Compound having dual degradation activity against IRAK4 and FLT3 proteins, pharmaceutical composition comprising same, and use thereof

Abstract

The present disclosure belongs to the field of pharmaceutical chemistry, and in particular relates to a compound having a dual degradation activity against IRAK4 and FLT3 proteins, a pharmaceutical composition comprising same, and the use thereof. Provided in the present disclosure is a compound having a structure of formula (I), which can effectively degrade IRAK4 and / or FLT3 proteins in cells, and can be used for preventing and / or treating diseases and / or conditions that are at least partially responsive to IRAK4 and / or FLT3.

Description

Compound with dual degradation activity of IRAK4 and FLT3 proteins, pharmaceutical composition containing same, and use thereof

[0001] Citation of Related Applications

[0002] This disclosure claims priority to an invention patent application filed with the Patent Office of China on December 20, 2023, with application number 202311765223.2 and titled “Compounds having dual degradation activity of IRAK4 and FLT3 proteins, pharmaceutical compositions containing the same, and applications thereof,” and the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present disclosure belongs to the field of medical technology and relates to a compound having dual degradation activity of IRAK4 and FLT3 proteins, a pharmaceutical composition containing the same, and its use in medicine, in particular, its use in preventing and / or treating related diseases or conditions (including tumors, inflammatory diseases, and autoimmune diseases) that are at least partially responsive to IRAK4 and / or FLT3 mediation. Background Art

[0004] Interleukin-1 receptor-associated kinase 4 (IRAK4) is a serine / threonine protein kinase that plays a crucial role in the innate immune response and is a key signaling factor in downstream inflammatory signaling pathways mediated by the interleukin receptor family (IL-1, IL-18, IL-33) and the Toll-like receptor family (excluding TLR3) (Clin. Immunol. 2010, 135, 210-222). Binding of extracellular signaling molecules to interleukin receptors or Toll-like receptors recruits the intracellular Myddosome, a multiprotein complex consisting of MyD88, IRAK4, and IRAK1 / 2 (Nature 2010, 465, 885-890). This complex then phosphorylates and activates IRAK1 / 2 and the downstream NF-κB and MAPK signaling pathways, promoting the release of related proinflammatory cytokines (Mol. Cell 2014, 55, 891-903). Among them, IRAK4, as a key regulatory protein, can control TLR / IL-1R-mediated inflammatory signals through its kinase and scaffold functions (Biochem Pharmacol 2010, 80, 1981-1991).

[0005] When the IRAK4-mediated signaling pathway is overactivated, the body will experience a surge in inflammatory factors, which is closely related to a variety of autoimmune diseases, such as rheumatoid arthritis (Mol.Med.2010,16,552-557), systemic lupus erythematosus (PNAS 2009,106,12061-12066), inflammatory bowel disease (J.Exp.Med.2012,209,1595-1609), psoriasis (J.Invest.Dermatol 2011,131,2428-2437) and airway inflammation (J.Allergy Clin.Immunol 2012,130,1159-1166). In addition, high levels of IRAK4 phosphorylation have been found in a variety of tumors, such as hematopoietic malignancies (Curr. Opin. Hematol 2022, 29, 8-19), colon cancer (JCI insight 2019, 4, e130867), and melanoma (Cancer Res 2012, 72, 6209-6216). Studies have found that mutations in the splicing factor U2AF1 / SF3B1 gene can also cause intracellular protein miscleavage, leading to overexpression of the oncogenic IRAK4-L (IRAK4 Long Form) isoform, abnormally and persistently activating the downstream NF-κB signaling pathway, and are associated with the occurrence and poor prognosis of myelodysplastic syndrome (MDS) / acute myeloid leukemia (AML) (Nat. Cell Biol 2019, 21, 640-650). Animal experiments have shown that knocking out or inhibiting IRAK4 can prevent the release of inflammatory factors mediated by IL-1R or TLR (Nature 2002, 416, 750-756) or inhibit the progression of related tumors (J. Exp. Med 2015, 212, 2189-2201). Another study showed that patients with congenital IRAK4 protein deficiency are not lethal and only have a certain degree of infection risk (such as Streptococcus pneumoniae), and the infection rate decreases with age (J. Pediatr 2004, 144, 524-526). In summary, IRAK4 has become a safe and attractive target for the treatment of inflammatory diseases and malignancies.

[0006] Although the IRAK4-mediated signal transduction pathway is mainly related to its kinase function, there are also reports that in some cell types (human skin fibroblasts), the IRAK4 kinase function is redundant, and its protein scaffold function is more important for IL-1β signal transduction. At present, there are several IRAK4 small molecule inhibitors entering clinical research, such as PF-06650833 developed by Pfizer (J.Med.Chem.2017,60,5521-5542), which is currently in Phase II clinical trials for the treatment of rheumatoid arthritis and hidradenitis suppurativa; Bayer has successively launched two IRAK4 inhibitors (BAY-1830839 and BAY-1834845), which are currently in Phase I clinical trials; Rigel's R-835 is in Phase I clinical trials; and CA4948 developed by Curis is mainly used for AML / MDS patients (ACS Med.Chem.Lett.2020,11,2374-2381). However, these small molecule inhibitors can only inhibit the kinase function of IRAK4, and traditional small molecule inhibitors are prone to drug resistance. Therefore, inhibiting IRAK4 kinase activity alone may not be sufficient to produce optimal therapeutic effects.

[0007] FMS-like tyrosine kinase 3 (FLT3) belongs to the Type III receptor tyrosine kinase family. It is mainly expressed in a variety of lymphoid hematopoietic stem cells and tissues and is involved in the proliferation and differentiation of hematopoietic stem cells (J.Med.Chem 2021, 64, 2878-2900). FLT3 is a transmembrane protein composed of an extracellular receptor region, a transmembrane region, a juxta-membrane region, and an intracellular kinase structure (Mol.Cell 2004, 13, 169-178). When bound to the ligand outside the cell, FLT3 dimerizes and self-phosphorylates, activating downstream signal transduction, including STAT5A, RAS / MAPK, PI3K / AKT and other pathways, promoting the survival, growth and differentiation of bone marrow hematopoietic stem cells (J.Med.Chem 2018, 61, 3855-3869). When the FLT3 signaling pathway is overactivated, it will lead to malignant clonal diseases of hematopoietic stem cells. The affected cells (leukemia cells) will experience uncontrolled proliferation, differentiation disorders, and blocked apoptosis, which will eventually induce the clinical manifestations of AML. FLT3 mutations are closely related to the progression and prognosis of AML. About 30% of newly diagnosed AML patients carry different FLT3 mutation phenotypes (Ann.Hematol 2017,96,1993-2003). Internal tandem duplication mutations (FLT3-ITD) in the juxtamembrane region account for about 25% of AML patients. This mutation weakens the kinase autoinhibition function of the juxtamembrane region through conformational changes, causing FLT3 kinase to be in a state of continuous activation (Cancer Science 2020,111,312-322). Point mutations in the FLT3 tyrosine kinase domain (FLT3-TKD) mainly involve D835, F691, N676, Y842, etc., accounting for about 10% of AML patients. FLT3-TKD also leads to non-ligand-activated dimerization and self-phosphorylation, which keeps the FLT3 downstream signaling pathway in a state of continuous activation and may lead to resistance to some kinase inhibitors (Cancers 2022, 14, 3398).

[0008] Currently, the main clinical treatments for AML are chemotherapy and stem cell transplantation. First-generation FLT3 inhibitors, including tandutinib, sunitinib, midostaurin, lestaurtinib, and sorafenib, are primarily non-selective kinase inhibitors. Their monotherapy efficacy in AML is poor and has a high incidence of off-target adverse reactions. Second-generation FLT3 inhibitors, including gilteritinib, quizartinib, and crenolanib, have higher selectivity and stronger inhibitory activity (Leukemia 2019, 33, 299-312). However, long-term use of FLT3 kinase inhibitors may lead to clinical drug resistance, resulting in a decrease in the efficacy of FLT3 small molecule inhibitors and an increase in patient relapse rates. Analysis of samples from AML patients who relapsed after midostaurin treatment identified N676K as the primary cause of drug resistance. In patients with AML resistance after quizartinib treatment, multiple point mutations were found in the activation loop of the FLT3 kinase domain, including D835Y, D835V, D835F and gating residue F691L mutations, which induced acquired resistance mutations in tumor cells (Int.J.Mol.Sci.2020,21,1537). Studies have found that in the later stages of AML treatment with targeted FLT3 inhibitors, patients will develop acquired resistance due to bypass activation. The resistance mechanism may be related to innate immune stress activated by the IRAK4 / IRAK1 complex, which compensatorily activates cell growth-promoting or survival-promoting signaling pathways (Sci.Transl.Med.2019,11,8828). Therefore, simultaneously targeting the FLT3 and IRAK4 signaling pathways has potential clinical application value.

[0009] Protein degradation targeting chimera (PROTAC) is a technology different from traditional small molecule inhibitors. It is a special heterogeneous bifunctional molecule. One end is a small molecule inhibitor that can bind to the target protein, and the other end is a ligand that can recognize E3 ubiquitin ligase, connected in the middle by a connecting chain. PROTAC molecules can bring the target protein and E3 ubiquitin ligase closer to form a ternary complex, and ultimately degrade the target protein through the ubiquitin-proteasome pathway. PROTAC technology uses an "event-driven" mechanism of action to degrade the content of the target protein in the cell in a trace and efficient manner, avoiding long-term, high-dose administration that leads to increased target protein expression, target protein mutations, and drug resistance caused by bypass signal activation, thereby producing better and wider therapeutic effects (Nature Biotechnology 2020, 38, 1221-1223).

[0010] Currently, PROTAC technology has been used in the development of drugs for a variety of targets, such as androgen receptors, estrogen receptors, and BTK. In recent years, pharmaceutical companies such as Nurix (WO 2023023255), C4 (WO 2023283372), Arvinas (WO 2022266258), Kymera (WO2020264499), and domestic companies such as Lingtai (WO 2023025159) and Hisense (CN 114437035) are developing IRAK4 protein degradation technology. FLT3 protein degraders (WO 202225328; CN 116444495; CN 115124590) have also been studied. However, research on degraders that can degrade both IRAK4 and FLT3 proteins has not yet been disclosed. Therefore, developing novel degraders with dual degradation activity of IRAK4 and FLT3 proteins, while inhibiting the signaling pathways mediated by FLT3 and IRAK4, has the potential to enhance the clinical therapeutic effect of FLT3-mutated AML patients and improve the drug resistance problem of clinical small molecule therapy. Summary of the Invention

[0011] Problems to be solved by the invention

[0012] The purpose of the present disclosure is to provide a compound having dual degradation activity of IRAK4 and FLT3 proteins, a pharmaceutical composition containing the same, and applications thereof.

[0013] Solutions for solving problems

[0014] The purpose of this disclosure is achieved through the following technical solutions:

[0015] <First Aspect>

[0016] The present disclosure provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt or solvate thereof:

[0017] in,

[0018] R 1 Select one of the following fragments:

[0019] ULM is selected from one of the following segments:

[0020] L m is a linker fragment, which consists of m identical, partially identical or different L groups and connects the ULM and the piperazine ring through a covalent bond;

[0021] m is selected from 1, 2, 3, 4 and 5;

[0022] Each L is independently selected from the following groups:

[0023] 1) having 1 to 3 independently selected R b Substituted C 3-12 cycloalkylene;

[0024] 2) having 1 to 3 independently selected b Substituted C 6-10 arylene;

[0025] 3) having 1 to 3 independently selected R b substituted 4- to 12-membered heterocyclylene;

[0026] 4) having 1 to 3 independently selected b substituted 5- to 12-membered heteroarylene;

[0027] 5) having 1 to 3 independently selected R d Substituted C 1-12 alkylene;

[0028] 6) having 1 to 3 independently selected d Substituted C 2-12 vinylidene;

[0029] 7) having 1 to 3 independently selected d Substituted C 2-12 ethynylene;

[0030] 8) having 1 to 6 ethylene glycol or propylene glycol units; and

[0031] 9)-C(O)-, -C(O)O-, -O-, -N(R c )-, -S-, -S(O)-, -C(S)-, -C(S)O-, -S(O)2-, -S(O)N(R c )-、-S(O)2N(R c )-、-C(O)-N(R c )-、-N(R c )C(O)-N(R c )-and-OC(O)-N(R c )-;

[0032] If present, R 4 Selected from hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, C 5-12 Heteroaryl, -OR c 、-C(O)R c 、-C(O)OR c 、-C(O)-N(Rc )2、-N(R c )2、-N(R c )C(O)-R c 、-N(R c )C(O)OR c 、-N(R c )C(O)N(R c )2、-OC(O)R c 、-OC(O)-N(R c )2、-SR c 、-S(O)R c 、-S(O)2R c and -S(O)2-N(R c )2;

[0033] If present, each R b are each independently selected from hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, C 5-12 Heteroaryl, -OR c 、-C(O)R c 、-C(O)OR c 、-C(O)-N(R c )2、-N(R c )2、-N(R c )C(O)-R c 、-N(R c )C(O)OR c 、-N(R c )C(O)N(R c )2、-OC(O)R c 、-OC(O)-N(R c )2、-SR c 、-S(O)R c 、-S(O)2R c and -S(O)2-N(R c )2; if present, the C 1-6 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, C 5-12 The heteroaryl group may be substituted with 1 to 3 R d Replacement; if present, any two non-adjacent R b A bridged ring can be formed through carbon atoms; or two R b Together with the carbon atom to which it is attached, it forms a spiro ring;

[0034] If present, each R care each independently selected from hydrogen, C 1-6 Alkyl and C 3-6 Cycloalkyl;

[0035] If present, each R d Each is independently selected from hydrogen, deuterium, halogen, hydroxy, amino, methylamino, dimethylamino, cyano, methyl, deuterated methyl, methoxy and deuterated methoxy.

[0036] In one embodiment of the present disclosure, L m is a linker fragment, which consists of m identical, partially identical or different L groups and connects the ULM and the piperazine ring through a covalent bond;

[0037] m is selected from 1, 2, 3, 4 and 5;

[0038] Each L is independently selected from the following groups:

[0039] 1) having 1 to 3 independently selected R b Substituted C 3-12 cycloalkylene;

[0040] 2) having 1 to 3 independently selected b Substituted C 6-10 arylene;

[0041] 3) having 1 to 3 independently selected R b substituted 4- to 12-membered heterocyclylene;

[0042] 4) having 1 to 3 independently selected b substituted 5- to 12-membered heteroarylene;

[0043] 5) having 1 to 6 ethylene glycol or propylene glycol units; and

[0044] 6)-C(O)-, -C(O)O-, -O-, -N(R c )-、-C(O)-N(R c )-、-N(R c )C(O)-N(R c )-and-OC(O)-N(R c )-;

[0045] The R b and R c As defined in claim 1.

[0046] In one embodiment of the present disclosure, L m is a linker fragment, which consists of m identical, partially identical or different L groups and connects the ULM and the piperazine ring through a covalent bond;

[0047] m is selected from 1, 2, 3, 4 and 5;

[0048] Each L is independently selected from the following groups:

[0049] (1)-C(O)-, -CH2-, -C(D)(H)-, -C(D)2-, -CR c F-、-CF2-、-O-、-N(R c )-、-C(O)-N(R c )-、

[0050] -N(R c )C(O)-N(R c )-and-OC(O)-N(R c )-;and

[0051] (2) One of the following fragments:

[0052] The R c As defined in claim 1.

[0053] In a preferred embodiment of the present disclosure, at least one L is selected from one of the following fragments:

[0054] In one embodiment of the present disclosure, L m Select one of the following fragments:

[0055] In one embodiment of the present disclosure, L m Select one of the following fragments:

[0056] <Second Aspect>

[0057] The present disclosure provides the following compounds or pharmaceutically acceptable salts or solvates thereof:

[0058] <Third Aspect>

[0059] The present disclosure provides a pharmaceutical composition comprising the compound according to <the first aspect> or <the second aspect> or a pharmaceutically acceptable salt or solvate thereof.

[0060] Furthermore, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.

[0061] <Fourth Aspect>

[0062] The present disclosure provides a pharmaceutical combination comprising the compound according to the first aspect or the second aspect, or a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition according to the third aspect.

[0063] Furthermore, the pharmaceutical combination further comprises at least one additional drug for preventing and / or treating diseases and / or conditions responsive to IRAK4 and / or FLT3.

[0064] <Fifth Aspect>

[0065] The present disclosure provides the use of the compound according to <First Aspect> or <Second Aspect> or its pharmaceutically acceptable salt or solvate, or the pharmaceutical composition according to <Third Aspect>, or the drug combination according to <Fourth Aspect> in the preparation of a medicament for preventing and / or treating diseases and / or conditions that are at least partially responsive to IRAK4 and / or FLT3.

[0066] Furthermore, the diseases and / or conditions include: lymphoma, leukemia, myelodysplastic syndrome, endometriosis, atopic dermatitis, hidradenitis suppurativa, psoriasis, systemic lupus erythematosus, multiple sclerosis or rheumatoid arthritis.

[0067] <Sixth Aspect>

[0068] The present disclosure provides a compound according to the <First Aspect> or <Second Aspect> or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to the <Third Aspect>, or a drug combination according to the <Fourth Aspect>, for preventing and / or treating diseases and / or conditions that are at least partially responsive to IRAK4 and / or FLT3.

[0069] Furthermore, the diseases and / or conditions include: lymphoma, leukemia, myelodysplastic syndrome, endometriosis, atopic dermatitis, hidradenitis suppurativa, psoriasis, systemic lupus erythematosus, multiple sclerosis or rheumatoid arthritis.

