Glutarimide-based aromatic heterocyclic compounds and uses thereof

Abstract

The application provides a glutarimide-based aromatic heterocyclic compound and application thereof, and belongs to the technical field of biological medicines.The glutarimide-based aromatic heterocyclic compound can inhibit the activation of an NLRP3 inflammasome complex by targeting degradation of NEK7, and then regulate the secretion of IL-1beta and IL-18, so that the glutarimide-based aromatic heterocyclic compound can be used for preventing and / or treating diseases related to the function of NEK7 protein or diseases mediated by IL-1beta / IL-18.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to an aromatic heterocyclic compound based on glutarimide or its salts, enantiomers, stereoisomers, solvates, deuterated compounds or polymorphs and their applications, particularly in the treatment of diseases associated with NEK7 or the inflammasome NLRP3 complex. Background Technology

[0002] Inflammatory bodies are multi-protein complexes whose activation plays a central role in innate immunity and inflammation. To date, four inflammatory bodies have been described: the NLRP1, NLRC4, NLRP3, and AIM2 complexes. The NLRP3 inflammatory body complex consists of NLRP3, ASC, and Caspase-1. Its activation leads to the activation of Caspase-1, thereby promoting the secretion of IL-1 and IL-18. IL-1β and IL-18 are inflammatory cytokines mediating various autoimmune diseases, myocardial infarction, metabolic syndrome, inflammatory bowel disease, and macrophage activation syndrome in animal disease models. Therefore, targeting the NLRP3 complex could be an effective treatment for IL-1β and IL-18-mediated diseases.

[0003] Currently, the main targeting approach for the NLRP3 complex is to target NLRP3, one of its core proteins. Numerous NLRP3 inhibitors exist, such as MCC950 (Nat Med. 2015, 21(3):248-55) and its sulfonylurea analogs, including MCC7840 (WO2016131098), Compound 102 (WO2020010118), and compound N14 (J Med Chem 2023, 66(18): 12966–12989); as well as non-MCC950 analogs, such as Compound 23 (WO2024064245A1) and Compound II-8 (J Med Chem 2024, 67(12):9869-9895). Among these, inhibitors such as MCC950 were once included in clinical trials, but these trials were terminated due to liver toxicity. In addition, studies have shown that when the NLRP3 protein is acted upon by small molecule inhibitors, the NLRP3 inflammasome is completely inactivated, which eventually leads to a decrease in the body's immunity (Viruses 2023, 15(6),1339). Therefore, direct targeting of the NLRP3 protein may pose safety concerns.

[0004] NEK7, a member of the NIMA-associated kinases (NEKs) family, is a serine / threonine kinase crucial for mitotic entry, cell cycle progression, cell division, and mitotic processes. It is expressed in various tissues, including the brain, heart, lungs, liver, and spleen. Overexpression of NEK7 induces the production of abnormal cells, which are closely linked to tumors such as retinoblastoma, gallbladder cancer, and head and neck cancer. Recent studies have shown that NEK7 is a key protein in the NLRP3 complex. By binding to the leucine-rich repeat series of NLRP3 in a non-kinase-dependent manner, it promotes the formation of the NLRP3-ASC complex, ASC oligomerization, and Caspase-1 activation, ultimately activating the NLRP3 inflammasome complex. Therefore, targeting NEK7 can inhibit the activation of the NLRP3 inflammasome complex without affecting the NLRP3 protein itself, allowing the body to maintain immunity. This approach overcomes the limitations of NLRP3 inhibitors and is considered a safer method.

[0005] Currently, in the development of drugs targeting the NEK7 protein, Halia Therapeutics has developed NEK7 kinase inhibitors, including HT-6153, HT-6184, HT-6258, HT-6283, and HT-6515, to affect the activity of the NLRP3 inflammasome complex for the treatment of inflammatory diseases. However, mechanistically, the activation of the NLRP3 complex partially depends on the non-kinase activity of NEK7, so the efficacy of the inhibitors may be limited. To address the limitations of kinase inhibitors, Captor Therapeutics has developed the NEK7 molecular glue degrader CPT-513, Novartis has developed the NEK7 molecular glue degrader N7-902, and Monte Rosa has developed the NEK7 degrader MRT-8102. These efforts aim to better target the NLRP3 complex by degrading the entire NEK7 protein; however, all of these NEK7 degraders are currently in the preclinical research stage.

[0006] Therefore, developing more novel and selective high-efficiency NEK7 degraders is of great significance for the treatment of diseases mediated by NLRP3 complexes. Summary of the Invention

[0007] To address the shortcomings of the prior art, this invention provides a new class of aromatic heterocyclic compounds based on glutarimide and their applications. These new aromatic heterocyclic compounds based on glutarimide can degrade NEK7 protein with high activity and selectivity, and inhibit the secretion of IL-1β and IL-18. They can be used to treat inflammatory diseases and tumors related to NEK7 protein function.

