.99 (s, 1H), 7.88 (s, 1H), 7.77 (s, 1H),7.23 (m, 1H), 6.99 (d, J = 9.0 Hz, 1H), 5.33 (s, 2H), 5.19 (dd, J = 13.3, 5.2Hz, 1H), 4.53 (dd, J = 28.7, 17.6 Hz, 3H), 3.41 (s, 2H), 2.93 (m, 1H), 2.86(s, 3H), 2.81 (dd, J = 10.0, 7.5 Hz, 1H), 2.53 (dd, J = 13.1, 4.7 Hz, 1H),2.20 (dd, J = 9.7, 5.0 Hz, 3H), 2.04 (d, J = 4.3 Hz, 2H).

[0348] MS-ESI: 615.3 [MH] -. Example 12

[0349] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-phenylethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 12)

[0350]

[0351] Step 1: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-phenylethynylisoindole-5-carboxylate

[0352]

[0353] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-phenylethynylisoindole-5-carboxylic acid methyl ester was the same as (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester) except that trimethylsilylethynyl acetylene was replaced by phenylacetylene. 2.9 g of white solid was obtained with a yield of 90%.

[0354] 1 H NMR (400 MHz, DMSO-d 6 ) d 11.05 (s, 1H), 8.29 (d, J = 1.4 Hz, 1H),8.20 (d, J = 1.4 Hz, 1H), 7.67 (m, 2H), 7.48 (m, 3H), 5.20 (dd, J = 13.2, 5.1Hz, 1H), 4.62 (dd, J = 60.6, 18.8 Hz, 2H), 3.93 (s, 3H), 2.95 (m, 1H), 2.57(m, 2H), 2.05 (dd, J = 9.6, 4.3 Hz, 1H).

[0355] Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-phenylethynylisoindole

[0356]

[0357] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-phenylethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-phenylethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-phenylethynylisoindole-5-carboxylic acid methyl ester). 1.0 g of yellow solid was obtained with a yield of 37%.

[0358] 1 H NMR (400 MHz, CDCl 3 ) d 8.58 (s, 1H), 8.40 (d, J = 1.4 Hz, 1H), 8.33(d, J = 1.4 Hz, 1H), 7.80 (d, J = 1.5 Hz, 1H), 7.69 (d, J = 1.5 Hz, 1H), 7.52(ddd, J = 7.7, 4.6, 2.6 Hz, 5H), 7.36 (m, 8H), 5.19 (dd, J = 13.3, 5.1 Hz,1H), 5.09 (d, J = 3.0 Hz, 1H), 4.76 (s, 3H), 4.60 (d, J = 7.1 Hz, 1H), 4.45(m, 3H), 3.94 (s, 3H), 2.82 (m, 3H), 2.59 (m, 1H), 2.34 (td, J = 12.9, 5.4Hz, 1H), 2.21 (m, 1H), 2.03 (dd, J = 17.6, 6.8 Hz, 5H).

[0359] Step 3: Preparation of Compound 12

[0360]

[0361] Compound 12 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-phenylethynylisoindole). 16 mg of the target product was obtained in a yield of 10%.

[0362] 1 H NMR (400 MHz, CDCl 3 ) d 8.11 (s, 1H), 8.08 (s, 1H), 7.90 (d, J = 1.5Hz, 1H), 7.75 (d, J = 1.5 Hz, 1H), 7.54 (m, 3H), 7.38 (m, 4H), 7.09 (m, 3H), 6.87 (m, 1H), 5.32 (s, 3H), 5.26 (dd, J = 13.3, 5.2 Hz, 1H), 4.53 (dd, J =60.9, 16.8 Hz, 3H), 2.87 (m, 3H), 2.41 (qd, J = 13.1, 5.0 Hz, 1H), 2.24 (m,2H).