[0070] <Seventh Aspect>

[0071] The present disclosure provides a method for preventing and / or treating diseases and / or conditions that are at least partially responsive to IRAK4 and / or FLT3, comprising the following steps: administering a preventively and / or therapeutically effective amount of a compound according to <the first aspect> or <the second aspect> or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to <the third aspect>, or a drug combination according to <the fourth aspect> to a patient in need thereof.

[0072] Furthermore, the diseases and / or conditions include: lymphoma, leukemia, myelodysplastic syndrome, endometriosis, atopic dermatitis, hidradenitis suppurativa, psoriasis, systemic lupus erythematosus, multiple sclerosis or rheumatoid arthritis.

[0073] Effects of the present invention

[0074] The compounds disclosed herein having dual IRAK4 and FLT3 protein degradation activities can be used to prevent and / or treat diseases and / or conditions that are at least partially responsive to IRAK4 and / or FLT3. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] FIG1 is a western blot test result of IRAK4 protein degradation by compounds A3, A4, A5 and A6 in THP-1 cells.

[0076] FIG2 is the western test result of compound A3, A6, and A8 on FLT3 protein degradation in MOLM-13 cells.

[0077] FIG3 is a graph showing the inhibition of compound A3 on MOLM-13 tumor cell proliferation. DETAILED DESCRIPTION

[0078] In order to have a clearer understanding of the technical features, objectives and beneficial effects of the present disclosure, the technical solution of the present disclosure is now described in detail below, but it should not be understood as limiting the scope of implementation of the present disclosure.

[0079] In order to more clearly describe the content of this disclosure, the terms involved are defined as follows:

[0080] The term "halogen", alone or in combination, denotes fluorine, chlorine, bromine or iodine, in particular fluorine, chlorine or bromine.

[0081] The term "C 1-6 “Alkyl”, alone or in combination, refers to a saturated straight or branched chain alkyl group containing 1 to 6 (particularly 1 to 3) carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, n-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3,-dimethyl-2-butyl. Preferably, “C 1-6 "Alkyl" is any of methyl, ethyl, n-propyl, isopropyl, and tert-butyl. Similarly, the term "C 1-3The term "alkyl" alone or in combination refers to a saturated straight-chain or branched alkyl group containing 1 to 3 carbon atoms, including methyl, ethyl, propyl, isopropyl, and the like.

[0082] The term "OC 1-6 "Alkyl" alone or in combination means "C 1-6 Alkyl-O-" group, where "C 1-6 “Alkyl” is as defined above and includes, but is not limited to, methoxy (—OCH3), ethoxy (—OCH2CH3), n-propoxy (—OCH2CH2CH3), isopropoxy (—OCH(CH3)2), n-butoxy (—OCH2CH2CH2CH3), sec-butoxy (—OCH(CH3)CH2CH3), isobutoxy (—OCH2CH(CH3)2), tert-butoxy (—OC(CH3)3), n-pentoxy (—OCH2CH2CH2CH2CH3) and neopentoxy (—OCH2C(CH3)3), etc.

[0083] The term "C 3-6 "Cycloalkyl" alone or in combination means a saturated or partially unsaturated monocyclic or polycyclic cycloalkyl group containing 3 to 6 (especially 3 to 5) carbon atoms, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0084] The term "3- to 8-membered heterocyclyl" alone or in combination refers to a saturated or partially unsaturated monocyclic heterocyclyl containing 3 to 8 (particularly 3 to 6, more particularly 5 or 6) carbon atoms and heteroatoms or heteroatom groups forming the ring, wherein the heteroatoms or heteroatom groups are selected from O, S, NH, N, P(=O), S(=O) and S(=O)2. "3- to 8-membered heterocyclyl" includes, but is not limited to, aziridinyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxothiomorpholinyl, butyrolactamyl, valerolactamyl, caprolactamyl, butyrolactonyl, valerolactonyl or caprolactonyl, and the like.

[0085] The term "6- to 10-membered bridged heterobicyclic group" alone or in combination refers to a saturated or partially unsaturated bridged heterobicyclic group containing 6 to 10 (particularly 6 to 8) carbon atoms and heteroatoms or heteroatom groups for forming the ring, wherein the heteroatoms or heteroatom groups are selected from O, S, NH, N, P(=O), S(=O) and S(=O)2. "6- to 10-membered bridged heterobicyclic group" includes, but is not limited to, 3,6-diazabicyclo[3.1.1]hept-1-yl, 3,6-diazabicyclo[3.1.1]hept-3-yl, 3,6-diazabicyclo[3.1.1]hept-6-yl and the like.

[0086] The term "5- to 12-membered spiroheterobicyclic radical" alone or in combination denotes a saturated or partially unsaturated bicyclic radical comprising 5 to 12 (in particular 6 to 9) carbon atoms and heteroatoms or heteroatom groups forming the ring, the bicyclic rings being linked to each other via one carbon atom, the heteroatom or heteroatom group being selected from O, S, NH, N, P(=O), S(=O) and S(=O)2. “5- to 12-membered spiroheterobicyclic group” includes, but is not limited to, 4,7-diazaspiro[2.5]oct-7-yl, 1-oxa-6-azaspiro[3.3]hept-6-yl, 2-oxa-6-azaspiro[3.3]hept-6-yl, 2,5-diazaspiro[3.4]oct-2-yl, 5-oxa-2-azaspiro[3.4]oct-2-yl, 6-oxa-2-azaspiro[3.4]oct-2-yl, 1-oxa-7-azaspiro[3.5]non-7-yl, and the like.

[0087] The term "6- to 12-membered heterobicyclic group" alone or in combination refers to a saturated or partially unsaturated bicyclic group containing 6 to 12 (particularly 7 to 10) carbon atoms and heteroatoms or heteroatom groups forming the ring, wherein the bicyclic rings share a carbon-carbon bond, a carbon-hetero bond, or a heterohetero bond, and the heteroatoms or heteroatom groups are selected from O, S, NH, N, P(=O), S(=O), and S(=O)2. "6- to 12-membered heterobicyclic group" includes (but is not limited to) 3,7-diazabicyclo[3.3.0]octan-3-yl, 3,6-diazabicyclo[3.3.0]octan-3-yl, 2,7-diazabicyclo[3.3.0]octan-2-yl, and the like.

[0088] The term "C 6-10 The term "aryl" alone or in combination refers to any stable 6- to 10-membered monocyclic or bicyclic aromatic group, including but not limited to phenyl, naphthyl, tetrahydronaphthyl, 2,3-dihydroindanyl, biphenyl, and the like.

[0089] The term "5- to 10-membered heteroaryl" alone or in combination refers to any stable 5- to 10-membered monocyclic or bicyclic aromatic group, wherein a carbon atom in the ring is replaced by at least one heteroatom or heteroatom group selected from O, S, NH, N, P(=O), S(=O) and S(=O)2, including but not limited to thienyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyridin-2(1H)-onyl, pyridin-4(1H)-onyl, pyrrolyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, imidazolyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, naphthyl, benzothiophenyl, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinolinyl and quinazolinyl.

[0090] The term "7- to 10-membered fused heterobicyclic group" alone or in combination refers to a fused bicyclic group composed of one aryl group and one heteroaryl group or two heteroaryl groups, including (but not limited to) benzothiophenyl, indolyl, benzimidazolyl, pyridoimidazolyl, pyridazinoimidazolyl, pyrazolopyrimidinyl, benzothiazolyl, benzofuranyl, quinolyl, isoquinolyl and quinazolinyl.

[0091] The term "alkenyl" alone or in combination means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond. 2-12 "Alkenyl" refers to a straight or branched chain alkenyl containing 2-12 carbon atoms, including (but not limited to) vinyl, 1-propenyl, 1-butenyl, etc., preferably "C 2-6 "Alkenyl".

[0092] The term "alkynyl" alone or in combination means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon-carbon triple bond. 2-12 "Alkynyl" refers to a straight or branched alkenyl containing 2-12 carbon atoms, including but not limited to ethynyl, 1-propynyl, 1-butynyl, etc., preferably "C 2-6 Alkynyl".

[0093] The term "alkylene" alone or in combination refers to a divalent group, such as alkylene refers to a divalent alkyl group, alkenylene refers to a divalent alkenyl group, alkynylene refers to a divalent alkynyl group, cycloalkylene refers to a divalent cycloalkyl group, heterocycloalkylene refers to a divalent heterocycloalkyl group, arylene refers to a divalent aryl group, and heteroarylene refers to a divalent heteroaryl group. The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, and arylheteroaryl groups are as defined above.

[0094] The term "amino" alone or in combination means a primary amino group (-NH2), a secondary amino group (-NH-) or a tertiary amino group.

[0095] The term "NH(C 1-6 alkyl)", "N(C 1-6 Alkyl)2", "NH(C 3-6 Cycloalkyl)", "N(C 3-6 Cycloalkyl)2" alone or in combination represents an amino group as defined above, wherein the hydrogen atoms in the amino group are replaced by one or two C 1-6 Alkyl or C 3-6 is substituted by a cycloalkyl group, wherein "C 1-6 Alkyl" and "C 3-6 "Cycloalkyl" is as defined above.

[0096] The term "pharmaceutically acceptable salt" means that the compounds of the present disclosure exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts. Pharmaceutically acceptable salts are described in the pharmaceutical salts described by SM Berge in J. Pharmaceutical Sciences (Volume 66: pages 1-19, 1977). In the present disclosure, pharmaceutically acceptable non-toxic acid addition salts refer to salts formed by the compounds of the present disclosure with organic or inorganic acids, including (but not limited to) hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid, and malic acid. Pharmaceutically acceptable non-toxic base addition salts refer to salts of the compounds of the present disclosure formed with organic or inorganic bases, including but not limited to alkali metal salts such as lithium, sodium or potassium salts; alkaline earth metal salts such as calcium or magnesium salts; organic base salts such as ammonium salts or N-containing organic bases. + (C 1-6 alkyl) salt, preferably lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, calcium carbonate, ammonia water, triethylamine or tetrabutylammonium hydroxide, etc. "Pharmaceutically acceptable salts" can be synthesized by general chemical methods.

[0097] The term "solvate" refers to an association formed between one or more solvent molecules and a compound of the present disclosure. Solvents that form solvates include, but are not limited to, water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, and the like.

[0098] The term "each independently" means that at least two groups (or ring systems) present in a structure with the same or similar value ranges may have the same or different meanings in specific circumstances. For example, if substituent X and substituent Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y may be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl. Similarly, when substituent Y is hydrogen, substituent X may be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl.

[0099] The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both occurring and not occurring.

[0100] The term "prevent" refers to completely or nearly completely stopping a disease or condition (e.g., infection, ischemia or reperfusion injury) from occurring, for example, when a patient or subject is susceptible to or at risk for the disease or condition; prevention can also include inhibition, i.e., arresting the development of a condition.

[0101] The terms "treat," ...

[0102] The term "additional" refers to a situation where there is a difference or difference compared to the substance, element or object involved in the present disclosure; for example, one or more additional drugs refer to one or more drugs with a different structure, composition or mechanism of action compared to the substance of the present disclosure.

[0103] The term "effective amount" refers to that amount of an active compound or agent that elicits the biological or medical response in a tissue, system, animal, individual or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.

[0104] Unless otherwise specified, the experimental methods described in the following preparation examples are conventional methods; the reagents and materials described are all commercially available unless otherwise specified.

[0105] In the following preparation examples, all solvents and drugs used were analytically pure or chemically pure; the solvents were redistilled before use; and the anhydrous solvents were treated according to standard methods or literature methods.

[0106] Column chromatography silica gel (100-200 mesh) and thin-layer chromatography silica gel (GF254) were products of Qingdao Ocean Chemical Plant and Yantai Chemical Plant. Unless otherwise specified, petroleum ether (60-90° C.) / ethyl acetate (v / v) was used as the eluent.

[0107] The color developer is iodine or phosphomolybdic acid in ethanol.

[0108] All extraction solvents were dried over anhydrous Na2SO4 unless otherwise specified.

[0109] 1 H-NMR was recorded using a Varian-300 nuclear magnetic resonance spectrometer with TMS as the internal standard.

[0110] LC-MS was recorded using an Agilent 1100 high performance liquid chromatography-ion trap mass spectrometer (LC-MSDTrap), a diode array detector (DAD), detection wavelengths of 214 nm and 254 nm, and an ion trap mass spectrometer (ESI source).

[0111] The HPLC column was Agela Durashell C18 (4.6×50 mm, 3.5 μm); the mobile phase was 0.1% NH 4 HCO 3 aqueous solution: acetonitrile (5:95 to 95:5 in 5 minutes, v / v); the flow rate was 1.8 mL / min.

[0112] The following abbreviations may be used in this disclosure: NBS (N-bromosuccinimide); T3P (1-propylphosphonic acid cyclic anhydride); DMF (N,N-dimethylformamide); LDA (lithium diisopropylamide); DCE (1,2-dichloroethane); DCM (dichloromethane); TEA (triethylamine); EA (ethyl acetate); THF (tetrahydrofuran); TFA (trifluoroacetic acid); TKB (potassium tert-butoxide); DAST (diethylaminosulfur trifluoride); DMEDA (N,N-dimethylethylenediamine); EG (ethylene glycol); Xantphos (4,5-bis(diphenylmethane)). (1,1'-Bis(diphenylphosphino)ferrocene)palladium dichloride); Pd(OAc)2 (palladium acetate); DPPA (diphenylphosphinoyl azide); STAB (sodium triacetoxyborohydride); DMSO (dimethyl sulfoxide); DIBAL-H (diisobutylaluminum hydride); DCC (N,N'-dicyclohexylcarbodiimide).

[0113] [Synthesis of Intermediate P-1]

[0114] Synthesis of intermediate 1:

[0115] Dissolve 2-chloro-5-hydroxypyridine (12 g, 96 mmol) in concentrated sulfuric acid (150 mL). Add potassium nitrate (12 g, 116 mmol) portionwise at 0°C. After 2 hours of addition, slowly warm to room temperature and stir for 3 hours. After the reaction is complete, slowly add the reaction solution dropwise to ice water to precipitate a yellow solid, which is filtered to obtain yellow intermediate 1 (15.4 g, 91%). 1 H-NMR (300MHz, DMSO-d6): δ11.94(s,1H),7.76(d,J=8.4Hz,1H),7.71(d,J=8.4Hz,1H).

[0116] Synthesis of intermediate 2:

[0117] Intermediate 1 (14 g, 83 mmol) was dissolved in a mixed solvent of tetrahydrofuran (300 mL) / water (100 mL). Zinc powder (44 g, 669 mmol) and ammonium chloride (89 g, 1673 mmol) were added portionwise at 50°C. After stirring for 1 hour, the mixture was cooled to room temperature and diluted with ethyl acetate (200 mL). Water (150 mL) was added and the mixture was stirred, separated, and filtered. The organic phase was collected, dried over anhydrous sodium sulfate, and chromatographed on a silica gel column (petroleum ether / ethyl acetate = 2 / 1, v / v) to give a yellow intermediate 2 (11 g, 94%). 1 H-NMR (300MHz, DMSO-d6): δ9.72(s,1H),6.82(d,J=7.8Hz,1H),6.37(d,J=7.8Hz,1H),5.88(s,2H).LC-MS: m / z 145.1[M+H] + .

[0118] Synthesis of intermediate 3:

[0119] Intermediate 2 (23 g, 162 mmol) and potassium ethyl xanthate (39 g, 243 mmol) were dissolved in pyridine (100 mL), refluxed at 120 ° C for 4 hours, cooled to room temperature, and pyridine was removed by rotary evaporation. Ice water (100 mL) was added to the residue, and the pH was adjusted to 1 with 3N dilute hydrochloric acid. An off-white solid precipitated. The mixture was stirred for 1 hour and filtered to obtain an off-white intermediate 3 (29 g, 96%). 1 H-NMR (300MHz, DMSO-d6): δ7.91 (d, J=8.4Hz, 1H), 7.35 (d, J=8.4Hz, 1H). LC-MS: m / z187.1[M+H] + .

[0120] Synthesis of intermediate 4:

[0121] Intermediate 3 (29 g, 155 mmol) was dissolved in ethyl acetate (300 mL), potassium carbonate (43 g, 313 mmol) was added, and iodomethane (20 mL, 311 mmol) was added at room temperature. After stirring for 2 hours, water (200 mL) was added. The liquids were separated, and the organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to obtain a gray solid intermediate 4 (28 g, 91%). 1 H-NMR (300MHz, DMSO-d6): δ8.14(d,J=8.4Hz,1H),7.42(d,J=8.4Hz,1H),2.79(s,3H).

[0122] Synthesis of intermediate 5:

[0123] Intermediate 4 (2.0 g, 10 mmol) and tert-butyl piperazine-1-carboxylate (5.0 g, 50 mmol) were dissolved in tetrahydrofuran (70 mL) and stirred at 80°C overnight. The reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1, v / v) to give a white solid product 5 (3.0 g, 72%). 1 H-NMR (300MHz, CDCl3): δ7.37(d,J=8.1Hz,1H),6.93(d,J=8.1Hz,1H),3.77-3.67(m,4H),3.64-3.51(m,4H),1.48(s,9H).

[0124] Synthesis of intermediate 6:

[0125] Intermediate 5 (3.0 g, 8.8 mmol) was dissolved in dry acetonitrile (100 mL), and NBS (2.2 g, 12.3 mmol) was slowly added in portions under an ice-water bath. The mixture was stirred at room temperature for 5 hours, diluted with ethyl acetate (200 mL), washed with 1N sodium hydroxide (50 mL × 4) aqueous solution, dried over anhydrous sodium sulfate, and concentrated to obtain a brown solid 6 (3.0 g, 83%). 1 H-NMR (300MHz, CDCl3): δ7.68(s,1H),3.77-3.71(m,4H),3.62-3.55(m,4H),1.49(s,9H).