[0008] To achieve the above objectives, the specific technical solution of the present invention is as follows:

[0009] In a first aspect, the present invention provides an aromatic heterocyclic compound based on glutarimide, having the structure shown in formula (I):

[0010]

[0011] In formula (I): R 1a R 1b R 1c R 1d Each is independently selected from hydrogen, deuterium, halogen, and halogenated C. 1-10 Alkyl, deuterated C 1-10 Alkyl, C 1-10 Alkoxy, deuterated C 1-10 alkoxy or halogenated C 1-10 Alkoxy;

[0012] Y is selected from CH, CD, CF, CMe, or N;

[0013] X is a chemical bond;

[0014] Cy1 is selected from a monocyclic, bicyclic, or tricyclic skeleton having n substituents R4 or nitrogen atom substitutions. Each substituent R4 may be the same or different and is independently selected from H, deuterium, halogen, -CH3, -CF3, alkynyl, -OH, -SH, -NO2, -NH2, -CN, oxo, or C. 1-6 alkyl;

[0015] Cy2 is selected from unsubstituted or pentanehexasaturated bicyclic or pentanehexasaturated bicyclic with m substituents; each substituent may be the same or different and is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -NO2, -CF2H, oxo, aromatic benzyl, aryl, heteroaromatic or C 1-10 alkyl;

[0016] R w Selected from one or more of H, -CO-W, -CH(CH3)-W, -CH2-W, and -SO2-W; W is a substituted or unsubstituted cyclic hydrocarbon group, saturated heterocyclic group, aryl, heteroaryl, naphthyl, biphenyl, etc. , , , , , , Benzenecycloaryl, thiazolyl, or benzoxane;

[0017] T1, T2, T3, and T4 are each independently selected from CH or N;

[0018] R x Selected from H, halogen, -CF3, C 1-7 Alkyl and R y C with substituent group 1-7 Alkyl or aryl;

[0019] R y Selected from H, F, Cl, -OCH3, -CN, -CF2H, -OCF3, -OPh, -NHPh, -OCH2Ph, -CH3, or -CF3.

[0020] Furthermore, the ring Cy1 is selected from, but not limited to, single-ring, double-ring, or triple-ring skeletons with the following structural forms:

[0021] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ;

[0022] Among them, R 2a R 3a Selected from H or C 1-6 Alkyl; R 2b Selected from F or CH3; R 2c Selected from H or C 1-6 Alkyl; R 4a R 4b Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -OCH3, -OCF3, -OCF2H, alkynyl, or C. 1-6 Alkyl; R 3a Selected from H or C 1-6Alkyl group; the symbol * indicates the junction of the ring Cy1 and X, and the symbol ** indicates the junction of the ring Cy1 and Cy2.

[0023] Furthermore, the ring Cy2 is selected from, but not limited to, pentanehexasaturated bicyclic or pentanehexasaturated bicyclic structures:

[0024] , , , , , , , , , , , or ;

[0025] Among them, R 5a Selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, aromatic benzyl, aryl, or C. 1-6 Alkyl; R 5b R 5c R 5d R 5e Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -OCF3, -OCH3, -CF2H, -OCF2H, or C. 1-10 Alkyl; Z and R 5f The dashed line between them indicates that there is R. 5f Replacement or no R 5f Replacement, when there is R 5f When replacing, Z is N, R 5f Selected from H, -CH3, -CF3, -CH2CF3, aromatic benzyl, aromatic group, heteroaromatic group, or C. 1-10 Alkyl; when R is absent 5f When replacing, Z is O or S; R 6a R 6b R 6c R 6d R 6e Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl; R 7a R 7b R 7c R 7dEach is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl; R 8a R 8b R 8c Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl group; the symbol * indicates the junction of the ring Cy2 and the ring Cy1, and the symbol ** indicates the junction of the ring Cy2 and the group R. w The connection point.

[0026] Furthermore, the halogen includes fluorine, chlorine, bromine, or iodine; the alkyl group includes straight-chain alkyl, branched-chain alkyl, or cycloalkyl;

[0027] The aryl group includes phenyl or fused-ring aryl; the heteroaryl group includes pyrrole, thiophene, furanyl, thiazolyl, pyrazolyl, imidazolyl, pyridinyl, pyrazinyl, oxazolyl, or pyrimidinyl; the cyclic hydrocarbon group is a cycloalkanes having 3 to 7 carbon atoms; the saturated heterocyclic group is a saturated five- to seven-membered ring group having 1 or 2 heteroatoms, and the heteroatoms are one or more of O, N, and S.

[0028] Furthermore, the saturated heterocyclic group is an epoxypentyl group, a cyclothiopentyl group, a p-thionitrohexacycloyl group, or a p-oxynitrohexacycloyl group.

[0029] Further, the fused-ring aryl group includes fused-ring alkylphenyl, fused-ring heteroaryl, fused-ring heterophenyl, fused-ring heteroaryl, fused-ring naphthyl, fused-ring naphthyl, fused-ring anthraquinone, or fused-ring anthraquinone.

[0030] Further, the fused-ring aryl group is benzopyrazinyl, naphthyl, pyridylpyrrole, benzimidazolyl, benzo-p-oxohexaneyl or imidazo-p-nitrohexaneyl.

[0031] Furthermore, the aromatic heterocyclic compound based on glutarimide is selected from the following compounds:

[0032]

[0033]

[0034]

[0035]

[0036]

[0037]

[0038]

[0039] In a second aspect, the present invention provides a pharmaceutical composition comprising an effective amount of one or more of the glutarimide-based aromatic heterocyclic compound or its pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, deuterated compound, and polymorph.