[0363] MS-ESI: 594.3 [MH] - . Example 13

[0364] Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-tert-butylethynylisoindol-5-yl)methyl(2-fluoro-5-trifluoromethoxy)phenyl)carbamate (Compound 13)

[0365]

[0366] Step 1: Preparation of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-tert-butylethynylisoindole-5-carboxylate

[0367]

[0368] The synthesis of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-tert-butylethynylisoindole-5-carboxylate) was the same as that of methyl (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylate) except that trimethylsilylethynyl acetylene was replaced with tert-butylacetylene. 1.8 g of a white solid was obtained with a yield of 59%.

[0369] 1 H NMR (400 MHz, CDCl 3 ) d 8.44 (s, 1H), 8.42 (d, J = 1.4 Hz, 1H), 8.24(d, J = 1.5 Hz, 1H), 5.26 (dd, J = 13.4, 5.1 Hz, 2H), 4.43 (dd, J = 64.1,17.3 Hz, 3H), 3.94 (s, 4H), 2.88 (m, 3H), 2.40 (qd, J = 13.0, 5.3 Hz, 1H), 2.23 (m, 1H), 1.32 (s, 9H).

[0370] Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-tert-butylethynylisoindole

[0371]

[0372] The synthesis of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-tert-butylethynylisoindole was the same as that of (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-tert-butylethynylisoindole, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-trimethylsilylethynylisoindole-5-carboxylic acid methyl ester was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-7-tert-butylethynylisoindole-5-carboxylic acid methyl ester). 28 mg of yellow solid was obtained with a yield of 11%.

[0373] 1 H NMR (400 MHz, CDCl 3 ) d 7.71 (d, J = 1.6 Hz, 1H), 7.55 (d, J= 1.5Hz, 1H), 5.00 (dd, J = 12.8, 5.9 Hz, 1H), 4.72 (s, 2H), 4.33 (q, J = 17.1 Hz,2H), 2.52 (m, 1H), 2.39 (ddd, J = 26.3, 13.2, 2.9 Hz, 1H), 2.21 (d, J = 14.6Hz, 1H), 2.08 (m, 1H), 1.99 (m, 1H), 1.33 (s, 9H).

[0374] Step 3: Preparation of Compound 13

[0375]

[0376] Compound 13 was synthesized in the same manner as compound 1, except that (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-ethynylisoindole was replaced by (2-(2,6-dioxopiperidin-3-yl)-3-oxo-5-hydroxymethyl-7-tert-butylethynylisoindole). 5 mg of the target product was obtained in a yield of 12%.

[0377] 1 H NMR (400 MHz, CDCl 3 ) d 8.10 (s, 1H), 7.81 (s, 1H), 7.57 (s, 1H),7.07 (m, 2H), 6.87 (dd, J = 8.6, 4.2 Hz, 1H), 6.66 (s, 1H), 5.26 (s, 2H), 5.06 (dd, J = 12.8, 5.8 Hz, 1H), 4.59 (m, 1H), 4.37 (dd, J = 49.9, 17.2 Hz,2H), 2.39 (qd, J = 13.0, 3.0 Hz, 1H), 2.21 (m, 1H), 2.10 (dt, J = 14.0, 3.3Hz, 1H), 1.98 (m, 1H), 1.34 (s, 9H).

[0378] MS-ESI: 576.2 [M+H] + .

[0379] Test Example 1

[0380] Western Blot analysis of GSPT1 protein expression in cells after compound treatment

[0381] After KG-1 cells were digested and counted, the cell suspension was evenly added to the corresponding wells in a 15-well plate at an appropriate cell volume; after shaking, the cells were cultured in a cell culture incubator, and the drug was administered after 12 hours of cell attachment.

[0382] Dosage groups: DMSO alone group, 1nM, 10nM, 100nM, samples were collected 4 hours after administration.