[0126] Synthesis of intermediate 7:

[0127] Intermediate 6 (1 g, 2.4 mmol), ethyl acrylate (4.2 g, 48 mmol), palladium acetate (54 mg, 0.24 mmol), tri(o-methylphenyl)phosphine (220 mg, 0.72 mmol), and triethylamine (726 mg, 7.2 mmol) were suspended in dry N,N-dimethylformamide (5 mL) and reacted in a microwave oven at 140°C for 2 hours under nitrogen protection. The reaction solution was diluted with water (10 mL), extracted with ethyl acetate (20 mL × 3), washed with saturated brine, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1, v / v) to obtain the yellow solid product 7 (460 mg, 46%). 1 H-NMR (300MHz, CDCl3): δ8.07(d,J=15.9Hz,1H),7.64(s,1H),6.30(d,J=15.9Hz,1H),4.33-4. 22(m,2H),3.82-3.71(m,4H),3.64-3.52(m,4H),1.49(s,9H),1.35(t,J=5.7Hz,3H).LC-MS:m / z 437.1[M+H]+ .

[0128] Synthesis of intermediate 8:

[0129] Intermediate 7 (450 mg, 1.0 mmol) was dissolved in a dichloromethane / methanol mixed solvent (30 mL / 3 mL), and ozone was introduced at -78 ° C for 5 minutes, then dimethyl sulfide (0.5 mL) was added to quench the reaction, and the mixture was brought to room temperature, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1, v / v) to give a light yellow solid product 8 (250 mg, 68%). 1 H-NMR (300MHz, CDCl3): δ10.37(s,1H),7.91(s,1H),3.88-3.73(m,4H),3.66-3.52(m,4H),1.48(s,9H).LC-MS:m / z 367.1[M+H] + .

[0130] Synthesis of intermediate 9:

[0131] Intermediate 8 (50 mg, 0.13 mmol) was dissolved in tert-butanol / water (5 mL / 5 mL), and 2-methyl-2-butene (91 mg, 1.3 mmol) was added at room temperature. Then, a solution of sodium dihydrogen phosphate dihydrate (202 mg, 1.3 mmol) and sodium chlorite (59 mg, 0.65 mmol) in water (5 mL) was slowly added. The mixture was stirred at room temperature for 1 hour, diluted with water (30 mL), and the pH was adjusted to 3-4 with 1N hydrochloric acid. The mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, washed with saturated brine (20 mL × 3), and concentrated to give 9 (40 mg, 77%) as a white solid. 1 H-NMR (300MHz, DMSO-d6): δ8.14(s,1H),3.78-3.63(m,4H),3.60-3.46(m,4H),1.44(s,9H).LC-MS:m / z 383.1[M+H] + .

[0132] Synthesis of intermediate 10:

[0133] Intermediate 9 (40 mg, 0.1 mmol) was dissolved in dry N,N-dimethylformamide (2 mL). Triethylamine (69 mg, 0.5 mmol) and 2-amino-6-(1-methyl-1H-pyrazol-4-yl)pyridine (34 mg, 0.2 mmol) were added. A 50% solution of T3P (159 mg, 0.5 mmol) in N,N-dimethylformamide was added with stirring. The mixture was microwaved at 100°C for 2 hours. The reaction mixture was poured into ice water (10 mL), and the product precipitated. After stirring for 30 minutes, the product was filtered, washed with water, and dried to obtain a pale yellow solid 10 (10 mg, 18%). 1 H-NMR (300MHz, DMSO-d6): δ10.92(s,1H),8.18(s,1H),8.08(s,1H),7.98-7.91(m,2H),7.85-7 .75(m,1H),7.46-7.35(m,1H),3.87(s,3H),3.75-3.63(m,4H),3.57-3.48(m,4H),1.43(s,9H).

[0134] Synthesis of intermediate 11:

[0135] Intermediate 10 (80 mg, 0.15 mmol), p-methoxypiperidine (172 mg, 1.5 mmol) and potassium carbonate (103 mg, 0.75 mmol) were dissolved in dry 1,4-dioxane (5 mL) and stirred at 110 ° C for 24 hours. The reaction solution was filtered, the filtrate was concentrated, and purified by preparative plate chromatography (dichloromethane / methanol = 20 / 1, v / v) to give a white solid 11 (45 mg, 49%). 1 H-NMR (300MHz, DMSO-d6): δ13.46-13.31(m,1H),8.35-8.20(m,2H),8.10-7.99(m,2H),7.78(s,1H),7.39(s,1H), 3.87(s,3H),3.76-3.63(m,9H),3.25-3.13(m,5H),3.10-2.93(m,2H),2.25-1.98(m,4H),1.43(s,9H).LC-MS:m / z 618.3[M+H] + .

[0136] Synthesis of final product P-1:

[0137] Intermediate 11 (30 mg, 0.05 mmol) was dissolved in 2.8 N hydrochloric acid ethyl acetate solution (5 mL) and stirred at room temperature overnight. The reaction solution was concentrated, and dichloromethane (20 mL × 4) and water (20 mL) were added and back-extracted 4 times. The aqueous phase was taken and the pH was adjusted to 7-8 with saturated sodium bicarbonate aqueous solution under ice-water bath. Dichloromethane (30 mL × 3) was added for extraction. The organic phases were combined and concentrated to obtain P-1 (21 mg, 81%). 1 H NMR (300MHz, DMSO-d6): δ13.42(s,1H),8.26(s,1H),8.22(s,1H),8.08-8.0 0(m,2H),7.78(t,J=7.8Hz,1H),7.38(d,J=7.5Hz,1H),3.87(s,3H),3.66-3 .56(m,4H),3.48-3.43(m,1H),3.38(s,3H),3.23-3.17(m,3H),3.04-2.94( m,2H),2.88-2.75(m,4H),2.22-2.12(m,2H),2.10-1.94(m,2H).LC-MS:m / z 517.8[M+H] + .

[0138] Preparation Example 1

[0139] The synthesis of compound A1 was carried out by the following method:

[0140] Synthesis of intermediate A1-1:

[0141] Dissolve diglycolamine (2.6 g, 25 mmol) and triethylamine (2.6 g, 25 mmol) in dry dichloromethane (25 mL). Add a solution of benzyl chloroformate (4.3 g, 25 mmol) in dichloromethane (30 mL) slowly dropwise to the reaction system at 0°C. Stir the reaction mixture at 0°C for 2 hours and then at room temperature for 18 hours. Quench the reaction mixture with saturated NaHCO₃ solution (30 mL) and extract with dichloromethane three times (40 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain crude product A1-1 (4.7 g, 79%), which is used directly in the next step.

[0142] Synthesis of intermediate A1-2:

[0143] Intermediate A1-1 (500 mg, 2.1 mmol) and potassium tert-butoxide (255 mg, 2.1 mmol) were dissolved in dry tetrahydrofuran (10 mL). The reaction was stirred at 0°C for 0.5 hours, and then tert-butyl bromoacetate (490 mg, 2.5 mmol) was slowly added dropwise to the reaction system. The reaction was stirred at 0°C for 3 hours, then warmed to room temperature and stirred for 15 hours. The reaction was quenched by the addition of water (10 mL) and extracted three times with ethyl acetate (15 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified on a silica gel column (petroleum ether / ethyl acetate = 20 / 1, v / v) to afford A1-2 (210 mg, 28%) as a pale yellow oil.

[0144] Synthesis of intermediate A1-3:

[0145] Intermediate A1-2 (210 mg, 0.59 mmol) was dissolved in dry methanol (10 mL). Pd / C (200 mg) was added and stirred at room temperature under a hydrogen atmosphere for 12 hours. The organic phase was filtered and concentrated under reduced pressure to afford crude colorless oil A1-3 (180 mg, 100%), which was used directly in the next reaction.

[0146] Synthesis of intermediate A1-4:

[0147] Intermediate A1-3 (665 mg, 2.4 mmol) and 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (633 mg, 2.9 mmol, commercially available) were dissolved in dry DMF (10 mL), and DIPEA (622 mg, 4.8 mmol) was added. The mixture was stirred at 90°C for 12 hours. The mixture was extracted three times with ethyl acetate (15 mL × 3) and washed three times with saturated aqueous NaCl solution (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified on a silica gel column (petroleum ether / ethyl acetate = 1 / 1, v / v) to obtain A1-4 (210 mg, 15%) as a yellow solid.

[0148] Synthesis of intermediate A1-5:

[0149] Intermediate A1-4 (110 mg, 0.23 mmol) was dissolved in dry dichloromethane (5 mL), trifluoroacetic acid (4 mL) was added, and the mixture was stirred at room temperature for 12 hours. The organic phase was concentrated under reduced pressure, and dry dichloromethane (5 mL) was added to the residue, which was concentrated under reduced pressure again to obtain the crude product, which was used directly in the next reaction.

[0150] Synthesis of final product A1:

[0151] Intermediate A1-5 (97 mg, 0.23 mmol), intermediate P-1 (63 mg, 0.23 mmol), HATU (43 mg, 0.35 mmol), and DIPEA (300 mg, 2.32 mmol) were dissolved in dry dichloromethane (3 mL) and stirred at room temperature for 12 hours. Water (3 mL) was added to the reaction solution, which was then extracted three times with ethyl acetate (10 mL x 3) and washed three times with saturated NaCl solution (15 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A1 (120 mg, 57%).

[0152] Preparation Example 2

[0153] The synthesis of compound A2 was carried out by the following method:

[0154] Synthesis of intermediate A2-1:

[0155] P-1 (1.0 g, 1.9 mmol) and N-tert-butyloxycarbonyl-4-piperidone (950 mg, 4.8 mmol) were suspended in dry 1,2-dichloroethane (30 mL) and stirred at room temperature for 10 minutes. Sodium triacetoxyborohydride (1.0 g, 4.8 mmol) was then added and allowed to react at room temperature for 12 hours. After completion, the reaction was quenched with saturated sodium bicarbonate solution (30 mL). The mixture was extracted with dichloromethane (50 mL × 2), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and spin-dried. The residue was added with diethyl ether (40 mL) and slurried for 8 hours. The mixture was filtered, rinsed with diethyl ether, and dried to obtain A2-1 (1.1 g, 81%) as a white solid. 1 H NMR (300MHz, DMSO-d6): δ13.42(s,1H),8.30-8.23(m,2H),8.08-8.02(m,2H),7.82-7.75(m,1 H),7.42-7.35(m,1H),3.99-3.91(m,2H),3.88(s,3H),3.67(s,3H),3.49-3.41(m,2H),3.40-3 .34(m,4H),3.24-3.17(m,2H),3.05-2.96(m,2H),2.74-2.66(m,2H),2.65-2.58(m,4H),2.21- 2.13(m,2H),2.10-2.00(m,2H),1.76-1.67(m,2H),1.39(s,9H),1.32-1.22(m,2H).LC-MS:m / z 700.8[M+H] + .

[0156] Synthesis of intermediate A2-2:

[0157] A2-1 (1.1 g, 1.6 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 8 hours. After the reaction was completed, the solvent was dried by rotary evaporation, diluted with water (10 mL), and saturated sodium bicarbonate was added dropwise to adjust the pH to 7-8. The mixture was extracted with dichloromethane (50 mL × 2), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and rotary evaporation to obtain a light yellow solid A2-2 (860 mg, 91%). 1 H NMR (300MHz, DMSO-d6): δ13.43(s,1H),8.30-8.24(m,2H),8.09-8.02(m,2H), 7.84-7.76(m,1H),7.43-7.35(m,1H),3.88(s,3H),3.68(s,3H),3.50-3.40(m, 4H),3.28-3.17(m,5H),3.03-2.93(m,4H),2.66-2.59(m,4H),2.44-2.36(m,2H ),2.21-2.14(m,2H),2.11-2.02(m,2H),1.72-1.63(m,2H),1.34-1.22(m,2H).

[0158] Synthesis of final product A2:

[0159] A2-2 (40 mg, 0.068 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (23 mg, 0.081 mmol) were dissolved in DMSO (2 mL). DIPEA (45 mg, 0.35 mmol) was added and the mixture was reacted at 95°C for 6 hours. Upon addition of water (5 mL), a large amount of solid precipitated. The solid was filtered and the filter cake was dried. The filter cake was purified by silica gel column chromatography (dichloromethane / methanol = 30 / 1, v / v) to yield product A2 (21 mg, 36%).

[0160] Preparation Example 3

[0161] The synthesis of compound A3 was carried out by the following method:

[0162] Synthesis of intermediate A3-1:

[0163] P-1 (60 mg, 0.12 mmol) and triethylamine (23 mg, 0.057 mmol) were dissolved in dichloromethane (2.5 mL). Chloroacetyl chloride (17 mg, 0.038 mmol) was added dropwise under an ice-water bath and allowed to react for 2 hours. The reaction was quenched by adding water (10 mL) and extracted with dichloromethane (10 mL x 2). The organic phase was dried and concentrated to afford crude A3-1 (60 mg) as a white solid. 1 H NMR (300MHz, DMSO-d6): δ13.41(s,1H),8.32-8.23(m,2H),8.08-8.00(m,2H),7.80(t,J=7.8Hz,1H),7.40(d,J=7.5Hz,1H),4.48(s, 2H),3.88(s,3H),3.79-3.69(m,4H),3.68-3.60(m,4H),3.44-3.30(m,4H),3.26-3.15(m,2H),3.08-2.95(m,2H),2.24-1.98(m,4H).

[0164] Synthesis of intermediate A3-2:

[0165] A3-1 (40 mg, 0.070 mmol) and piperazine (29 mg, 0.35 mmol) were dissolved in DMA (5 mL). KI (5 mg, 0.030 mmol) was added at room temperature and allowed to react for 3 hours. Water (10 mL) was added to precipitate a white solid, which was filtered and dried to afford crude product A3-2 (40 mg).

[0166] Synthesis of final product A3:

[0167] A3-2 (40 mg, 0.062 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (23 mg, 0.074 mmol, commercially available) were dissolved in DMSO (2 mL). DIPEA (40 mg, 0.032 mmol) was added and the mixture was reacted at 95°C for 6 hours. Upon addition of water (5 mL), a large amount of solid precipitated. The solid was filtered and the filter cake was dried. The filter cake was purified by silica gel column chromatography (dichloromethane / methanol = 30 / 1, v / v) to yield product A3 (17 mg, 29%).

[0168] Preparation Example 4

[0169] The synthesis of compound A4 was carried out by the following method:

[0170] Synthesis of intermediate A4-1:

[0171] 1-tert-Butyloxycarbonyl-4-piperidinecarboxaldehyde (350 mg, 1.6 mmol) was dissolved in dry ethyl acetate (5 mL). 2.8N hydrochloric acid in ethyl acetate (10 mL) was added dropwise to the reaction mixture at 0°C. The reaction mixture was then stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, and dry dichloromethane (5 mL) was added to the residue. The pH of the solution was adjusted to 7 with solid sodium bicarbonate powder. The solution was filtered and concentrated under reduced pressure to obtain crude product A4-1 (185 mg, 99%), which was used directly in the next step.

[0172] Synthesis of intermediate A4-2:

[0173] 2-(2,6-Dioxypyridin-3-yl)-5-fluoroisoindoline-1,3-dione (293 mg, 1.1 mmol, commercially available) and intermediate A4-1 (180 mg, 1.6 mmol) were dissolved in dry dimethyl sulfoxide (5 mL). DIPEA (685 mg, 5.3 mmol) was added and stirred at 95°C for 12 hours. After dilution with water, the mixture was extracted three times with ethyl acetate (15 mL × 3) and washed three times with saturated NaCl solution (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 25 / 1, v / v) to afford A4-2 (55 mg, 14%) as a yellow solid. 1 H NMR (300MHz, DMSO-d6): δ11.08(s,1H),9.61(s,1H),7.65(d,J=6.0Hz,1H),7.48-7.12(m,2H),5.19-4.98(m,1H),4.16- 3.82(m,2H),3.29-3.07(m,2H),2.97-2.82(m,1H),2.77-2.52(m,4H),2.08-1.86(m,2H),1.72-1.42(m,2H).LC-MS:m / z 402.1[M+H+CH3OH] + .

[0174] Synthesis of final product A4:

[0175] Intermediate A4-2 (55 mg, 0.15 mmol) and intermediate P-1 (93 mg, 0.18 mmol) were dissolved in dry 1,2-dichloroethane (4 mL). One drop of acetic acid and activated molecular sieves (100 mg) were added, followed by stirring at room temperature for 1 hour. Sodium triacetoxyborohydride (63 mg, 0.30 mmol) was added to the reaction mixture, which was then stirred at room temperature for 12 hours. The reaction mixture was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A4 (12 mg, 9%).

[0176] Preparation Example 5

[0177] The synthesis of compound A5 was carried out by the following method:

[0178] Synthesis of intermediate A5-1:

[0179] P-1 (700 mg, 1.4 mmol), 1-(tert-butyloxycarbonyl)piperidine-4-carboxylic acid (372 mg, 1.7 mmol), triethylamine (411 mg, 4.1 mmol), and HATU (792 mg, 2.1 mmol) were sequentially dissolved in DMF (15 mL) and reacted at room temperature for 3 hours. Saturated brine (20 mL) was added, and the mixture was extracted with dichloromethane (30 mL x 2). The organic phase was dried, concentrated, and purified by silica gel column chromatography (dichloromethane / methanol = 40 / 1, v / v) to obtain A5-1 (800 mg, 78%) as a white solid. LC-MS: m / z 315.2 [(M-Boc+2H) / 2] 2+ .