[0040] Thirdly, the present invention provides the use of the glutarimide-based aromatic heterocyclic compound in the preparation of medicaments for the prevention and / or treatment of diseases related to the NEK7 protein.

[0041] Fourthly, the present invention provides the use of the glutarimide-based aromatic heterocyclic compound in the preparation of a medicament for the prevention and / or treatment of diseases related to the NLRP3 inflammasome.

[0042] Fifthly, the present invention provides the use of the glutarimide-based aromatic heterocyclic compound in the preparation of medicaments for the prevention and / or treatment of IL-1β and / or IL-18-mediated related diseases.

[0043] Compared with the prior art, the advantages of the present invention are:

[0044] The aromatic heterocyclic compounds based on glutarimide of the present invention can inhibit the activation of the NLRP3 inflammasome complex by targeting and degrading NEK7, thereby regulating the secretion of IL-1β and IL-18, and thus can be used to prevent and / or treat diseases related to NEK7 protein function or IL-1β / IL-18 mediated diseases, including inflammation, tumors, etc. Attached Figure Description

[0045] Figure 1 Results of experiments showing that LC-06-014 promotes the degradation of NEK7 protein in the Molt4-NEK7-N-HiBiT cell line;

[0046] Figure 2 The release level of IL-1β in hPBMC cells after treatment with MCC950 and LC-06-014;

[0047] Figure 3 The release level of IL-18 in hPBMC cells after treatment with MCC950 and LC-06-014. Detailed Implementation

[0048] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0049] This invention provides an aromatic heterocyclic compound based on glutarimide, having the structure shown in formula (I):

[0050]

[0051] In formula (I): R 1a R 1b R 1c R 1d Each is independently selected from hydrogen, deuterium, halogen, and halogenated carbon. 1-10 Alkyl, deuterated C 1-10 Alkyl, C 1-10 Alkoxy, deuterated C 1-10 alkoxy or halogenated C 1-10 Alkoxy;

[0052] Y is selected from CH, CD, CF, CMe, or N;

[0053] X is a chemical bond;

[0054] Cy1 is selected from a monocyclic, bicyclic, or tricyclic skeleton having n substituents R4 or nitrogen atom substitutions. Each substituent R4 may be the same or different and is independently selected from H, deuterium, halogen, -CH3, -CF3, alkynyl, -OH, -SH, -NO2, -NH2, -CN, oxo, or C. 1-6 alkyl;

[0055] Cy2 is selected from unsubstituted or pentanehexasaturated bicyclic or pentanehexasaturated bicyclic with m substituents; each substituent may be the same or different and is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -NO2, -CF2H, oxo, aromatic benzyl, aryl, heteroaromatic or C 1-10 alkyl;

[0056] R w Selected from one or more of H, -CO-W, -CH(CH3)-W, -CH2-W, and -SO2-W; W is a substituted or unsubstituted cyclic hydrocarbon group, saturated heterocyclic group, aryl, heteroaryl, naphthyl, biphenyl, etc. , , , , , , Benzenecycloaryl, thiazolyl, or benzoxane;

[0057] T1, T2, T3, and T4 are each independently selected from CH or N;

[0058] R x Selected from H, halogen, -CF3, C 1-7 Alkyl and R y C with substituent group 1-7 Alkyl or aryl;

[0059] R y Selected from H, F, Cl, -OCH3, -CN, -CF2H, -OCF3, -OPh, -NHPh, -OCH2Ph, -CH3, or -CF3.

[0060] In some examples, the ring Cy1 is selected from, but not limited to, single-ring, double-ring, or triple-ring skeletons with the following structures:

[0061] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ;

[0062] Among them, R 2a R 3a Selected from H or C 1-6 Alkyl; R 2b Selected from F or CH3; R 2c Selected from H or C 1-6 Alkyl; R 4a R 4b Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -OCH3, -OCF3, -OCF2H, alkynyl, or C. 1-6Alkyl; R 3a Selected from H or C 1-6 Alkyl group; the symbol * indicates the junction of the ring Cy1 and X, and the symbol ** indicates the junction of the ring Cy1 and Cy2.

[0063] In some examples, the ring Cy2 is selected from, but not limited to, pentahexasaturated bicyclic or pentahexasaturated bicyclic structures:

[0064] , , , , , , , , , , , or ;

[0065] Among them, R 5a Selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, aromatic benzyl, aryl, or C. 1-6 Alkyl; R 5b R 5c R 5d R 5e Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -OCF3, -OCH3, -CF2H, -OCF2H, or C. 1-10 Alkyl; Z and R 5f The dashed line between them indicates that there is R. 5f Replacement or no R 5f Replacement, when there is R 5f When replacing, Z is N, R 5f Selected from H, -CH3, -CF3, -CH2CF3, aromatic benzyl, aromatic group, heteroaromatic group, or C. 1-10 Alkyl; when R is absent 5f When replacing, Z is O or S; R 6a R 6b R 6c R 6d R 6e Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl; R 7a R 7b R 7c R 7dEach is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl; R 8a R 8b R 8c Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl group; the symbol * indicates the junction of the ring Cy2 and the ring Cy1, and the symbol ** indicates the junction of the ring Cy2 and the group R. w The connection point.