[0383] Western Blot

[0384] 1) Pretreatment: Rinse cells three times with pre-chilled PBS. Add an appropriate amount of pre-chilled lysis buffer, harvest with a cell scraper, transfer to a centrifuge tube using a pipette, and repeatedly pipette. Lyse on ice for 30 minutes. Collect cells into EP tubes and sonicate (100-200W) for 5 seconds three times. Centrifuge at 12,000 rpm at 4°C for 5 minutes. Transfer the supernatant to a 1.5 ml EP tube. Add an appropriate amount of 6x loading buffer and boil at 95°C for 5 minutes.

[0385] 2) Glue preparation: Select the appropriate concentration of separation gel according to the molecular weight of the target protein

[0386] 3) Sample Loading: Place the gel plate in the electrophoresis tank, add electrophoresis buffer, and slowly remove the comb. Thaw the protein sample from the -80°C freezer at 4°C, then vortex to mix thoroughly. Load 10 μL of sample onto the plates in the order listed.

[0387] 4) Electrophoresis: Connect the electrophoresis device to the power supply and run at 80V constant voltage for about 30 minutes. When the indicator bromofin has entered the separation gel, switch to 120V constant voltage electrophoresis. When the bromofin has reached the bottom of the gel or just exited the gel, turn off the power and remove the gel plate.

[0388] 5) Transfer: Cover the membrane in the order of sponge pad, filter paper, glue, PVDF membrane, filter paper, and sponge pad. Remove any bubbles. Secure the cartridge and place the transfer cassette into the transfer tank, following the corresponding positive and negative electrode configurations. Add pre-chilled transfer solution and place the entire transfer tank in an ice-water mixture (to prevent high temperatures from affecting transfer efficiency). Transfer at 300mA for 1.5 hours.

[0389] 6) Blocking: After transfer, remove the PVDF membrane and mark the front and back of the membrane. Immerse the membrane in 5% skim milk in 1× TBST. Place on a shaker at 70-80 rpm and block at room temperature for 1.5 hours. After blocking, remove the PVDF membrane, add 1× TBST, and wash the membrane three times on a shaker for 5 minutes each.

[0390] 7) Primary Antibody Incubation: Prepare the primary antibody incubation solution by diluting the antibody according to the antibody manufacturer's instructions in the appropriate ratio with antibody diluent. Add the primary antibody to the target protein and incubate overnight at 4°C on a shaker at 70-80 rpm. The next day, recover the primary antibody incubation solution and wash the PVDF membrane three times with 1x TBST for 10 minutes each wash.

[0391] 8) Incubation with secondary antibody: Select a secondary antibody that matches the source of the primary antibody. Dilute the secondary antibody in 5% skim milk or BSA at the appropriate ratio. Place the PVDF membrane in the secondary antibody and incubate on a shaker at 70-80 rpm at room temperature for 1.5 hours. Wash the membrane four times with 1× TBST for 10 minutes each.

[0392] 9) Development: Prepare ECL luminescent solution (mix equal volumes of Solution A and Solution B), drain the liquid from the membrane surface, and evenly drop the luminescent solution onto the protein side of the membrane. Use a gel imaging system to set different exposure times, scan, and analyze the exposure.

[0393] The degradation activity data of the compounds are shown in Table 1.

[0394] Table 1

[0395]

[0396] +++, DC 50 ≤20nM; ++, 20nM <DC 50 ≤200nM; +, DC 50 >200nM.

[0397] It can be seen from the above activity data that some compounds of the present invention have strong degradation activity on GSPT1 protein.

[0398] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations of the claims. Various modifications and variations may be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments may be arbitrarily combined to form additional embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments merely illustrate several implementations of the present invention and do not limit the scope of protection of the patent of this invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound is any one of the following: 、 、 、 、 、 、 、 、 、 、 。 2. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound is any one of the following: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 3. A pharmaceutical composition comprising the compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof.

4. Use of the compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 3, in the preparation of a medicament for preventing and / or treating a GSPT1-related disease.

5. The use according to claim 4, characterized in that The GSPT1-related disease is selected from cancer, autoimmune disease, immunodeficiency disease, aging, viral infection and organ transplant rejection.

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