[0180] Synthesis of intermediate A5-2:

[0181] A5-1 (200 mg, 0.27 mmol) was dissolved in dichloromethane (5 mL). Trifluoroacetic acid (1.2 g, 11 mmol) was added at room temperature and allowed to react for 3 hours. The reaction solution was concentrated, and a dichloromethane / methanol mixture (5 mL) was added. Saturated sodium bicarbonate aqueous solution was added dropwise under an ice-water bath to adjust the pH to 8. The solution was extracted with dichloromethane (20 mL x 3) and concentrated to afford A5-2 (60 mg, 36%) as a white solid. 1 H NMR (300MHz, DMSO-d6): δ13.42(s,1H),8.30(s,1H),8.28(s,1H),8.08-8.01(m,2H),7.80(t,J=7.5Hz,1H),7.40(d,J=7.5Hz,1H),3.88 (s,3H),3.75-3.62(m,8H),3.22(s,3H),3.06-2.98(m,4H),2.95-2.87(m,4H),2.75-2.66(m,2H),2.25-1.89(m,5H),1.51-1.48(m,4H).

[0182] Synthesis of final product A5:

[0183] A5-2 (60 mg, 0.096 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (31 mg, 0.14 mmol, commercially available) were dissolved in DMSO (2 mL). DIPEA (62 mg, 0.078 mmol) was added and the mixture was reacted at 95°C for 6 hours. Upon addition of water (5 mL), a large amount of solid precipitated. The solid was filtered and the filter cake was dried. The filter cake was purified by silica gel column chromatography (dichloromethane / methanol = 30 / 1, v / v) to yield product A5 (42 mg, 29%).

[0184] Preparation Example 6

[0185] The synthesis of compound A6 was carried out by the following method:

[0186] Synthesis of intermediate A6-1:

[0187] Methyl 1-(tert-butoxycarbonyl)azetidine-3-carboxylate (800 mg, 3.7 mmol) was dissolved in dry DCM (5 mL). Under nitrogen, 1N DIBAL-H tetrahydrofuran solution (8.2 mL, 8.2 mmol) was added dropwise to the reaction mixture at -78°C. The mixture was then stirred at -78°C for 0.5 hours. The reaction mixture was slowly warmed to 0°C and anhydrous methanol (2 mL) was added to stop the reaction. 10% aqueous citric acid solution (30 mL) was added to the reaction mixture, resulting in the appearance of a white solid. The reaction mixture was stirred at 0°C until the white solid disappeared. The mixture was extracted three times with dichloromethane (15 mL x 3), the organic phases were combined, and the organic phases were dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (petroleum ether / ethyl acetate = 20 / 1, v / v) to obtain A6-1 (220 mg, 32%) as a colorless oil. 1 H NMR (300MHz, CDCl3): δ9.84(s,1H),4.14-3.94(m,2H),3.81-3.64(m,2H),3.52-3.44(m,1H),1.43(s,9H).

[0188] Synthesis of intermediate A6-2:

[0189] Intermediate P-1 (186 mg, 0.36 mmol) and intermediate A6-1 (200 mg, 1.1 mmol) were dissolved in dry 1,2-dichloroethane (4 mL). One drop of acetic acid and activated molecular sieves (200 mg) were added, followed by stirring at room temperature for 1 hour. Sodium triacetoxyborohydride (229 mg, 1.1 mmol) was added to the reaction mixture, which was then stirred at room temperature for 12 hours. The reaction mixture was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 30 / 1, v / v) to afford A6-2 (140 mg, 53%) as a white solid. 1 H NMR (300MHz, CDCl3): δ13.46(s,1H),8.37(s,1H),8.17(s,1H),8.14(d,J=7.5Hz,1H),7.93(s,1 H),7.68(t,J=7.8Hz,1H),7.21(d,J=7.5Hz,1H),4.10-3.99(m,2H),3.96(s,3H),3.89-3.70(m,4 H),3.67-3.54(m,2H),3.47(s,3H),3.45-3.39(m,1H),3.39-3.26(m,2H),3.25-3.07(m,2H),2.8 1-2.70(m,1H),2.70-2.61(m,2H),2.60-2.47(m,4H),2.39-2.13(m,4H),1.44(s,9H).LC-MS:m / z 687.4[M+H] + .

[0190] Synthesis of intermediate A6-3:

[0191] Intermediate A6-2 (140 mg, 0.21 mmol) was dissolved in dry ethyl acetate (2 mL), and 2.8 N hydrochloric acid in ethyl acetate (2 mL) was added. The mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, and dichloromethane (10 mL) and methanol (2 mL) were added to the residue. The pH was adjusted to 7 with solid NaHCO3 powder, filtered, and dried to afford A6-3 (120 mg, 100%) as a white solid. 1H NMR (300MHz, CDCl3): δ13.45(s,1H),8.37(s,1H),8.26-8.06(m,2H),7.93(s,1H),7. 68(t,J=7.5Hz,1H),7.21(d,J=7.2Hz,1H),3.96(s,3H),3.90-3.70(m,4H),3.69-3.54 (m,2H),3.53-3.45(m,4H),3.40-3.26(m,2H),3.25-3.13(m,2H),3.09-2.89(m,6H),2 .75-2.66(m,1H),2.64-2.48(m,3H),2.29-2.18(m,2H),2.09-1.98(m,2H).LC-MS:m / z 587.3[M+H] + .

[0192] Synthesis of final product A6:

[0193] Intermediate A6-3 (60 mg, 0.10 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (37 mg, 0.13 mmol, commercially available) were dissolved in dry dimethyl sulfoxide (2 mL). DIPEA (66 mg, 0.51 mmol) was added, and the mixture was stirred at 95°C for 12 hours. Water (3 mL) was added to the mixture, and a yellow solid precipitated. The filter cake was filtered and purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A6 (50 mg, 58%).

[0194] Preparation Example 7

[0195] The synthesis of compound A7 was carried out by the following method:

[0196] Synthesis of intermediate A7-1:

[0197] 5-Bromo-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (2.0 g, 5.9 mmol, commercially available), potassium ethylene trifluoroborate (2.4 g, 17.8 mmol), potassium carbonate (1.6 g, 11.8 mmol), and Pd(dppf)Cl2 (428 mg, 0.59 mmol) were suspended in 1,4-dioxane (60 mL). After nitrogen replacement, the mixture was reacted at 100°C overnight. The reaction solution was concentrated and purified by silica gel column chromatography to obtain A7-1 (1.0 g, 62%) as an orange solid. 1H NMR (300MHz, DMSO-d6): δ11.13(s,1H),8.07(s,1H),7.99-7.93(m,1H),7.92-7.86(m,1H),6.95(dd,J=17.4,10.8Hz,1H),6.20(d,J= 17.4Hz,1H),5.54(d,J=10.8Hz,1H),5.21-5.10(m,1H),2.98-2.81(m,1H),2.65-2.60(m,1H),2.59-2.54(m,1H),2.13-2.00(m,1H).

[0198] Synthesis of intermediate A7-2:

[0199] A7-1 (400 mg, 1.4 mmol) was dissolved in 1,4-dioxane / water (20 mL / 10 mL). Potassium osmate (52 mg, 0.14 mmol), sodium periodate (1.2 g, 5.6 mmol), and 2,6-lutidine (300 mg, 2.8 mol) were added in one portion under an ice-water bath and allowed to react for 5 hours. Sodium thiosulfate pentahydrate solution (10 mL) was added, and the mixture was extracted with dichloromethane (10 mL × 3). The product A7-2 (80 mg, 20%) was obtained as a yellow solid by silica gel column chromatography. LC-MS: m / z 285.1 [MH] - .

[0200] Synthesis of final product A7:

[0201] A7-2 (33 mg, 0.12 mmol) and A5-2 (87 mg, 0.15 mmol) were dissolved in dry 1,2-dichloroethane (4 mL), and tetraisopropyl titanate (49 mg, 0.17 mmol) was added. After 2 hours, sodium triacetoxyborohydride (96 mg, 0.46 mmol) was added and allowed to react overnight. The reaction was quenched by the addition of saturated ammonium chloride solution (6 mL), extracted with dichloromethane (20 mL × 3), and purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1, v / v) to obtain A7 (26 mg, 24%).

[0202] Preparation Example 8

[0203] The synthesis of compound A8 was carried out by the following method:

[0204] Synthesis of intermediate A8-1:

[0205] P-1 (100 mg, 0.19 mmol) was dissolved in acetone (8 mL), and 1-bromo-2-chloroethane (42 mg, 0.29 mmol) and potassium carbonate (52 mg, 0.38 mmol) were added. The mixture was stirred at 60°C for 96 hours. The reaction mixture was then dried and purified by silica gel column chromatography to obtain intermediate A8-1 (80 mg, 73%) as a white solid. 1 H NMR (300MHz, DMSO-d6): δ13.43(s,1H),8.35-8.20(m,2H),8.13-7.96(m,2H),7.88-7.76(m,1H),7.45-7.35(m,1H) ,3.87(s,3H),3.83-3.59(m,5H),3.30-3.12(m,6H),3.01(s,3H),2.84-2.55(m,6H),2.23-1.98(m,4H).LC-MS:m / z 580.2[M+H] + .

[0206] Synthesis of intermediate A8-2:

[0207] Intermediate A8-1 (60 mg, 0.10 mmol) was dissolved in dry N,N-dimethylformamide (8 mL). Piperazine (45 mg, 0.52 mmol) and potassium carbonate (71 mg, 0.52 mmol) were added, and the mixture was stirred at 80°C for approximately 5 hours. The reaction solution was cooled to room temperature, water (15 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The mixture was concentrated to obtain a pale yellow solid crude product A8-2 (65 mg, 100%), which was used directly in the next reaction. LC-MS: m / z 630.4 [M+H] + .

[0208] Synthesis of final product A8:

[0209] Intermediate A8-2 (50 mg, 0.080 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (29 mg, 0.11 mmol, commercially available) were dissolved in dimethyl sulfoxide (2 mL). N,N-diisopropylethylamine (52 mg, 0.40 mmol) was added, and the mixture was stirred at 95°C overnight. The reaction solution was cooled to room temperature, and water (10 mL) was added to precipitate a solid. The solid was filtered, and the filter cake was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1, v / v) to obtain A8 (23 mg, 33%).

[0210] Preparation Example 9

[0211] The synthesis of compound A9 was carried out by the following method:

[0212] Synthesis of intermediate A9-1:

[0213] Methyl 4-bromo-2-bromomethylbenzoate (450 mg, 1.5 mmol), 3-amino-2,6-piperidinedione (206 mg, 1.6 mmol), and potassium carbonate (605 mg, 4.4 mmol) were dissolved in dry N,N-dimethylformamide (8 mL) and stirred at 70°C for 16 hours. The reaction solution was concentrated under reduced pressure, and water (10 mL) was added to the residue, which was slurried for 30 minutes, filtered, and washed with anhydrous ether to obtain Intermediate A9-1 (200 mg, 42%). 1 H NMR (300MHz, DMSO-d6): δ11.00(s,1H),7.89(s,1H),7.77-7.57(m,2H),5.16-5.05(m,1H),4.52-4 .29(m,2H),2.96-2.83(m,1H),2.68-2.59(m,1H),2.43-2.26(m,1H),2.08-1.95(m,1H).LC-MS:m / z 323.0[M+H] + .

[0214] Synthesis of intermediate A9-2:

[0215] Intermediate A9-1 (200 mg, 0.62 mmol), DCC (26 mg, 0.12 mmol), triethylamine (13 mg, 0.12 mmol), formic acid (100 mg, 2.2 mmol), palladium acetate (7.0 mg, 0.030 mmol), and Xantphos (18 mg, 0.030 mmol) were dissolved in dry N,N-dimethylformamide (8 mL) and stirred at 110°C under nitrogen for 3 hours. The reaction mixture was filtered and concentrated under reduced pressure. Dry dichloromethane (5 mL) was added to the residue and the mixture was slurried. The solution was filtered to obtain Intermediate A9-2 (90 mg, 50%). 1 H NMR (300MHz, DMSO-d6): δ11.03(s,1H),8.17(s,1H),8.07(d,J=7.8Hz,1H),7.83(d,J=7.8Hz,1H),5.21-5.08(m ,1H),4.59-4.34(m,2H),2.96-2.83(m,1H),2.67-2.55(m,1H),2.45-2.31(m,1H),2.09-1.96(m,1H).LC-MS:m / z 289.1[M+H] + .

[0216] Synthesis of final product A9:

[0217] Intermediate A9-2 (40 mg, 0.14 mmol), intermediate A2-2 (83 mg, 0.14 mmol), HATU (53 mg, 0.14 mmol), and DIPEA (108 mg, 0.83 mmol) were dissolved in dry N,N-dimethylformamide (2 mL) and stirred at room temperature for 16 hours. Water (3 mL) was added to the reaction solution, which was then extracted three times with ethyl acetate (10 mL × 3) and washed three times with saturated aqueous NaCl (15 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield the final product A9 (5 mg, 4%).

[0218] Preparation Example 10

[0219] The synthesis of compound A11 was carried out by the following method:

[0220] Synthesis of intermediate A11-1:

[0221] Intermediate 9 (200 mg, 0.52 mmol) and 2-amino-6-methoxypyridine (98 mg, 0.79 mmol) were dissolved in DMA (0.4 mL). DIPEA (335 mg, 2.6 mmol) and a 50% T3P solution in ethyl acetate (1.6 g, 2.6 mmol) were added, and the mixture was microwaved at 110°C for 2 hours. After completion of the reaction, the EA was evaporated, the mixture was slurried with water (10 mL), filtered, and dried to obtain A11-1 (210 mg, 82%) as a brown solid. 1 H NMR (300MHz, DMSO-d6): δ10.87(s,1H),8.06(s,1H),7.80-7.69(m,2H),6.61- 6.55(m,1H),3.81(s,3H),3.72-3.66(m,4H),3.54-3.49(m,4H),1.43(s,9H).

[0222] Synthesis of intermediate A11-2:

[0223] A11-1 (210 mg, 0.43 mmol) and 4-methoxypiperidine (390 mg, 3.4 mmol) were dissolved in 1,4-dioxane (1.5 mL), potassium carbonate (297 mg, 2.2 mmol) was added and reacted at 105 ° C for 16 hours. After the reaction was completed, the mixture was spin-dried, diluted with water (5 mL), extracted with DCM (10 mL×2), dried over anhydrous sodium sulfate, spin-dried, slurried with ether (10 mL) for 8 hours, filtered, and dried to obtain a light yellow solid crude product A11-2 (170 mg).

[0224] Synthesis of intermediate A11-3:

[0225] The crude product of A11-2 (170 mg, 0.29 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added to react at room temperature for 6 hours. After the reaction was completed, the mixture was dried by rotary evaporation, diluted with water (15 mL), and extracted with DCM (10 mL × 2) to obtain an aqueous phase. The pH value was adjusted to 7-8 with saturated sodium bicarbonate solution, extracted with DCM (20 mL × 2), dried over anhydrous sodium sulfate, and dried by rotary evaporation to obtain a white solid A11-3 (130 mg, 65%). 1 H NMR (300MHz, DMSO-d6): δ13.23(s,1H),8.22(s,1H),7.84(d,J=7.8Hz,1H),7.72(t,J=7.8Hz,1H),6.55(d,J=7.8Hz,1H),3.84(s,3 H),3.64-3.58(m,5H),3.28(s,3H),3.22-3.16(m,2H),2.99-2.91(m,2H),2.83-2.78(m,4H),2.17-2.09(m,2H),1.98-1.88(m,2H).

[0226] Synthesis of intermediate A11-4:

[0227] A11-3 (130 mg, 0.28 mmol) was dissolved in DCE (5 mL), and A6-1 (130 mg, 0.71 mmol) was added. The mixture was reacted at room temperature for half an hour. STAB (150 mg, 0.71 mmol) was added and the mixture was reacted for 8 hours. After the reaction was completed, saturated sodium bicarbonate (10 mL) was added to quench the reaction. The mixture was extracted with DCM (15 mL × 2), dried over anhydrous sodium sulfate, and dried by spin drying. The mixture was purified by silica gel column chromatography (DCM / MeOH = 25 / 1, v / v) to give A11-4 (108 mg, 61%) as a white solid. 1H NMR (300MHz, DMSO-d6): δ13.20(s,1H),8.22(s,1H),7.84(d,J=7.8Hz,1H),7.72(t,J=7 .8Hz,1H),6.55(d,J=7.8Hz,1H),3.95-3.88(m,2H),3.84(s,3H),3.70-3.64(m,4H),3.5 2-3.47(m,2H),3.37-3.34(m,5H),3.29(s,3H),3.23-3.16(m,2H),3.00-2.90(m,2H),2 .80-2.71(m,1H),2.60-2.55(m,2H),2.16-2.08(m,2H),1.98-1.87(m,2H),1.37(s,9H).

[0228] Synthesis of intermediate A11-5:

[0229] Intermediate A11-4 (108 mg, 0.17 mmol) was dissolved in dry dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, and dichloromethane (10 mL) and methanol (2 mL) were added to the residue. The pH was adjusted to 7 with solid NaHCO3 powder, and the filtrate was filtered and dried to give crude intermediate A11-5 (90 mg, 99%). 1 H NMR (300MHz, DMSO-d6): δ13.21(s,1H),8.23(s,1H),7.84(d,J=7.2Hz,1H),7.7 2(t,J=7.2Hz,1H),6.54(d,J=7.5Hz,1H),4.06-3.94(m,2H),3.84(s,3H),3.73- 3.63(m,4H),3.47-3.36(m,10H),3.30-3.26(m,3H),3.02-2.90(m,2H),2.67-2 .60(m,1H),2.58-2.51(m,2H),2.22-2.05(m,2H),2.03-1.85(m,2H).LC-MS:m / z 537.0[M+H] + .