[0066] In some examples, the halogen includes fluorine, chlorine, bromine, or iodine; the alkyl group includes straight-chain alkyl, branched alkyl, or cycloalkyl; the aryl group includes phenyl or fused-ring aryl; the heteroaryl group includes pyrrole, thiophene, furanyl, thiazolyl, pyrazolyl, imidazolyl, pyridinyl, pyrazinyl, oxazolyl, or pyrimidinyl; the cycloalkyl group is a cycloalkanes having 3 to 7 carbon atoms; the saturated heterocyclic group is a saturated five- to seven-membered ring group having 1 or 2 heteroatoms, wherein the heteroatoms are one or more of O, N, and S.

[0067] In some examples, the saturated heterocyclic group is epoxypentyl, cyclothiopentyl, p-thionitrohexacycloyl, or p-oxynitrohexacycloyl. The fused-ring aryl group includes fused-ring alkylphenyl, fused-ring heteroaryl, fused-ring heterophenyl, fused-ring heteroaryl, fused-ring naphthyl, fused-ring naphthyl, fused-ring anthracene, or fused-ring anthracene.

[0068] In some examples, the fused-ring aryl group is benzopyrazinyl, naphthyl, pyridylpyrrole, benzimidazolyl, benzo-p-oxohexaneyl or imidazo-p-nitrohexaneyl.

[0069] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0070] Example 1: Synthesis of (S)-3-(1-oxo-5-(7-((2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione (LC-04-047)

[0071]

[0072] Step 1: Methyl 4-bromo-2-bromomethylbenzoate (1.3 eq) and (S)-4,5-diamino-5-oxovalerate tert-butyl hydrochloride (1 eq) were dissolved in acetonitrile, and then N,N-diisopropylethylamine (3 eq) was added. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain LC-01-151 (14.52 g, yield 87.25%) as a white solid. LC-MS (ESI) C 17 H 22 BrN2O4 + [M+H] + Calculated values: 397.08 and 399.07; measured values: 397.11 and 399.12.

[0073] Step 2: Dissolve LC-01-151 (1 eq) and pinacol diborate (1.5 eq) in 1,4-dioxane, and add potassium acetate (3 eq). Purge the solution three times with nitrogen, add Pd(dppf)Cl2 (0.03 eq), and purge again three times with nitrogen. Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to obtain a white solid LC-01-156 (14.83 g, yield 89.97%). LC-MS (ESI)C 23 H 34 BN2O6 + [M+H] + Calculated value: 445.25; Measured value: 445.30.

[0074] Step 3: LC-01-156 (1 eq) and 3-bromo-5,6-dihydroimidazolo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) were dissolved in 1,4-dioxane, and 2 M sodium carbonate solution (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(PPh3)4 (0.04 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid, LC-03-038 (3.00 g, yield 80.96%). LC-MS (ESI) C 28 H 38 N5O6 + [M+H] + Calculated value: 540.28; Measured value: 540.30.

[0075] Step 4: Dissolve LC-03-038 (1 eq) in acetonitrile, add benzenesulfonic acid (3 eq), heat the reaction solution to 100℃, and stir for 12 h. Detect the completion of the reaction using LCMS, cool the reaction solution to room temperature, concentrate under reduced pressure to remove the solvent, add ethyl acetate and petroleum ether to the residue, slurry, and filter to obtain a white solid LC-03-041 (2.00 g, yield 94.15%). 1 H NMR (400 MHz, DMSO- d 6) δ 11.04 (s, 1H), 7.87 (d, J = 8.0 Hz, 1H), 7.80 (s,1H), 7.68 (d, J = 8.0 Hz, 1H), 7.63 (s, 1H), 5.16 (dd, J = 12.0, 4.0 Hz, 1H), 4.61 (s, 2H), 4.53 (d, J = 16.0 Hz, 1H), 4.41 (d, J = 16.0 Hz, 1H), 4.35 (t, J = 4.0 Hz, 2H), 3.66 (t, J= 4.0 Hz, 2H), 2.98-2.89 (m, 1H), 2.64-2.59 (m,1H), 2.45-2.38 (m, 1H), 2.04-2.01 (m, 1H). LC-MS (ESI) C 19 H 20 N5O3 + [M+H] + Calculated value: 366.16; Measured value: 366.20.

[0076] Step 5: Synthesis of (S)-3-(1-oxo-5-(7-((2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidine-2,6-dione (LC-03-047)

[0077] LC-03-041 (1 eq) and 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) were dissolved in N,N-dimethylformimide, and a catalytic amount of acetic acid was added. The reaction mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium borohydride acetate (3 eq), and stirring was continued for 12 h. The reaction was detected by LCMS to indicate completion, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel plate to obtain LC-03-047 (32.00 mg, yield 43.99%) as a white solid. LCMS (ESI) C 32 H 34 N5O3 + [M+H] + Calculated value: 536.66; Measured value: 536.70.