[0230] Synthesis of final product A11:

[0231] Intermediate A11-5 (90 mg, 0.17 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (60 mg, 0.22 mmol, commercially available) were dissolved in dry dimethyl sulfoxide (2 mL). DIPEA (108 mg, 0.84 mmol) was added, and the mixture was stirred at 95°C for 12 hours. Water (3 mL) was added to the mixture, and a yellow solid precipitated. The residue was filtered and purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A11 (100 mg, 75%).

[0232] Preparation Example 11

[0233] The synthesis of compound A12 was carried out by the following method:

[0234] Synthesis of intermediate A12-1:

[0235] A45-11 (120 mg, 0.28 mmol) and triethylamine (56 mg, 0.55 mmol) were dissolved in dry dichloromethane (3 mL). Chloroacetyl chloride (41 mg, 0.36 mmol) was added in an ice-water bath and the mixture was stirred at room temperature for 2 hours. Water (2 mL) was added to the reaction system to quench the reaction. The mixture was then extracted three times with 30 mL of dichloromethane (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A12-1 (120 mg, 85%) as a brown solid. 1 H NMR (300MHz, DMSO-d6): δ10.40(s,1H),8.36-8.22(m,2H),8.10-7.98(m,2H),7.80(t,J=7.8Hz,1H),7.41(d,J= 7.5Hz,1H),4.48(s,2H),4.16(s,3H),3.90(s,3H),3.82-3.69(m,4H),3.61-3.48(m,4H).LC-MS:m / z510.8[M+H] + .

[0236] Synthesis of intermediate A12-2:

[0237] A12-1 (80 mg, 0.16 mmol), piperazine (67 mg, 0.78 mmol), and potassium iodide (2 mg, 0.010 mmol) were dissolved in dry N,N-dimethylacetamide (2 mL) and stirred at room temperature for 3 hours. 2 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The mixture was washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A12-2 (30 mg, 34%) as a brown solid. LC-MS: m / z 560.9 [M+H] + .

[0238] Synthesis of final product A12:

[0239] Intermediate A12-2 (30 mg, 0.05 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (19 mg, 0.070 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (35 mg, 0.27 mmol) was added, and the mixture was stirred at 90°C for 12 hours. Water (3 mL) was added to the mixture, and a yellow solid precipitated. The residue was filtered and purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A12 (8 mg, 18%).

[0240] Preparation Example 12

[0241] The synthesis of compound A13 was carried out by the following method:

[0242] Synthesis of intermediate A13-1:

[0243] A45-11 (150 mg, 0.35 mmol), 2-bromoethanol (430 mg, 3.5 mmol), and potassium carbonate (190 mg, 1.4 mmol) were dissolved in DMF (4 mL) and reacted at 80°C for 6 hours. After completion of the reaction, the mixture was extracted with water (10 mL) and dichloromethane (20 mL × 2). The mixture was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and dried. The residue was slurried with diethyl ether (10 mL), filtered, and dried to obtain A13-1 (81 mg, 49%) as a white solid. LC-MS: m / z 478.9 [M+H] + .

[0244] Synthesis of intermediate A13-2:

[0245] Oxalyl chloride (35 mg, 0.28 mmol) was dissolved in DCM (0.5 mL) and allowed to stand at -78°C for 5 minutes. DMSO (43 mg, 0.55 mmol) was then added. After 30 minutes, A13-1 (30 mg, 0.062 mmol) was added, followed by triethylamine (86 mg, 0.85 mmol) 30 minutes later. The reaction was continued for 30 minutes, then brought to room temperature and allowed to react for 1.5 hours. After completion, the reaction was quenched with saturated sodium bicarbonate solution (3 mL) and extracted with dichloromethane (15 mL x 2). The product was washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, filtered, and dried. The product was slurried with diethyl ether (5 mL), filtered, and dried to obtain A13-2 (20 mg, 67%) as a pale yellow solid. LC-MS: m / z 508.9 [M+MeOH+H] + .

[0246] Synthesis of intermediate A13-3:

[0247] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (150 mg, 0.54 mmol) was dissolved in DMSO (5 mL), and 1-(tert-butyloxycarbonyl)piperazine (112 mg, 0.60 mmol) and DIPEA (350 mg, 2.7 mmol) were added and reacted at 90 ° C overnight. After the reaction was completed, water (10 mL) and dichloromethane (20 mL×2) were added for extraction, washed with saturated sodium chloride solution (5 mL×3), dried over anhydrous sodium sulfate, filtered, and dried. The product was slurried with ether (10 mL), filtered, and dried to obtain a yellow solid crude product (170 mg). The crude product was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for two hours. After the reaction was completed, the solvent was dried to obtain a yellow solid crude product A13-3 (190 mg). LC-MS: m / z 342.9[M+H] + .

[0248] Synthesis of final product A13:

[0249] A13-3 (25 mg, 0.054 mmol) was dissolved in DCE (0.5 mL), and triethylamine (18 mg, 0.18 mmol) was added. After 30 minutes, A13-2 (20 mg, 0.042 mmol) was added, and sodium triacetoxyborohydride (23 mg, 0.11 mmol) was added after another 30 minutes. The reaction was allowed to react overnight. After completion of the reaction, saturated sodium bicarbonate solution (3 mL) was added to quench the reaction and extracted with dichloromethane (10 mL × 2). The mixture was washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. A13 (6 mg, 18%) was obtained by preparative chromatography on a silica gel plate (dichloromethane / methanol = 15 / 1, v / v).

[0250] Preparation Example 13

[0251] The synthesis of compound A35 was carried out by the following method:

[0252] Synthesis of intermediate A35-1:

[0253] Methyl 2-cyano-4-fluorobenzoate (2.6 g, 14.5 mmol), 3-azetidinemethanol hydrochloride (2.0 g, 16.2 mmol), and potassium carbonate (8.0 g, 58.0 mmol) were dissolved in DMSO (35 mL) and reacted at 110°C overnight. After completion, the reaction was diluted with ethyl acetate (40 mL), followed by water (100 mL). The mixture was extracted with ethyl acetate (100 mL x 2), washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was purified by silica gel column chromatography (dichloromethane / ethyl acetate = 5 / 1, v / v) to obtain A35-1 (1.2 g, 34%) as a white solid. LC-MS: m / z 246.9 [M+H] + .

[0254] Synthesis of intermediate A35-2:

[0255] A35-1 (1 g, 4.1 mmol) and sodium hypophosphite monohydrate (4.3 g, 41 mmol) were dissolved in water (4 mL). Acetic acid (4 mL) and pyridine (8 mL) were added and the mixture was incubated at 70°C. Raney nickel (1 g) was added and the mixture was allowed to react for 8 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (40 mL x 2). The organic phase was washed twice with 1N hydrochloric acid (20 mL x 2) and once with saturated sodium chloride (20 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and dried to give crude A35-2 (800 mg) as a yellow solid. LC-MS: m / z 249.9 [M+H] + .

[0256] Synthesis of intermediate A35-3:

[0257] The crude product of A35-2 (800 mg) was dissolved in methanol (20 mL), and 3-amino-2,6-piperidinedione (240 mg, 1.9 mmol) and glacial acetic acid (1.0 g, 17.3 mmol) were added. The mixture was stirred at room temperature. After 30 minutes, sodium cyanoborocyanide (220 mg, 3.5 mmol) was added, and the mixture was allowed to react at 35°C for 16 hours. After completion of the reaction, the reaction solution was poured into ice water (30 mL) and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. A solid precipitated and was rinsed with diethyl ether (10 mL). The solid was slurried with ethyl acetate (10 mL), filtered, and dried to obtain a dark blue solid (240 mg, 46%). LC-MS: m / z 330.0 [M+H] + .

[0258] Synthesis of intermediate A35-4:

[0259] Oxalyl chloride (191 mg, 1.5 mmol) was dissolved in DCM (8 mL) and allowed to stand at -78°C. After 5 minutes, DMSO (235 mg, 3.0 mmol) was added. After 30 minutes, A15-3 (100 mg, 0.30 mmol) was added. After another 30 minutes, triethylamine (455 mg, 4.5 mmol) was added. The reaction was continued for 30 minutes, then brought to room temperature and continued for 1.5 hours. After completion, the reaction was quenched by adding saturated sodium bicarbonate solution (8 mL) and extracted with dichloromethane (25 mL × 2). The product was washed with saturated sodium chloride solution (15 mL), dried over anhydrous sodium sulfate, filtered, and dried to obtain crude A35-4 (100 mg) as a yellow oil. LC-MS: m / z 359.9 [M+MeOH+H] + .

[0260] Synthesis of final product A35:

[0261] The crude product of A35-4 (80 mg) was dissolved in DCE (2 mL), and P-1 (120 mg, 0.23 mmol) was added. After stirring at room temperature for 30 minutes, STAB (146 mg, 0.69 mmol) was added thereto and allowed to stand overnight. After the reaction was completed, saturated sodium bicarbonate solution (3 mL) was added to quench the reaction. The mixture was extracted twice with dichloromethane (20 mL × 2), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. A35 (10 mg, 5%) was obtained by preparative chromatography on a silica gel plate (dichloromethane / methanol = 15 / 1, v / v).

[0262] Preparation Example 14

[0263] The synthesis of compound A37 was carried out by the following method:

[0264] Synthesis of intermediate A37-1:

[0265] Potassium tert-butoxide (1.3 g, 11.8 mmol) was dissolved in dry tetrahydrofuran (40 mL). Triethyl phosphonoacetate (2.6 g, 11.8 mmol) was added at 0°C, and the mixture was slowly warmed to room temperature. Stirring was continued for 1 hour. Tert-butyl 6-oxo-2-azaspiro[3.3]heptane-2-carboxylate (1.0 g, 4.7 mmol) was then added at 0°C. The mixture was slowly warmed to room temperature and stirred for 16 hours. After completion, saturated sodium chloride solution (20 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1, v / v) to afford A37-1 (800 mg, 60%) as a yellow oil. LC-MS: m / z 304.1 [M+Na] + .

[0266] Synthesis of intermediate A37-2:

[0267] A37-1 (800 mg, 2.8 mmol) was dissolved in methanol (30 mL), 5% palladium on carbon (150 mg) was added, and a hydrogen balloon was added. The reaction was allowed to proceed overnight at room temperature. After the reaction was completed, the solution was filtered through celite and the filtrate was dried to obtain a yellow oily substance A37-2 (720 mg, 89%). 1 H NMR (300MHz, CDCl3): δ4.10 (q, J = 7.2Hz, 2H), 3.95-3.89 (m, 2H), 3.83-3.75 (m, 2H), 2.56 -2.46(m,1H),2.39-2.28(m,4H),1.91-1.81(m,2H),1.42(s,9H),1.23(t,J=7.2Hz,3H).

[0268] Synthesis of intermediate A37-3:

[0269] A37-2 (720 mg, 2.5 mmol) was dissolved in methanol (10 mL), and sodium hydroxide (500 mg, 12.5 mmol) was dissolved in water (10 mL). The mixture was added dropwise to the reaction mixture and allowed to react overnight at room temperature. After completion, the mixture was diluted with water (10 mL), concentrated, and the pH was adjusted to 4-5 with 1N hydrochloric acid. The mixture was extracted with DCM (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and dried to obtain A37-3 (630 mg, 97%) as a white solid. LC-MS: m / z 278.2 [M+Na] + .

[0270] Synthesis of intermediate A37-4:

[0271] A37-3 (100 mg, 0.40 mmol) was dissolved in tetrahydrofuran (2 mL), and P-1 (145 mg, 0.28 mmol), DIPEA (155 mg, 1.2 mmol), EDCI (77 mg, 0.40 mmol) and HOBT (54 mg, 0.40 mmol) were added in sequence. The mixture was reacted at room temperature overnight. After the reaction was completed, water (10 mL) was added to quench the mixture, and the mixture was extracted with dichloromethane (15 mL × 2) and dried over anhydrous sodium sulfate. The mixture was filtered, dried, and slurried with ether (10 mL), filtered, and dried to obtain a pale yellow solid crude product. The crude product was dissolved in dichloromethane (4 mL), and trifluoroacetic acid (2 mL) was added. The reaction was allowed to proceed at room temperature for 1 hour. After the reaction was complete, the solvent was dried, and a saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8. The product was extracted with dichloromethane (20 mL × 2), dried over anhydrous sodium sulfate, filtered, dried, slurried with ether (20 mL), filtered, and dried to obtain a pale yellow solid A37-4 (120 mg, 66%). LC-MS: m / z 654.9 [M+H] + .

[0272] Synthesis of final product A37:

[0273] A37-4 (50 mg, 0.080 mmol) was dissolved in dimethyl sulfoxide (1 mL), and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (28 mg, 0.10 mmol) and DIPEA (40 mg, 0.38 mmol) were added. The reaction was allowed to react at 90 ° C overnight. After the reaction was completed, dichloromethane (10 mL) was added for extraction, and the mixture was washed three times with water (5 mL×3), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried. Purification on a silica gel preparative plate (dichloromethane / methanol = 15 / 1, v / v) gave A37 (40 mg, 57%).

[0274] Preparation Example 15

[0275] The synthesis of compound A38 was carried out by the following method:

[0276] Synthesis of intermediate A38-1:

[0277] A3-1 (80 mg, 0.13 mmol) and triethylamine (41 mg, 0.40 mmol) were dissolved in dry acetonitrile (2 mL). After stirring in an ice-water bath for 0.5 h, the reaction mixture was added with tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (29 mg, 0.15 mmol) and potassium iodide (1 mg, 0.010 mmol). The mixture was allowed to react overnight at room temperature under nitrogen. 2 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A38-1 (45 mg, 44%) as a white solid. 1 H NMR (300MHz, DMSO-d6): δ13.41(s,1H),8.36-8.24(m,2H),8.12-8.01(m,2H),7.80(t,J=7.8Hz,1H),7.40(d,J=7.2Hz,1H),3.99-3.92(m, 2H),3.88(s,3H),3.82-3.66(m,7H),3.65-3.42(m,13H),3.27-3.20(m,2H),3.05-2.95(m,2H),2.23-2.01(m,4H),1.36(s,9H).LC-MS:m / z 755.9[M+H] + .

[0278] Synthesis of intermediate A38-2:

[0279] A38-1 (45 mg, 0.060 mmol) was dissolved in dry dichloromethane (1 mL). Trifluoroacetic acid (1 mL) was added in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated, and dichloromethane (2 mL) was added to the residue. The pH of the solution was adjusted to 7-8 with saturated aqueous NaHCO₃ in an ice-water bath. The solution was extracted three times with 15 mL of dichloromethane (5 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure to obtain crude product A38-2 (30 mg, 77%), which was used directly in the next reaction. LC-MS: m / z 656.1 [M+H] + .

[0280] Synthesis of final product A38:

[0281] Intermediate A38-2 (30 mg, 0.050 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (15 mg, 0.050 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (18 mg, 0.14 mmol) was added and stirred at 90°C for 12 hours. Water (2 mL) was added to the mixture, which was then extracted three times with 30 mL of ethyl acetate (10 mL x 3) and washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 10 / 1, v / v) to yield A38 (4 mg, 10%).

[0282] Preparation Example 16

[0283] The synthesis of compound A40 was carried out by the following method:

[0284] Synthesis of intermediate A40-1:

[0285] Dissolve tert-butyl 3-fluoro-3-(hydroxymethyl)azetidine-1-carboxylate (500 mg, 2.4 mmol) in dichloromethane (15 mL). Add DMP (1.2 g, 2.9 mmol) under ice-cooling. Slowly return the mixture to room temperature. The reaction is complete after 2 hours. Add saturated sodium thiosulfate pentahydrate solution (10 mL) to quench the reaction. Add saturated sodium bicarbonate solution (10 mL) and filter through celite. The filtrate is extracted with dichloromethane (15 mL x 2), dried over anhydrous sodium sulfate, filtered, and dried to obtain crude product A40-1 (300 mg) as a white solid. LC-MS: m / z 257.9 [M+MeOH+Na] + .

[0286] Synthesis of intermediate A40-2:

[0287] P-1 (50 mg, 0.19 mmol) and A40-1 (120 mg, 0.59 mmol) were dissolved in DCE (2 mL) and stirred at room temperature for 1 hour. Then, sodium triacetoxyborohydride (100 mg, 0.48 mmol) was reacted at room temperature overnight. After the reaction was completed, saturated sodium bicarbonate solution (10 mL) was added to quench the reaction and extracted with dichloromethane (20 mL × 2). The reaction was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was obtained by silica gel column chromatography (dichloromethane / methanol = 30 / 1) to give a white solid. The crude product was dissolved in dichloromethane (4 mL) and trifluoroacetic acid (2 mL) was added. The reaction was allowed to react at room temperature for 1 hour. After completion of the reaction, the solvent was dried, and saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8. The product was extracted with dichloromethane (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and dried. The product was then slurried with diethyl ether (5 mL), filtered, and dried to obtain A40-2 (30 mg, 26%) as a white solid. LC-MS: m / z 604.9 [M+H] + .

[0288] Synthesis of final product A40:

[0289] A40-2 (30 mg, 0.050 mmol) was dissolved in dimethyl sulfoxide (1.5 mL), and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (16 mg, 0.058 mmol) and DIPEA (32 mg, 0.25 mmol) were added. The reaction was allowed to proceed at 90 ° C overnight. After the reaction was completed, dichloromethane (10 mL) was added for extraction, and the mixture was washed three times with water (5 mL × 3), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried. Purification on a silica gel preparative plate (dichloromethane / methanol = 15 / 1, v / v) gave A40 (22 mg, 51%).

[0290] Preparation Example 17

[0291] The synthesis of compound A41 was carried out by the following method:

[0292] Synthesis of intermediate A41-1:

[0293] (S)-N-Boc-2-hydroxymethylmorpholine (100 mg, 0.46 mmol) was dissolved in dry dichloromethane (2 mL). Trifluoroacetic acid (2 mL) was added in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated, and dichloromethane (2 mL) was added to the residue. The pH of the solution was adjusted to 7-8 with solid NaHCO3 powder in an ice-water bath. The solution was filtered and concentrated under reduced pressure to yield crude product A41-1 (50 mg, 93%), which was used directly in the next reaction.

[0294] Synthesis of intermediate A41-2:

[0295] Intermediate A41-1 (50 mg, 0.43 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (107 mg, 0.39 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (200 mg, 1.6 mmol) was added and stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The mixture was washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A41-2 (110 mg, 76%) as a yellow solid. 1 H NMR (300MHz, DMSO-d6): δ11.09(s,1H),7.71(d,J=8.7Hz,1H),7.35(s,1H),7.27(d,J=8.4Hz,1H),5.13-5.02(m,1H),4.88-4.76(m ,1H),4.00-3.81(m,2H),3.67-3.45(m,3H),2.99-2.82(m,2H),2.75-2.65(m,1H),2.63-2.51(m,4H),2.08-1.96(m,1H).LC-MS:m / z 373.9[M+H] + .

[0296] Synthesis of intermediate A41-3:

[0297] Intermediate A41-2 (30 mg, 0.080 mmol) was dissolved in dry dichloromethane (2 mL). Dess-Martin reagent (41 mg, 0.10 mmol) was added in an ice-water bath, and the reaction was stirred at room temperature for 2 hours. The reaction solution was quenched by adding saturated sodium sulfite solution (1 mL) and saturated sodium bicarbonate solution (2 mL). The solution was extracted three times with 30 mL of dichloromethane (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to obtain A41-3 (20 mg, 67%) as a yellow solid. LC-MS: m / z 403.9 [M+MeOH+H] + .

[0298] Synthesis of final product A41:

[0299] A41-3 (20 mg, 0.050 mmol) and P-1 (25 mg, 0.050 mmol) were dissolved in dry 1,2-dichloroethane (3 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, followed by stirring at room temperature for half an hour. Sodium triacetoxyborohydride (31 mg, 0.15 mmol) was then added to the reaction system, followed by stirring at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A41 (4 mg, 9%).

[0300] Preparation Example 18

[0301] The synthesis of compound A44 was carried out by the following method:

[0302] Synthesis of intermediate A44-1:

[0303] P-1 (100 mg, 0.19 mmol), 2-bromoethanol (238 mg, 1.9 mmol), and potassium carbonate (106 mg, 0.77 mmol) were dissolved in DMF (3 mL) and reacted at 80°C for 6 hours. After completion of the reaction, the mixture was extracted with water (10 mL) and dichloromethane (15 mL × 2). The mixture was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and dried. The residue was slurried with diethyl ether (5 mL), filtered, and dried to obtain A44-1 (80 mg, 73%) as a white solid. LC-MS: m / z 562.0 [M+H] + .

[0304] Synthesis of intermediate A44-2:

[0305] Oxalyl chloride (70 mg, 0.55 mmol) was dissolved in DCM (1 mL) and allowed to stand at -78°C. After 5 minutes, DMSO (86 mg, 1.1 mmol) was added, followed by A44-1 (60 mg, 0.11 mmol) 30 minutes later. After another 30 minutes, triethylamine (172 mg, 1.7 mmol) was added. The reaction was continued for 30 minutes, then brought to room temperature and allowed to react for 1.5 hours. After completion, the reaction was quenched with saturated sodium bicarbonate solution (5 mL) and extracted with dichloromethane (15 mL x 2). The product was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and dried. The product was slurried with diethyl ether (5 mL), filtered, and dried to obtain A44-2 (40 mg, 67%) as a pale yellow solid. LC-MS: m / z 591.9 [M+MeOH+H] + .

[0306] Synthesis of intermediate A44-3:

[0307] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (200 mg, 0.72 mmol) was dissolved in DMSO (5 mL), and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (158 mg, 0.80 mmol) and DIPEA (460 mg, 3.6 mmol) were added and reacted at 90°C overnight. After completion of the reaction, water (10 mL) and dichloromethane (25 mL × 2) were added for extraction, washed with saturated sodium chloride solution (5 mL × 3), dried over anhydrous sodium sulfate, filtered, and dried. The product was slurried with diethyl ether (10 mL), filtered, and dried to obtain a crude yellow solid (270 mg). The crude product was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for two hours. After completion of the reaction, the solvent was dried to obtain a crude yellow solid A44-3 (290 mg). LC-MS: m / z 354.9 [M+H] + .

[0308] Synthesis of final product A44:

[0309] A44-3 (25 mg, 0.054 mmol) was dissolved in DCE (0.5 mL), and triethylamine (18 mg, 0.18 mmol) was added. After 30 minutes, A44-2 (20 mg, 0.036 mmol) was added, and sodium triacetoxyborohydride (23 mg, 0.11 mmol) was added after another 30 minutes. The reaction was allowed to react overnight. After completion of the reaction, saturated sodium bicarbonate solution (3 mL) was added to quench the reaction and extracted with dichloromethane (10 mL × 2). The reaction was washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried. A44 (8 mg, 25%) was obtained after purification on a silica gel preparative plate (dichloromethane / methanol = 15 / 1, v / v).

[0310] Preparation Example 19

[0311] The synthesis of compound A45 was carried out by the following method:

[0312] Synthesis of intermediate A45-1:

[0313] Dissolve 5-hydroxy-2-methoxypyridine (4.8 g, 38 mmol) in concentrated sulfuric acid (20 mL). Slowly add potassium nitrate (4.6 g, 46 mmol) in portions at -10°C. Maintain stirring at low temperature for 2 hours. Pour the reaction solution into ice water, filter, and dry to obtain A45-1 (5.0 g, 77%) as a yellow solid. 1 H NMR (300MHz, CDCl3): δ10.18 (s, 1H), 7.54 (d, J = 9.0Hz, 1H), 7.11 (d, J = 9.0Hz, 1H), 3.98 (s, 3H).

[0314] Synthesis of intermediate A45-2:

[0315] A45-1 (4.0 g, 24 mmol) and ammonium chloride (24 g, 470 mmol) were dissolved in a mixture of tetrahydrofuran and water (80 mL / 30 mL). Zinc powder (12 g, 188 mmol) was slowly added at 50°C and stirred for 5 hours. The reaction mixture was filtered, diluted with saturated aqueous sodium bicarbonate (30 mL), extracted with ethyl acetate (20 mL × 3), dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1, v / v) to obtain A45-2 (2.7 g, 82%) as a black solid. 1 HNMR (300MHz, DMSO-d6): δ8.69(s,1H),6.95-6.70(m,1H),5.87-5.66(m,1H),5.35(s,2H),3.65(s,3H).

[0316] Synthesis of intermediate A45-3:

[0317] A45-2 (1.0 g, 7.1 mmol) was dissolved in pyridine (20 mL), and potassium ethylxanthate (1.7 g, 11 mmol) was added. The mixture was stirred and refluxed at 120°C overnight. The reaction mixture was poured into ice water, acidified with 3N aqueous HCl, filtered, and washed with water to obtain A45-3 (1.0 g, 77%) as a yellow solid. 1 H NMR (300MHz, CDCl3): δ7.49 (d, J = 8.8Hz, 1H), 6.60 (d, J = 9.3Hz, 1H), 3.92 (s, 3H). LC-MS: m / z 183.1 [M+H] + .

[0318] Synthesis of intermediate A45-4:

[0319] A45-3 (1.1 g, 6.0 mmol) was dissolved in ethyl acetate (20 mL), and potassium carbonate (1.6 g, 12 mmol) and iodomethane (1.7 g, 12 mmol) were added. The mixture was stirred at room temperature for 3 hours. The reaction mixture was evaporated to dryness, diluted with water (20 mL), extracted with ethyl acetate (20 mL x 3), dried over anhydrous sodium sulfate, and concentrated to afford A45-4 (1.8 g, 81%) as a yellow solid. 1 H NMR (300MHz, CDCl3): δ7.81 (d, J = 8.4Hz, 1H), 6.82 (d, J = 7.5Hz, 1H), 4.19 (s, 3H), 2.98 (s, 3H).

[0320] Synthesis of intermediate A45-5:

[0321] A45-4 (17 g, 87 mmol) and N-tert-butyloxycarbonylpiperazine (81 g, 434 mmol) were dissolved in tetrahydrofuran (120 mL) and stirred at 90°C overnight. The reaction mixture was concentrated, and the residue was slurried with water, filtered, and dried to obtain A45-5 (20.4 g, 70%) as a gray solid. 1 H NMR (300MHz, DMSO-d6): δ7.71 (d, J = 8.7Hz, 1H), 6.38 (d, J = 8.7Hz, 1H), 3.81 (s, 3H), 3.65-3.55 (m, 4H), 3.52-3.44 (m, 4H), 1.42 (s, 9H). LC-MS: m / z 334.9[M+H] + .

[0322] Synthesis of intermediate A45-6:

[0323] A45-5 (20 g, 60 mmol) was dissolved in acetonitrile (160 mL). N-bromosuccinimide (13 g, 72 mmol) was added portionwise and stirred at room temperature for 2 hours. The reaction mixture was concentrated, and water (120 mL) was added to the residue. The mixture was extracted three times with 900 mL of ethyl acetate (300 mL x 3). The mixture was washed three times with 900 mL of saturated NaCl solution (300 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (petroleum ether / ethyl acetate = 5 / 1, v / v) to yield A45-6 (17 g, 69%) as a brown solid. 1 H NMR (300MHz, DMSO-d6): δ8.13(s,1H),3.88(s,3H),3.67-3.58(m,4H),3.51-3.43(m,4H),1.42(s,9H).LC-MS:m / z412.8[M+H] + .

[0324] Synthesis of intermediate A45-7:

[0325] A45-6 (17 g, 41 mmol), potassium vinyl trifluoroborate (6.1 g, 45 mmol), potassium carbonate (11 g, 82 mmol), and Pd(dppf)Cl2 (1.5 g, 2.1 mmol) were dissolved in a 1,4-dioxane / water mixture (400 mL / 40 mL) and stirred at 100°C overnight under nitrogen. The reaction mixture was concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A45-7 (8.6 g, 58%) as a pale yellow solid. 1H NMR (300MHz, DMSO-d6): δ7.99(s,1H),6.85(dd,J=17.4,11.4Hz,1H),5.78(d,J=17.4Hz,1H),5. 19(d,J=11.4Hz,1H),3.87(s,3H),3.69-3.57(m,4H),3.54-3.43(m,4H),1.43(s,9H).LC-MS:m / z 361.0[M+H] + .

[0326] Synthesis of intermediate A45-8:

[0327] A45-7 (7.0 g, 19 mmol) was dissolved in a dichloromethane / methanol mixture (80 mL / 8 mL) and ozone was introduced at -78°C for 2 hours. The mixture was then quenched by the addition of dimethyl sulfide (8 mL). After being brought to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A45-8 (4.8 g, 68%) as a pale yellow solid. 1 H NMR (300MHz, DMSO-d6): δ10.15(s,1H),7.90(s,1H),3.98(s,3H),3.76-3.62(m,4H),3.55-3.46(m,4H),1.43(s,9H).LC-MS:m / z 362.9[M+M] + .

[0328] Synthesis of intermediate A45-9:

[0329] A45-8 (4.8 g, 13 mmol) was dissolved in tert-butanol (150 mL), and isopentene (9.3 g, 133 mmol) was added at room temperature. Sodium dihydrogen phosphate dihydrate (24 g, 133 mmol) and sodium chlorite (6.0 g, 66 mmol) were each dissolved in 75 mL of water and added sequentially to the reaction mixture. The reaction mixture was stirred at room temperature for 5 hours. The pH of the reaction mixture was adjusted to 3-4 with 1N aqueous hydrochloric acid. The mixture was extracted three times with 900 mL of dichloromethane (300 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure to obtain A45-9 (3.0 g, 60%) as a white solid. 1 H NMR (300MHz, DMSO-d6): δ8.04(s,1H),3.89(s,3H),3.71-3.59(m,4H),3.54-3.42(m,4H),1.43(s,9H).LC-MS:m / z 378.9[M+H] + .

[0330] Synthesis of intermediate A45-10:

[0331] A45-9 (1.7 g, 4.5 mmol) was dissolved in dry N,N-dimethylacetamide (20 mL). N,N-diisopropylethylamine (2.9 g, 22 mmol) and 2-amino-6-(1-methyl-1H-pyrazol-4-yl)pyridine (1.2 g, 6.7 mmol) were added. A 50% T3P solution (15 mL, 22 mmol) in ethyl acetate was added with stirring, and the mixture was stirred at 100°C overnight. The reaction mixture was poured into ice water (20 mL) and extracted three times with 120 mL of ethyl acetate (40 mL x 3). The mixture was washed three times with 120 mL of saturated NaCl solution (40 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A45-10 (1.3 g, 54%) as a brown solid. 1 HNMR (300MHz, DMSO-d6): δ10.39(s,1H),8.32-8.17(m,2H),8.08-7.94(m,2H),7.83-7.75(m,1H),7.40( d,J=4.2Hz,1H),4.16(s,3H),3.90(s,3H),3.74-3.61(m,4H),3.56-3.46(m,4H),1.43(s,9H).LC-MS:m / z 534.9[M+H] + .

[0332] Synthesis of intermediate A45-11:

[0333] A45-10 (1.3 g, 2.4 mmol) was dissolved in dry ethyl acetate (5 mL). 2.8 N hydrochloric acid in ethyl acetate (6 mL) was added in an ice-water bath, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated, and the residue was added with 5 mL of ethyl acetate. The pH of the solution was adjusted to 7-8 with saturated NaHCO₃ solution in an ice-water bath. The solution was extracted three times with 90 mL of ethyl acetate (30 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure to obtain A45-11 (1.0 g, 95%) as a brown solid. 1H NMR (300MHz, DMSO-d6): δ10.38(s,1H),8.28(s,1H),8.21(s,1H),8.09-7.97(m,2H),7.79(t,J=8.1Hz, 1H),7.40(d,J=7.5Hz,1H),4.15(s,3H),3.90(s,3H),3.70-3.55(m,4H),2.91-2.71(m,4H).LC-MS:m / z 434.9[M+H] + .

[0334] Synthesis of intermediate A45-12:

[0335] A45-11 (59 mg, 0.14 mmol) and A6-1 (100 mg, 0.54 mmol) were dissolved in dry 1,2-dichloroethane (3 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (86 mg, 0.41 mmol) was then added to the reaction system, followed by stirring at room temperature for 12 hours. Saturated NH4Cl solution (1 mL) was added to the reaction system to quench the reaction, and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to obtain A45-12 (40 mg, 49%) as a white solid. LC-MS: m / z 603.9 [M+H] + .

[0336] Synthesis of intermediate A45-13:

[0337] A45-12 (40 mg, 0.070 mmol) was dissolved in dry ethyl acetate (1 mL). 2.8 N hydrochloric acid in ethyl acetate (1 mL) was added in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated, and the residue was added with 2 mL of ethyl acetate. The pH was adjusted to 7-8 with saturated aqueous NaHCO₃ in an ice-water bath. The mixture was extracted three times with 15 mL of ethyl acetate (5 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure to obtain crude product A45-13 (30 mg, 90%), which was used directly in the next reaction. 1H NMR (300MHz, DMSO-d6): δ10.37(s,1H),8.33-8.13(m,2H),8.11-7.92(m,2H),7.79(t,J=7.5Hz,1H),7.40(d,J=7.2Hz,1H),4.15 (s,3H),3.89(s,3H),3.79-3.72(m,4H),3.70-3.66(m,4H),3.42-3.35(m,4H),2.89-2.81(m,1H),2.71-2.56(m,2H).LC-MS:m / z 503.9[M+H] + .

[0338] Synthesis of final product A45:

[0339] Intermediate A45-13 (30 mg, 0.060 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (20 mg, 0.070 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (38 mg, 0.30 mmol) was added, and the mixture was stirred at 90°C for 12 hours. Water (3 mL) was added to the mixture, and a yellow solid precipitated. The residue was filtered and purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A45 (20 mg, 44%).

[0340] Preparation Example 20

[0341] The synthesis of compound A46 was carried out by the following method:

[0342] Synthesis of intermediate A46-1:

[0343] A12-1 (85 mg, 0.17 mmol) and triethylamine (51 mg, 0.50 mmol) were dissolved in dry acetonitrile (3 mL). After stirring in an ice bath for 0.5 h, the reaction mixture was added with tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (36 mg, 0.18 mmol) and potassium iodide (3 mg, 0.017 mmol). The mixture was allowed to react overnight at room temperature under nitrogen. 2 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A46-1 (40 mg, 36%) as a white solid. 1H NMR (300MHz, DMSO-d6) δ10.40(s,1H),8.35–8.23(m,2H),8.10–7.99(m,2H),7.81(t,J=7.5Hz,1H),7.41( d,J=7.2Hz,1H),4.16(s,3H),4.12–3.92(m,6H),3.90(s,3H),3.82–3.52(m,12H),1.36(s,9H).LC-MS:m / z 672.9[M+H] + .

[0344] Synthesis of intermediate A46-2:

[0345] A46-1 (40 mg, 0.060 mmol) was dissolved in dry dichloromethane (1 mL). Trifluoroacetic acid (1 mL) was added under ice-cooling conditions, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and 2 mL of dichloromethane was added to the residue. The pH of the solution was adjusted to 7-8 with saturated NaHCO₃ solution under ice-cooling conditions. The solution was concentrated under reduced pressure to obtain crude product A46-2 (20 mg, 59%), which was used directly in the next reaction. LC-MS: m / z 573.0 [M+H] + .