[0078] Example 2: Synthesis of (S)-3-(1-oxo-5-(7-((3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)isoindol-2-yl)piperidine-2,6-dione (LC-04-048)

[0079] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 34 N5O3 +[M+H] + Calculated value: 536.66; Measured value: 536.70. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J =7.9 Hz, 1H), 7.70 (s, 1H), 7.60 (d, J = 7.9 Hz, 1H), 7.34 (t, J = 7.5 Hz, 2H), 7.24 (t, J = 7.3 Hz, 1H), 7.18 – 7.09 (m, 3H), 5.13 (dd, J = 13.3, 5.0Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 3.99 (t, J =5.0 Hz, 2H), 3.46 (s, 2H), 3.01 (s, 2H), 2.95 – 2.89 (m, 1H), 2.62 (dd, J =16.7, 11.6 Hz, 3H), 2.45 – 2.39 (m, 1H), 2.26 (s, 2H), 2.22 (s, 2H), 2.04 –1.99 (m, 1H), 1.69 (s, 4H).

[0080] Example 3: Synthesis of (S)-3-(5-(7-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-04-049)

[0081] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI)C 34 H 37 ClN5O3 + [M+H] + Calculated value: 599.15; Measured value: 599.20. 1H NMR (600 MHz, DMSO) δ 11.00(s, 1H), 7.76 (d, J = 7.9 Hz, 1H), 7.72 (s, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.39 (d, J = 8.3 Hz, 2H), 7.21 – 7.11 (m, 3H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 (d, J = 17.3 Hz, 1H), 4.36 (d, J = 17.3 Hz, 1H), 4.01 (t, J = 4.9Hz, 2H), 3.49 (s, 2H), 2.96 (d, J = 7.8 Hz, 2H), 2.94 – 2.87 (m, 1H), 2.62(dd, J = 17.4, 12.0 Hz, 3H), 2.44 – 2.38 (m, 1H), 2.25 (s, 2H), 2.01 (d, J =11.0 Hz, 3H), 1.45 (t, J = 6.3 Hz, 2H), 0.98 (s, 6H).

[0082] Example 4: Synthesis of (S)-3-(5-(7-((2',3'-dichloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-04-157)

[0083] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2',3'-dichloro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 C l2 N5O3 + [M+H] + Calculated value: 600.51; Measured value: 600.50. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J=7.9 Hz, 1H), 7.71 (s, 1H), 7.69 – 7.64 (m, 2H), 7.61 (d, J = 8.0 Hz, 1H),7.47 – 7.39 (m, 3H), 7.34 (dd, J = 7.7, 1.1 Hz, 1H), 7.20 (d, J = 7.5 Hz,1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.36 (d, J = 17.3 Hz, 1H), 4.02 – 3.94 (m, 2H), 3.55 – 3.41 (m, 4H), 2.95 – 2.89 (m, 1H), 2.73 – 2.65 (m, 2H), 2.63 – 2.59 (m, 1H), 2.44 – 2.38(m, 1H), 2.01 (ddd, J = 10.2, 5.1, 3.1 Hz, 1H).

[0084] Example 5: Synthesis of (S)-3-(5-(7-((2',4'-dichloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-04-160)

[0085] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2',4'-dichloro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 Cl2N5O3 + [M+H] + Calculated value: 600.51; Measured value: 600.50. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.74 (dd, J =8.0, 5.0 Hz, 2H), 7.72 (s, 1H), 7.65 (d, J = 7.6 Hz, 1H), 7.62 (d, J= 7.9Hz, 1H), 7.50 (dd, J = 8.2, 2.1 Hz, 1H), 7.47 – 7.44 (m, 1H), 7.39 (t, J =7.8 Hz, 2H), 7.18 (d, J = 7.5 Hz, 1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.1Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.00 (t, J =5.3 Hz, 2H), 3.49 (q, J = 13.5 Hz, 4H), 2.95 – 2.89 (m, 1H), 2.69 (ddd, J =23.2, 11.5, 5.7 Hz, 2H), 2.62 – 2.59 (m, 1H), 2.44 – 2.39 (m, 1H), 2.03 –1.98 (m, 1H).

[0086] Example 6: Synthesis of (S)-3-(5-(7-((2'-chloro-3'-methoxy-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-04-165)

[0087] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-3'-methoxy-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 33 H 30 ClN5O4 + [M+H] + Calculated value: 597.09; Measured value: 597.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.72 (s, 1H), 7.66 (d, J = 7.6 Hz, 1H), 7.61 (d, J= 7.9 Hz,1H), 7.45 – 7.41 (m, 1H), 7.38 – 7.34 (m, 2H), 7.18 (dd, J = 8.3, 0.9 Hz,1H), 7.16 – 7.11 (m, 2H), 6.91 (d, J = 7.2 Hz, 1H), 5.13 (dd, J = 13.3, 5.1Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.01 (t, J =5.4 Hz, 2H), 3.89 (s, 3H), 3.54 – 3.39 (m, 4H), 2.95 – 2.89 (m, 1H), 2.75 –2.64 (m, 2H), 2.62 – 2.58 (m, 1H), 2.44 – 2.38 (m, 1H), 2.03 – 1.97 (m, 1H).

[0088] Example 7: Synthesis of (S)-3-(5-(7-((2'-chloro-3'-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-04-170)

[0089] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-3'-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J =7.9 Hz, 1H), 7.71 (s, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.61 (d, J = 7.9 Hz,1H), 7.49 – 7.43 (m, 3H), 7.40 (t,J = 7.4 Hz, 1H), 7.23 (dd, J = 9.0, 4.3Hz, 1H), 7.20 (d, J = 7.5 Hz, 1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 3.99 (t, J = 5.2Hz, 2H), 3.56 – 3.43 (m, 4H), 2.95 – 2.86 (m, 1H), 2.76 – 2.65 (m, 2H), 2.60(d, J = 16.5 Hz, 1H), 2.42 (dt, J = 13.5, 8.9 Hz, 1H), 2.01 (dt, J = 10.0, 3.9 Hz, 1H).

[0090] Example 8: Synthesis of (S)-2-chloro-2'-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-[1,1'-biphenyl]-3-cyano (LC-04-172)

[0091] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-chloro-2'-formyl-[1,1'-biphenyl]-3-cyano. LCMS (ESI) C 33 H 28 ClN6O3 + [M+H] + Calculated value: 592.08; Measured value: 592.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 8.01 (d, J = 7.7 Hz, 1H), 7.76 – 7.70 (m, 3H), 7.64 (d, J = 7.6 Hz, 1H), 7.61 (t, J =7.5 Hz, 2H), 7.48 (d,J = 7.6 Hz, 1H), 7.43 (t, J = 7.5 Hz, 1H), 7.23 (d, J =7.5 Hz, 1H), 7.14 (s, 1H), 5.13 (dd, J = 13.3, 5.0 Hz, 1H), 4.47 (d, J = 17.4Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 3.99 – 3.91 (m, 2H), 3.54 – 3.42 (m, 4H), 2.94 – 2.89 (m, 1H), 2.72 – 2.64 (m, 2H), 2.61 (d, J = 15.6 Hz, 1H), 2.43 –2.38 (m, 1H), 2.04 – 2.00 (m, 1H).

[0092] Example 9: Synthesis of (S)-3-(5-(7-((2'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-023)

[0093] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-4'-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J =7.9 Hz, 1H), 7.72 (s, 1H), 7.63 (dd, J = 14.3, 7.8 Hz, 2H), 7.56 (dd, J =8.8, 2.1 Hz, 1H), 7.42 (ddd, J = 20.6, 14.5, 7.4 Hz, 3H), 7.30 (td, J= 8.3, 2.1 Hz, 1H), 7.18 (d, J = 7.5 Hz, 1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.0Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.01 (t, J =4.5 Hz, 2H), 3.48 (dd, J = 53.0, 10.6 Hz, 4H), 2.95 – 2.88 (m, 1H), 2.69 (dt, J = 12.2, 9.1 Hz, 2H), 2.60 (d, J = 15.8 Hz, 1H), 2.41 (dd, J = 13.3, 9.2 Hz, 1H), 2.01 (dd, J = 10.5, 5.4 Hz, 1H).

[0094] Example 10: Synthesis of (S)-3-(5-(7-((2'-chloro-4'-methoxy-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-024)

[0095] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-4'-methoxy-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 33 H 31 ClN5O4 + [M+H] + Calculated value: 597.09; Measured value: 597.10. 1 H NMR (600 MHz, DMSO) δ 10.99 (s, 1H), 7.79 –7.70 (m, 2H), 7.63 (dd, J = 14.2, 7.8 Hz, 2H), 7.42 (t, J = 7.4 Hz, 1H), 7.35(t, J = 7.4 Hz, 1H), 7.26 (d,J = 8.5 Hz, 1H), 7.19 – 7.10 (m, 3H), 6.99 (dd, J = 8.5, 2.3 Hz, 1H), 5.13 (dd, J = 13.3, 5.0 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.02 (s, 2H), 3.82 (s, 3H), 3.56 – 3.41 (m,4H), 2.95 – 2.87 (m, 1H), 2.78 – 2.64 (m, 2H), 2.60 (d, J = 15.6 Hz, 1H), 2.44 – 2.37 (m, 1H), 2.01 (dd, J = 6.2, 4.1 Hz, 1H).

[0096] Example 11: Synthesis of (S)-2-chloro-2'-((3-(2-(2-(2-6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazolium[1,2-a]pyrazin-7(8H)-yl)methyl)-[1,1'-biphenyl]-4-cyano (LC-05-025)

[0097] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-chloro-2'-formyl-[1,1'-biphenyl]-4-cyano. LCMS (ESI) C 33 H 28 ClN6O3 + [M+H] + Calculated value: 592.08; Measured value: 592.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 8.18 (s,1H), 7.92 – 7.86 (m, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.71 (s, 1H), 7.65 – 7.58(m, 3H), 7.48 (t, J = 7.4 Hz, 1H), 7.42 (t, J = 7.4 Hz, 1H), 7.20 (d, J= 7.4Hz, 1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.0 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.36 (d, J = 17.3 Hz, 1H), 3.96 (t, J = 4.6 Hz, 2H), 3.51 (dd, J = 43.6,17.8 Hz, 4H), 2.95 – 2.88 (m, 1H), 2.73 – 2.65 (m, 2H), 2.61 (d, J = 16.1 Hz,1H), 2.44 – 2.38 (m, 1H), 2.03 – 1.98 (m, 1H).