[0346] Synthesis of final product A46:

[0347] Intermediate A46-2 (20 mg, 0.035 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (12 mg, 0.042 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (14 mg, 0.10 mmol) was added, and the mixture was stirred at 90°C for 12 hours. Water (3 mL) was added to the mixture, and a yellow solid precipitated. The residue was filtered and purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A46 (4 mg, 14%).

[0348] Preparation Example 21

[0349] The synthesis of compound A47 was carried out by the following method:

[0350] Synthesis of intermediate A47-1:

[0351] 3-(5-Bromo-1-oxoisoindol-2-yl)piperidine-2,6-dione (500 mg, 1.6 mmol), 1-(tert-butyloxycarbonyl)piperazine (346 mg, 1.9 mmol), and cesium carbonate (1.5 g, 4.7 mmol) were dissolved in dry DMF (10 mL). RuPhos Pd G3 (CAS: 1445085-77-7) (134 mg, 0.16 mmol) was added and reacted at 95°C overnight under nitrogen. After completion of the reaction, the solvent was evaporated and the product was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1, v / v) to obtain A47-1 (180 mg, 27%) as a white solid. LC-MS: m / z 428.9 [M+H] + .

[0352] Synthesis of intermediate A47-2:

[0353] A47-1 (80 mg, 0.19 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added dropwise. The mixture was reacted at room temperature for 1 hour. After the reaction was complete, the solvent was dried to obtain crude A47-2 (100 mg). LC-MS: m / z 329.1 [M+H] + .

[0354] Synthesis of final product A47:

[0355] The crude product of A47-2 (35 mg) was dissolved in DCE (1 mL), and triethylamine (27 mg, 0.27 mmol) was added. After stirring at room temperature for 30 minutes, A44-2 (30 mg, 0.054 mmol) was added. After another 30 minutes, STAB (34 mg, 0.16 mmol) was added. The mixture was reacted at room temperature for 8 hours. After the reaction was completed, saturated sodium bicarbonate solution (2 mL) was added to quench the reaction. The product was extracted with dichloromethane twice (10 mL × 2), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried. A47 (6 mg, 13%) was obtained by silica gel preparation plate (dichloromethane / methanol = 15 / 1, v / v).

[0356] Preparation Example 22

[0357] The synthesis of compound A49 was carried out by the following method:

[0358] Synthesis of final product A49:

[0359] A45-11 (44 mg, 0.10 mmol) and A50-3 (40 mg, 0.11 mmol) were dissolved in dry 1,2-dichloroethane (2 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (65 mg, 0.31 mmol) was then added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A49 (6 mg, 8%).

[0360] Preparation Example 23

[0361] The synthesis of compound A50 was carried out by the following method:

[0362] Synthesis of intermediate A50-1:

[0363] (R)-1-Boc-3-carboxypyrrolidine methyl ester (400 mg, 1.8 mmol) was dissolved in dry dichloromethane (2 mL). Trifluoroacetic acid (2 mL) was added in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and 3 mL of dichloromethane was added to the residue. The pH of the solution was adjusted to 7-8 with solid NaHCO3 powder in an ice-water bath. The solution was filtered and concentrated under reduced pressure to obtain A50-1 (170 mg, 75%) as an oil. 1 H NMR (300MHz, CDCl3): δ4.81-4.62(m,2H),3.75(s,3H),3.64-3.22(m,4H),2.52-2.16(m,2H).

[0364] Synthesis of intermediate A50-2:

[0365] Intermediate A50-1 (170 mg, 1.3 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (280 mg, 1.0 mmol) were dissolved in dry dimethyl sulfoxide (3 mL). N-ethyldiisopropylamine (523 mg, 4.0 mmol) was added and stirred at 95°C for 12 hours. Water (3 mL) was added to the mixture, which was then extracted three times with 30 mL of ethyl acetate (10 mL x 3) and washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A50-2 (320 mg, 82%) as a yellow solid. 1H NMR (300MHz, CDCl3): δ8.01(s,1H),7.67(d,J=8.4Hz,1H),6.96(s,1H),6.70(d,J=8.4Hz,1H),5.00-4.88(m,1H),3.75(s,3H) ,3.72-3.63(m,2H),3.59-3.44(m,2H),3.35-3.22(m,1H),2.96-2.67(m,3H),2.44-2.30(m,2H),2.19-2.08(m,1H).LC-MS:m / z 385.9[M+H] + .

[0366] Synthesis of intermediate A50-3:

[0367] Intermediate A50-2 (130 mg, 0.34 mmol) was dissolved in dry DCM (2 mL). Under nitrogen, a 1 mol / L DIBAL-H solution in tetrahydrofuran (0.74 mL, 0.74 mmol) was added dropwise to the reaction mixture at -78°C. The mixture was then stirred at -78°C for 1 hour. The temperature of the reaction mixture was slowly raised to 0°C, and anhydrous methanol (1 mL) was added to terminate the reaction. Saturated aqueous citric acid (10 mL) was added to the reaction mixture, resulting in the formation of a white solid. The reaction mixture was stirred in an ice-water bath until the white solid disappeared. The mixture was extracted three times with 30 mL of dichloromethane (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A50-3 (60 mg, 50%) as a yellow solid. 1 H NMR (300MHz, CDCl3): δ9.78(s,1H),8.00(s,1H),7.68(d,J=8.4Hz,1H),6.98(s,1H),6.73(d,J=8.1Hz,1H),4.99-4.87(m,1H) ,3.89-3.77(m,1H),3.64-3.39(m,3H),3.35-3.23(m,1H),2.93-2.65(m,3H),2.48-2.27(m,2H),2.18-2.07(m,1H).LC-MS:m / z 387.9[M+MeOH+H] + .

[0368] Synthesis of final product A50:

[0369] P-1 (26 mg, 0.050 mmol) and A50-3 (20 mg, 0.060 mmol) were dissolved in dry 1,2-dichloroethane (2 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (33 mg, 0.15 mmol) was then added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A50 (4 mg, 9%).

[0370] Preparation Example 24

[0371] The synthesis of compound A51 was carried out by the following method:

[0372] Synthesis of final product A51:

[0373] A45-11 (50 mg, 0.12 mmol) and A52-3 (45 mg, 0.13 mmol) were dissolved in dry 1,2-dichloroethane (2 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (73 mg, 0.35 mmol) was then added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A51 (10 mg, 11%).

[0374] Preparation Example 25

[0375] The synthesis of compound A52 was carried out by the following method:

[0376] Synthesis of intermediate A52-1:

[0377] (S)-1-Boc-3-carboxypyrrolidine methyl ester (300 mg, 1.3 mmol) was dissolved in dry dichloromethane (2 mL). Trifluoroacetic acid (2 mL) was added in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and 3 mL of dichloromethane was added to the residue. The pH of the solution was adjusted to 7-8 with solid NaHCO3 powder in an ice-water bath. The solution was filtered and concentrated under reduced pressure to obtain crude oil A52-1 (120 mg, 71%), which was used directly in the next reaction.

[0378] Synthesis of intermediate A52-2:

[0379] Intermediate A52-1 (120 mg, 0.93 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (214 mg, 0.78 mmol) were dissolved in dry dimethyl sulfoxide (3 mL). N-ethyldiisopropylamine (400 mg, 3.10 mmol) was added and stirred at 90°C for 12 hours. Water (3 mL) was added to the mixture, which was then extracted three times with 30 mL of ethyl acetate (10 mL x 3) and washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A52-2 (200 mg, 67%) as a yellow solid. 1 H NMR (300MHz, DMSO-d6): δ11.08(s,1H),7.65(d,J=8.4Hz,1H),6.93(s,1H),6.84(d,J=7.5Hz,1H),5.10-4.99(m,1H),3 .66(s,3H),3.63-3.46(m,4H),2.94-2.80(m,1H),2.64-2.52(m,2H),2.36-2.16(m,2H),2.13-1.86(m,2H).LC-MS:m / z 385.9[M+H] + .

[0380] Synthesis of intermediate A52-3:

[0381] Intermediate A52-2 (160 mg, 0.42 mmol) was dissolved in dry DCM (2 mL). Under nitrogen, a 1 mol / L DIBAL-H solution in tetrahydrofuran (0.91 mL, 0.91 mmol) was added dropwise to the reaction mixture at -78°C. The mixture was then stirred at -78°C for 1 hour. The temperature of the reaction mixture was slowly raised to 0°C, and anhydrous methanol (1 mL) was added to terminate the reaction. Saturated aqueous citric acid (10 mL) was added to the reaction mixture, resulting in the formation of a white solid. The reaction mixture was stirred in an ice-water bath until the white solid disappeared. The mixture was extracted three times with 30 mL of dichloromethane (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A52-3 (95 mg, 64%) as a yellow solid. 1H NMR (300MHz, CDCl3): δ9.78(s,1H),8.00(s,1H),7.68(d,J=8.4Hz,1H),6.98(s,1H),6.73(d,J=8.1Hz,1H),5.01-4.87(m,1H) ,3.90-3.76(m,1H),3.63-3.40(m,3H),3.35-3.23(m,1H),2.94-2.67(m,3H),2.53-2.26(m,2H),2.21-2.06(m,1H).LC-MS:m / z 387.9[M+MeOH+H] + .

[0382] Synthesis of final product A52:

[0383] P-1 (66 mg, 0.13 mmol) and A52-3 (50 mg, 0.14 mmol) were dissolved in dry 1,2-dichloroethane (2 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (81 mg, 0.38 mmol) was then added to the reaction system, followed by stirring at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A52 (13 mg, 12%).

[0384] Preparation Example 26

[0385] The synthesis of compound A53 was carried out by the following method:

[0386] Synthesis of final product A53:

[0387] The crude product of A44-3 (40 mg) was dissolved in DCE (1 mL), and triethylamine (32 mg, 0.32 mmol) was added. After stirring at room temperature for 30 minutes, A13-2 (30 mg, 0.063 mmol) was added. After another 30 minutes, STAB (40 mg, 0.19 mmol) was added. The mixture was reacted at room temperature for 8 hours. After the reaction was completed, saturated sodium bicarbonate solution (2 mL) was added to quench the reaction. The product was extracted with dichloromethane twice (10 mL × 2), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried. A53 (22 mg, 43%) was obtained by silica gel preparation plate (dichloromethane / methanol = 15 / 1, v / v).

[0388] Preparation Example 27

[0389] The synthesis of compound A55 was carried out by the following method:

[0390] Synthesis of final product A55:

[0391] A41-3 (40 mg, 0.11 mmol) and A45-11 (43 mg, 0.10 mmol) were dissolved in dry 1,2-dichloroethane (3 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (62 mg, 0.29 mmol) was then added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A55 (17 mg, 22%).

[0392] Preparation Example 28

[0393] The synthesis of compound A56 was carried out by the following method:

[0394] Synthesis of intermediate A56-1:

[0395] A45-11 (100 mg, 0.23 mmol) and A40-1 (94 mg, 0.46 mmol) were dissolved in dry 1,2-dichloroethane (3 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (147 mg, 0.69 mmol) was then added to the reaction system, followed by stirring at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford intermediate A56-1 (65 mg, 45%) as a gray solid. 1 H NMR (300MHz, DMSO-d6) δ10.39(s,1H),8.37–8.18(m,2H),8.15–7.95(m,2H),7.80(t,J=7.8Hz,1H),7.40(d,J=7.5Hz, 1H),4.16(s,3H),4.09–3.78(m,7H),3.77–3.57(m,4H),2.98–2.79(m,2H),2.78–2.56(m,4H),1.39(s,9H).LC-MS:m / z 621.9[M+H] + .

[0396] Synthesis of intermediate A56-2:

[0397] Intermediate A56-1 (65 mg, 0.10 mmol) was dissolved in dry dichloromethane (1 mL). Trifluoroacetic acid (1 mL) was added under ice-cooling conditions, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated, and the residue was added with 2 mL of dichloromethane. The pH of the solution was adjusted to 7-8 with saturated NaHCO3 solution under ice-cooling conditions. The solution was concentrated under reduced pressure to obtain crude product A56-2 (35 mg, 64%), which was used directly in the next reaction. LC-MS: m / z 521.9 [M+H] + .

[0398] Synthesis of final product A56:

[0399] Intermediate A56-2 (35 mg, 0.067 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (20 mg, 0.074 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (15 mg, 0.12 mmol) was added and the mixture was stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, and a yellow solid precipitated. This solid was extracted three times with 30 mL of ethyl acetate (10 mL x 3) and washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A56 (8 mg, 15%).

[0400] Preparation Example 29

[0401] The synthesis of compound A57 was carried out by the following method:

[0402] Synthesis of intermediate A57-1:

[0403] A45-11 (50 mg, 0.12 mmol) and 1-Boc-piperidine-4-carboxaldehyde (30 mg, 0.14 mmol) were dissolved in DCE (1.5 mL) and stirred at room temperature for 30 minutes. Sodium triacetoxyborohydride (50 mg, 0.24 mmol) was then added and reacted overnight at room temperature. After completion, the reaction was quenched with saturated sodium bicarbonate solution (5 mL) and extracted with dichloromethane (20 mL × 2). The mixture was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and dried. The mixture was purified by silica gel column chromatography (dichloromethane / methanol = 30 / 1, v / v), slurried with diethyl ether (10 mL), filtered, and dried to obtain A57-1 (50 mg, 68%) as a pale yellow solid. LC-MS: m / z 631.9 [M+H] + .

[0404] Synthesis of intermediate A57-2:

[0405] A57-1 (50 mg, 0.079 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. The mixture was allowed to react at room temperature for 1 hour. After the reaction was complete, the solvent was evaporated, and saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8. The mixture was extracted with dichloromethane (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and dried. The mixture was slurried with diethyl ether (10 mL), filtered, and dried to obtain a white solid (25 mg, 60%). LC-MS: m / z 531.9 [M+H] + .

[0406] Synthesis of final product A57:

[0407] A57-2 (25 mg, 0.047 mmol) was dissolved in dimethyl sulfoxide (0.6 mL), and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindole-1,3-dione (14 mg, 0.052 mmol) and DIPEA (31 mg, 0.24 mmol) were added. The reaction was allowed to proceed at 90°C overnight. After completion of the reaction, dichloromethane (15 mL) was added for extraction, and the mixture was washed three times with water (5 mL × 3), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. A57 (13 mg, 35%) was obtained by chromatography on a silica gel plate (dichloromethane / methanol = 15 / 1, v / v).

[0408] Preparation Example 30

[0409] The synthesis of compound A59 was carried out by the following method:

[0410] Synthesis of final product A59:

[0411] The crude product of A47-2 (42 mg) was dissolved in DCE (1 mL), and triethylamine (32 mg, 0.32 mmol) was added. After stirring at room temperature for 30 minutes, A13-2 (30 mg, 0.063 mmol) was added. After another 30 minutes, STAB (40 mg, 0.19 mmol) was added. The mixture was reacted at room temperature for 8 hours. After the reaction was completed, saturated sodium bicarbonate solution (2 mL) was added to quench the reaction. The product was extracted with dichloromethane twice (10 mL × 2), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and dried. A59 (20 mg, 40%) was obtained by silica gel preparation plate (dichloromethane / methanol = 15 / 1, v / v).

[0412] Preparation Example 31

[0413] The synthesis of compound A60 was carried out by the following method:

[0414] Synthesis of final product A60:

[0415] The crude product of A35-4 (50 mg) was dissolved in DCE (2 mL), and A45-11 (50 mg, 0.12 mmol) was added. After stirring at room temperature for 30 minutes, STAB (74 mg, 0.35 mmol) was added thereto and the mixture was allowed to stand overnight. After the reaction was completed, saturated sodium bicarbonate solution (3 mL) was added to quench the reaction. The mixture was extracted twice with dichloromethane (20 mL × 2), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and dried by spin drying. A60 (45 mg, 52%) was obtained by chromatography on a silica gel column (dichloromethane / methanol = 10 / 1, v / v).

[0416] Preparation Example 32

[0417] The synthesis of compound A61 was carried out by the following method:

[0418] Synthesis of intermediate A61-1:

[0419] Intermediate A45-11 (60 mg, 0.14 mmol) and A67-2 (38 mg, 0.21 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (36 mg, 0.28 mmol) was added and stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The mixture was washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A61-1 (60 mg, 70%) as a gray solid. 1 HNMR (300MHz, DMSO-d6) δ10.39(s,1H),8.29(s,1H),8.23(s,1H),8.11–7.94(m,2H),7.80(t,J=7.8Hz,1H),7.41(d,J=7.5Hz,1H),5.71(s,1H ),4.16(s,3H),3.90(s,3H),3.83–3.76(m,2H),3.74–3.59(m,6H),2.76–2.62(m,4H),2.61–2.55(m,2H),1.38(s,9H).LC-MS:m / z619.9[M+H] + .

[0420] Synthesis of intermediate A61-2:

[0421] A61-1 (60 mg, 0.097 mmol) was dissolved in dry dichloromethane (1 mL). Trifluoroacetic acid (1 mL) was added under ice-cooling conditions, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and 2 mL of dichloromethane was added to the residue. The pH of the solution was adjusted to 7-8 with saturated NaHCO3 solution under ice-cooling conditions. The solution was concentrated under reduced pressure to obtain crude product A61-2 (40 mg, 80%), which was used directly in the next reaction. LC-MS: m / z 519.9 [M+H] + .

[0422] Synthesis of final product A61:

[0423] Intermediate A61-2 (40 mg, 0.077 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (26 mg, 0.092 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (20 mg, 0.15 mmol) was added and the mixture was stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, and a yellow solid precipitated. This solid was extracted three times with 30 mL of ethyl acetate (10 mL x 3) and washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A61 (20 mg, 34%).