[0098] Example 12: Synthesis of (S)-3-(5-(7-((2'-chloro-5'-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-026)

[0099] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-5'-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J =7.9 Hz, 1H), 7.71 (s, 1H), 7.66 (d, J = 7.6 Hz, 1H), 7.61 (dd, J = 8.3, 5.3Hz, 2H), 7.49 – 7.44 (m, 1H), 7.39 (t, J = 7.5 Hz, 1H), 7.33 – 7.26 (m, 2H), 7.19 (d, J= 7.5 Hz, 1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47(d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.08 – 3.93 (m, 2H), 3.52(dt, J = 27.0, 14.1 Hz, 4H), 2.92 (ddd, J = 17.5, 13.7, 5.4 Hz, 1H), 2.77 –2.65 (m, 2H), 2.63 – 2.58 (m, 1H), 2.44 – 2.38 (m, 1H), 2.01 (ddd, J = 10.1, 5.2, 3.1 Hz, 1H).

[0100] Example 13: Synthesis of (S)-3-(5-(7-((2'-chloro-5'-methoxy-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-027)

[0101] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-5'-methoxy-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 33 H 31 ClN5O4 + [M+H] + Calculated value: 597.09; Measured value: 597.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.71 (s, 1H), 7.65 (d, J = 7.6 Hz, 1H), 7.61 (d, J = 8.0 Hz,1H), 7.47 – 7.42 (m, 2H), 7.37 (t, J = 7.2 Hz, 1H), 7.17 (d, J= 7.0 Hz, 1H),7.15 (s, 1H), 6.99 (dd, J = 8.9, 3.0 Hz, 1H), 6.89 (d, J = 3.0 Hz, 1H), 5.13(dd, J = 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz,1H), 4.07 – 3.95 (m, 2H), 3.73 (s, 3H), 3.57 – 3.45 (m, 4H), 2.97 – 2.87 (m,1H), 2.71 (ddd, J = 22.4, 11.8, 5.9 Hz, 2H), 2.63 – 2.57 (m, 1H), 2.45 – 2.38(m, 1H), 2.01 (dt, J = 4.9, 3.9 Hz, 1H).

[0102] Example 14: Synthesis of (S)-6-chloro-2'-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-[1,1'-biphenyl]-3-cyano (LC-05-028)

[0103] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 6-chloro-2'-formyl-[1,1'-biphenyl]-3-cyano. LCMS (ESI) C 33 H 28 ClN6O3 + [M+H] + Calculated value: 592.08; Measured value: 592.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.92 – 7.87(m, 2H), 7.79 (d, J = 8.2 Hz, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.70 (s, 1H), 7.63 (d, J = 7.6 Hz, 1H), 7.61 (d, J= 7.9 Hz, 1H), 7.48 (t, J = 7.4 Hz, 1H), 7.42 (t, J = 7.4 Hz, 1H), 7.22 (d, J = 7.4 Hz, 1H), 7.14 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H),3.99 – 3.93 (m, 2H), 3.56 – 3.44 (m, 4H), 2.95 – 2.89 (m, 1H), 2.71 – 2.65(m, 2H), 2.63 – 2.59 (m, 1H), 2.44 – 2.38 (m, 1H), 2.02 (dd, J = 8.9, 3.5 Hz, 1H).

[0104] Example 15: Synthesis of (S)-3-(5-(7-((2',5'-dichloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-029)

[0105] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2',5'-dichloro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 Cl2N5O3 + [M+H] + Calculated value: 600.51; Measured value: 600.50. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J =7.9 Hz, 1H), 7.71 (s, 1H), 7.64 (d, J = 7.6 Hz, 1H), 7.60 (t, J = 8.2 Hz, 2H), 7.49 (dd, J= 8.6, 2.6 Hz, 1H), 7.48 – 7.44 (m, 2H), 7.39 (dd, J = 10.8,4.2 Hz, 1H), 7.22 – 7.18 (m, 1H), 7.15 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.36 (d, J = 17.3 Hz, 1H), 4.04 – 3.95 (m,2H), 3.55 – 3.46 (m, 4H), 2.95 – 2.88 (m, 1H), 2.70 (qd, J = 12.0, 6.1 Hz,2H), 2.63 – 2.58 (m, 1H), 2.45 – 2.37 (m, 1H), 2.01 (ddd, J = 10.1, 5.2, 3.1Hz, 1H).