[0424] Preparation Example 33

[0425] The synthesis of compound A64 was performed by the following method:

[0426] Synthesis of final product A64:

[0427] A65-3 (40 mg, 0.11 mmol) and A45-11 (39 mg, 0.090 mmol) were dissolved in dry 1,2-dichloroethane (3 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (57 mg, 0.27 mmol) was then added to the reaction system, followed by stirring at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A64 (20 mg, 28%).

[0428] Preparation Example 34

[0429] The synthesis of compound A65 was carried out by the following method:

[0430] Synthesis of intermediate A65-1:

[0431] (R)-N-Boc-2-hydroxymethylmorpholine (250 mg, 1.2 mmol) was dissolved in dry dichloromethane (2 mL). Trifluoroacetic acid (2 mL) was added under ice-cooling conditions, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated, and the residue was added with 2 mL of dichloromethane. The pH of the solution was adjusted to 7-8 with solid NaHCO powder under ice-cooling conditions. The solution was filtered and concentrated under reduced pressure to yield crude product A65-1 (120 mg, 89%), which was used directly in the next reaction.

[0432] Synthesis of intermediate A65-2:

[0433] Intermediate A65-1 (120 mg, 1.0 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (236 mg, 0.85 mmol) were dissolved in dry dimethyl sulfoxide (3 mL). N-ethyldiisopropylamine (441 mg, 3.4 mmol) was added and stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The mixture was washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A65-2 (200 mg, 63%). 1 H NMR (300MHz, DMSO-d6) δ11.09(s,1H),7.71(d,J=8.1Hz,1H),7.35(s,1H),7.27(d,J=7.8Hz,1H),5.14–5.03(m,1H),4.84( t,J=4.8Hz,1H),4.01–3.79(m,3H),3.67–3.43(m,4H),2.99–2.82(m,2H),2.75–2.52(m,3H),2.08–1.96(m,1H).LC-MS:m / z 373.9[M+H] + .

[0434] Synthesis of intermediate A65-3:

[0435] Intermediate A65-2 (140 mg, 0.38 mmol) was dissolved in dry dichloromethane (2 mL). Dess-Martin reagent (191 mg, 0.45 mmol) was added under ice-cooling conditions, and the reaction was stirred at room temperature for 3 hours. The reaction solution was quenched by adding saturated sodium sulfite solution (2 mL) and saturated sodium bicarbonate solution (4 mL). The reaction solution was extracted three times with 45 mL of dichloromethane (15 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to obtain A65-3 (95 mg, 68%) as a yellow solid. LC-MS: m / z 403.9 [M+MeOH+H] + .

[0436] Synthesis of final product A65:

[0437] A65-3 (55 mg, 0.15 mmol) and P-1 (70 mg, 0.13 mmol) were dissolved in dry 1,2-dichloroethane (3 mL). One drop of acetic acid and activated molecular sieves (50 mg) were added, and the mixture was stirred at room temperature for half an hour. Sodium triacetoxyborohydride (86 mg, 0.40 mmol) was then added to the reaction system, followed by stirring at room temperature for 12 hours. The reaction was quenched by the addition of saturated NH4Cl solution (1 mL), and the reaction solution was concentrated under reduced pressure. The residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to afford A65 (25 mg, 21%).

[0438] Preparation Example 35

[0439] The synthesis of compound A67 was carried out by the following method:

[0440] Synthesis of intermediate A67-1:

[0441] tert-Butyl 3-methyleneazetidine-1-carboxylate (1.0 g, 5.9 mmol) was dissolved in dry dimethyl sulfoxide (5 mL). N-bromosuccinimide (2.1 g, 12 mmol) and water (213 mg, 12 mmol) were added under ice-cooling conditions, and the reaction was stirred at room temperature overnight. 5 mL of water was added to the mixture, and the mixture was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The mixture was washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 50 / 1, v / v) to afford a colorless oily mixture of isomers of intermediate A67-1 (1.2 g, 76%), which was used directly in the next reaction.

[0442] Synthesis of intermediate A67-2:

[0443] Intermediate A67-1 (300 mg, 1.1 mmol) was dissolved in dry tetrahydrofuran (5 mL). Under nitrogen, 60% sodium hydride (68 mg, 1.7 mmol) was added in an ice bath. The mixture was stirred at room temperature for 5 hours. The reaction was quenched by adding 1 mL of water and extracted three times with 30 mL of ethyl acetate (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 50 / 1, v / v) to yield A67-2 (25 mg, 12%) as a pale yellow oil. 1 H NMR (300MHz, CDCl3) δ4.29–4.15(m,1H),2.85(s,2H),1.45(s,9H).

[0444] Synthesis of intermediate A67-3:

[0445] Intermediate P-1 (58 mg, 0.11 mmol) and A67-2 (25 mg, 0.14 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (29 mg, 0.23 mmol) was added and stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, which was extracted three times with 30 mL of ethyl acetate (10 mL x 3). The mixture was washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A67-3 (45 mg, 57%) as a gray solid. 1 HNMR(300MHz,DMSO-d6)δ13.41(s,1H),8.38–8.16(m,2H),8.11–7.94(m,2H) ,7.79(t,J=7.2Hz,1H),7.39(d,J=6.3Hz,1H),5.68(s,1H),3.88(s,3H),3.8 3–3.74(m,2H),3.73–3.54(m,6H),3.51–3.43(m,2H),3.31–3.19(m,4H),3.0 8–2.96(m,2H),2.73–2.53(m,6H),2.23–1.98(m,4H),1.38(s,9H).LC-MS:m / z 702.9[M+H] + .

[0446] Synthesis of intermediate A67-4:

[0447] Intermediate A67-3 (45 mg, 0.064 mmol) was dissolved in dry dichloromethane (1 mL). Trifluoroacetic acid (1 mL) was added under ice-cooling conditions, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and 2 mL of dichloromethane was added to the residue. The pH of the solution was adjusted to 7-8 with saturated NaHCO3 solution under ice-cooling conditions. The solution was concentrated under reduced pressure to obtain crude product A67-4 (35 mg, 91%), which was used directly in the next reaction. LC-MS: m / z 603.1 [M+H] + .

[0448] Synthesis of final product A67:

[0449] Intermediate A67-4 (35 mg, 0.058 mmol) and 2-(2,6-dioxypiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (19 mg, 0.070 mmol) were dissolved in dry dimethyl sulfoxide (2 mL). N-ethyldiisopropylamine (15 mg, 0.12 mmol) was added and the mixture was stirred at 90°C for 12 hours. 2 mL of water was added to the mixture, and a yellow solid precipitated. This solid was extracted three times with 30 mL of ethyl acetate (10 mL x 3) and washed three times with 30 mL of saturated NaCl solution (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were concentrated under reduced pressure, and the residue was purified on a silica gel column (dichloromethane / methanol = 20 / 1, v / v) to yield A67 (16 mg, 32%).

[0450] Table 1. Structural formula and spectral data of compounds

[0451] Example 1: IRAK4 protein degradation activity test

[0452] 1. Experimental steps:

[0453] The cells used were human acute monocytic leukemia cell line THP-1 (ATCC), sterile incubator temperature 37-38 ° C, pH range 7.2-7.4, 5% carbon dioxide gas ratio. THP-1 cells (7x10 5After 24 hours of treatment with various compound concentrations, proteins were extracted using RIPA lysis buffer and then boiled at 98°C for 10 minutes in protein loading buffer. 20 μL of the sample was electrophoresed on 7.5% SDS-PAGE and transferred to a NC membrane. After blocking the membrane with 5% skim milk (PBS + 0.05% Tween-20) for 1 hour at room temperature, the membrane was incubated with primary antibodies (including anti-IRAK4, 4363S, CST; anti-Actin, A2066, SIGMA) at 4°C overnight, followed by incubation with secondary antibodies (Goat anti-Rabbit, The cells were detected using an Odyssey CLx dual-color infrared laser imaging system. The positive control was Degrader-5 (Cell Chemical Biology 2020, 27, 1500-1509).

[0454] 2. Experimental results:

[0455] Table 2. Test results of compound degradation activity against IRAK4 in THP-1 cells

[0456] At present, there is no unified standard in the industry for limiting the degree of IRAK4 degradation. After preliminary research and analysis by the present disclosure, the protein degrader disclosed herein is considered to have the potential to produce biological effects when it degrades more than 30% of the IRAK4 protein. Due to the unique physiological functions and mechanisms of PROTAC, the degradation ability and biological effects may not necessarily be linear, that is, PROTAC molecules with weaker degradation ability may also produce better biological effects. The degradation activity test results show that the compounds prepared by the present disclosure can effectively degrade IRAK4 protein in cells, among which compounds A3, A4, A5, A6, A8, A13, A35, A37, A38, A40, A41, A44-A47, A49-A53, A55, A60, and A67 have good degradation activity, all of which are better than the positive control Degrader-5 at the same test level.

[0457] Example 2: FLT3 protein degradation activity test

[0458] 1. Experimental steps:

[0459] The cells used were human acute myeloid leukemia cell line MOLM-13 (ATCC), sterile incubator temperature 37-38°C, pH range 7.2-7.4, 5% carbon dioxide gas ratio. MOLM-13 cells (7x10 5After 24 hours of treatment with various compound concentrations, proteins were extracted using RIPA lysis buffer and then boiled at 98°C for 10 minutes in protein loading buffer. 20 μL of the sample was electrophoresed on 7.5% SDS-PAGE and transferred to a NC membrane. After blocking the membrane with 5% skim milk (PBS + 0.05% Tween-20) for 1 hour at room temperature, the membrane was incubated with primary antibodies (including anti-FLT3, ab245116, Abcam; anti-Actin, A2066, SIGMA) at 4°C overnight, followed by incubation with secondary antibodies (Goat anti-Rabbit, Finally, the samples were detected by Odyssey CLx dual-color infrared laser imaging system.

[0460] 2. Experimental results:

[0461] Table 3. Results of the degradation activity test of the compounds on FLT3 in MOLM-13 cells

[0462] At present, there is no unified standard for the degree of FLT3 degradation in the industry. After preliminary research and analysis by the present disclosure, it is believed that the protein degrader disclosed herein is likely to produce a biological effect when it degrades more than 30% of FLT3 protein. Due to the unique physiological functions and mechanisms of PROTAC, the degradation ability and biological effect may not necessarily be linear. The degradation activity test results show that the compounds prepared by the present disclosure can effectively degrade FLT3 protein in cells, among which compounds A3, A5, A6, A8, A9, A13, A35, A37, A38, A40, A44, A46, A47, A50, A52, A53, A55-A57, A59, A64, A65, and A67 have good degradation activity.

[0463] There are few studies on IRAK4 / FLT3 dual-target protein degraders, and whether their biological effects and synergistic mechanisms require the simultaneous degradation of two target proteins to a large extent remains to be determined.

[0464] Example 3: Test of compound inhibition of tumor cell proliferation

[0465] 1. Experimental steps:

[0466] The cells used were human acute myeloid leukemia cell line MOLM-13 (ATCC), in a sterile incubator at 37-38°C, pH 7.2-7.4, and 5% carbon dioxide gas ratio. MOLM-13 cells (1x10 4Cells were plated in 96-well plates (per well) and various concentrations of compounds were added. The cells were then incubated in an incubator. After 48 hours of cell culture, an equal volume of CellCounting-Lite reagent (DD1101-01, Novozymes) was added and mixed with shaking for 5 minutes. The cells were then incubated at room temperature for 10 minutes. Cell proliferation was then measured using a VICTOR Nivo multi-function plate reader.

[0467] 2. Experimental results:

[0468] Table 4. Test results of the compounds' ability to inhibit tumor cell proliferation

[0469] The present disclosure believes that compound IC 50 ≤500nM, indicating that the compound has good activity in inhibiting tumor cell proliferation. The results of tumor cell proliferation activity test showed that compounds A3, A5, A6, A8, A9, A37, A40, and A44 have activity in inhibiting the proliferation of MOLM-13 tumor cells.

Claims

1. A compound having the structure of formula (I) or a pharmaceutically acceptable salt or solvate thereof: in, R 1 Select one of the following fragments: ULM is selected from one of the following segments: L m is a linking fragment, which consists of m identical, partially identical or different L groups and connects the ULM and the piperazine ring through a covalent bond; m is selected from 1, 2, 3, 4 and 5; Each L is independently selected from the following groups: 1) having 1 to 3 independently selected b Substituted C 3-12 Cycloalkylene; 2) having 1 to 3 independently selected from R b Substituted C 6-10 arylene; 3) having 1 to 3 independently selected from R b substituted 4- to 12-membered heterocyclylene; 4) having 1 to 3 independently selected from R b substituted 5- to 12-membered heteroarylene; 5) having 1 to 3 independently selected from R d Substituted C 1-12 Alkylene; 6) having 1 to 3 independently selected from R d Substituted C 2-12 Vinylene; 7) having 1 to 3 independently selected from R d Substituted C 2-12 Ethynylene; 8) having 1 to 6 ethylene glycol or propylene glycol units; and 9)-C(O)-, -C(O)O-, -O-, -N(R c )-, -S-, -S(O)-, -C(S)-, -C(S)O-, -S(O)2-, -S(O)N(R c )-, -S(O)2N(R c )-, -C(O)-N(R c )-, -N(R c )C(O)-N(R c )- and -OC(O)-N(R c )-; If present, R 4 Selected from hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, C 5-12 Heteroaryl, -OR c 、-C(O)R c 、-C(O)OR c 、-C(O)-N(R c )2、-N(R c )2、-N(R c )C(O)-R c 、-N(R c )C(O)OR c 、-N(R c )C(O)N(R c )2. -OC(O)R c 、-OC(O)-N(R c )2. -SR c 、-S(O)R c 、-S(O)2R c and -S(O)2-N(R c )2; If present, each R b are each independently selected from hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, C 5-12 Heteroaryl, -OR c 、-C(O)R c 、-C(O)OR c 、-C(O)-N(R c )2、-N(R c )2、-N(R c )C(O)-R c 、-N(R c )C(O)OR c 、-N(R c )C(O)N(R c )2. -OC(O)R c 、-OC(O)-N(R c )2. -SR c 、-S(O)R c 、-S(O)2R c and -S(O)2-N(R c )2; if present, the C 1-6 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, C 5-12 The heteroaryl group may be substituted with 1 to 3 R d Replacement; if present, any two non-adjacent R b A bridged ring can be formed through carbon atoms; or two R b Together with the carbon atom to which it is attached, it forms a spiro ring; If present, each R c are each independently selected from hydrogen, C 1-6 Alkyl and C 3-6 Cycloalkyl; If present, each R d Each is independently selected from hydrogen, deuterium, halogen, hydroxy, amino, methylamino, dimethylamino, cyano, methyl, deuterated methyl, methoxy and deuterated methoxy.

2. The compound according to claim 1 or a pharmaceutically acceptable salt or solvate thereof, characterized in that: L m is a linking fragment, which consists of m identical, partially identical or different L groups and connects the ULM and the piperazine ring through a covalent bond; m is selected from 1, 2, 3, 4 and 5; Each L is independently selected from the following groups: 1) having 1 to 3 independently selected b Substituted C 3-12 Cycloalkylene; 2) having 1 to 3 independently selected from R b Substituted C 6-10 arylene; 3) having 1 to 3 independently selected from R b substituted 4- to 12-membered heterocyclylene; 4) having 1 to 3 independently selected from R b substituted 5- to 12-membered heteroarylene; 5) having 1 to 6 ethylene glycol or propylene glycol units; and 6)-C(O)-, -C(O)O-, -O-, -N(R c )-, -C(O)-N(R c )-, -N(R c )C(O)-N(R c )- and -OC(O)-N(R c )-; The R b and R c As defined in claim 1.

3. The compound according to claim 1 or a pharmaceutically acceptable salt or solvate thereof, characterized in that: L m is a linking fragment, which consists of m identical, partially identical or different L groups and connects the ULM and the piperazine ring through a covalent bond; m is selected from 1, 2, 3, 4 and 5; Each L is independently selected from the following groups: (1)-C(O)-, -CH2-, -C(D)(H)-, -C(D)2-, -CR c F-, -CF2-, -O-, -N(R c )-, -C(O)-N(R c )-, -N(R c )C(O)-N(R c )- and -OC(O)-N(R c )-; and (2) One of the following fragments: The R c As defined in claim 1.

4. The compound according to claim 3 or a pharmaceutically acceptable salt or solvate thereof, characterized in that: At least one L is selected from one of the following fragments:

5. The compound according to any one of claims 1, 3 and 4, or a pharmaceutically acceptable salt or solvate thereof, characterized in that: L m Select one of the following fragments:

6. The compound according to any one of claims 1, 3, 4 and 5, or a pharmaceutically acceptable salt or solvate thereof, characterized in that: L m Select one of the following fragments:

7. The following compounds or their pharmaceutically acceptable salts or solvates:

8. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt or solvate thereof; Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.

9. A pharmaceutical combination comprising a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt or solvate thereof or a pharmaceutical composition according to claim 8; Preferably, the pharmaceutical combination further comprises at least one additional drug for preventing and / or treating diseases and / or disorders responsive to IRAK4 and / or FLT3.

10. Use of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 8, or a pharmaceutical combination according to claim 9 in the preparation of a medicament for preventing and / or treating a disease and / or condition that is at least partially responsive to IRAK4 and / or FLT3.

11. The use according to claim 10, characterized in that: Such diseases and / or conditions include: Lymphoma, leukemia, myelodysplastic syndrome, endometriosis, atopic dermatitis, hidradenitis suppurativa, psoriasis, systemic lupus erythematosus, multiple sclerosis, or rheumatoid arthritis.

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