[0106] Example 16: Synthesis of (S)-3-(5-(7-(2-(6-chlorobenzo[d][1,3]dioxo-5-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-030)

[0107] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(6-chlorobenzo[d][1,3]dioxol-5-yl)benzaldehyde. LCMS (ESI) C 33 H 29 ClN5O5 + [M+H] + Calculated value: 611.08; Measured value: 611.10. 1 H NMR (600 MHz, DMSO) δ 10.99 (s, 1H), 7.77 –7.71 (m, 2H), 7.63 (dd, J = 10.6, 8.3 Hz, 2H), 7.41 (td, J = 7.6, 1.1 Hz,1H), 7.37 – 7.32 (m, 1H), 7.18 (s, 1H), 7.15 (s, 1H), 7.14 (dd,J = 7.6, 0.8Hz, 1H), 6.92 (s, 1H), 6.11 (d, J = 7.5 Hz, 2H), 5.12 (dd, J = 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.03 (ddt, J =17.5, 11.5, 5.9 Hz, 2H), 3.56 – 3.48 (m, 4H), 2.91 (ddd, J = 17.5, 13.7, 5.4Hz, 1H), 2.72 (ddd, J = 22.7, 11.9, 5.9 Hz, 2H), 2.62 – 2.58 (m, 1H), 2.44 –2.37 (m, 1H), 2.01 (ddd, J = 10.2, 5.1, 3.1 Hz, 1H).

[0108] Example 17: Synthesis of (S)-3-(5-(7-((2'-chloro-3-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-098)

[0109] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-3-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.74 (d, J =7.9 Hz, 1H), 7.68 (s, 1H), 7.60 – 7.55 (m, 2H), 7.48 – 7.42 (m, 4H), 7.31 (t, J= 8.9 Hz, 1H), 7.13 (s, 1H), 7.09 – 7.05 (m, 1H), 5.12 (dd, J = 13.3, 5.1Hz, 1H), 4.46 (d, J = 17.3 Hz, 1H), 4.34 (d, J = 17.3 Hz, 1H), 3.89 (t, J =5.4 Hz, 2H), 3.72 (d, J = 13.0 Hz, 1H), 3.40 (dd, J = 23.0, 11.0 Hz, 3H),2.92 (ddd, J = 17.5, 13.7, 5.4 Hz, 1H), 2.61 (dd, J = 12.4, 7.3 Hz, 3H), 2.44– 2.36 (m, 1H), 2.01 (dtd, J = 7.2, 5.1, 2.0 Hz, 1H).

[0110] Example 18: Synthesis of (S)-3-(5-(7-((2'-chloro-4-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-099)

[0111] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-4-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.76 (d, J =7.9 Hz, 1H), 7.74 (s, 1H), 7.64 (d, J = 8.0 Hz, 1H), 7.60 – 7.55 (m, 1H), 7.50 – 7.47 (m, 1H), 7.46 – 7.41 (m, 2H), 7.35 (dd,J = 7.0, 2.1 Hz, 1H),7.25 – 7.19 (m, 3H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 (d, J = 17.4 Hz, 1H), 4.36 (d, J = 17.4 Hz, 1H), 4.05 (t, J = 5.5 Hz, 2H), 3.60 (s, 2H), 3.49(dd, J = 38.1, 14.3 Hz, 2H), 2.95 – 2.88 (m, 1H), 2.78 – 2.67 (m, 2H), 2.63 –2.56 (m, 1H), 2.45 – 2.38 (m, 1H), 2.04 – 1.98 (m, 1H).

[0112] Example 19: Synthesis of (S)-3-(5-(7-((2'-chloro-5-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-100)

[0113] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-5-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.77 (d, J =7.9 Hz, 1H), 7.73 (s, 1H), 7.68 (dd, J = 8.6, 6.1 Hz, 1H), 7.63 (d, J = 8.0Hz, 1H), 7.60 – 7.56 (m, 1H), 7.47 – 7.41 (m, 2H), 7.39 (d, J = 6.7 Hz, 1H),7.31 – 7.24 (m, 2H), 7.06 (dd,J = 9.4, 2.7 Hz, 1H), 5.13 (dd, J = 13.3, 5.1Hz, 1H), 4.48 (d, J = 17.4 Hz, 1H), 4.36 (d, J = 17.4 Hz, 1H), 4.01 (t, J =5.2 Hz, 2H), 3.59 – 3.48 (m, 3H), 3.40 (d, J = 13.8 Hz, 1H), 2.92 (ddd, J =17.5, 13.7, 5.4 Hz, 1H), 2.69 (qd, J = 11.7, 6.2 Hz, 2H), 2.63 – 2.58 (m,1H), 2.45 – 2.38 (m, 1H), 2.01 (ddd, J = 10.2, 5.2, 3.1 Hz, 1H).

[0114] Example 20: Synthesis of (S)-3-(5-(7-((2'-chloro-6-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-101)

[0115] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-6-fluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 28 ClFN5O3 + [M+H] + Calculated value: 585.06; Measured value: 585.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.78 (d, J =7.9 Hz, 1H), 7.75 (s, 1H), 7.64 (d, J = 8.0 Hz, 1H), 7.62 – 7.59 (m, 1H), 7.51 (dd, J = 5.4, 3.3 Hz, 2H), 7.46 (ddd, J= 8.7, 7.4, 1.6 Hz, 2H), 7.42(d, J = 7.0 Hz, 1H), 7.30 – 7.23 (m, 2H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.49 (d, J = 17.4 Hz, 1H), 4.37 (d, J = 17.4 Hz, 1H), 4.03 (t, J = 5.0 Hz,2H), 3.59 (s, 2H), 3.47 (s, 2H), 2.95 – 2.88 (m, 1H), 2.77 – 2.67 (m, 2H),2.63 – 2.59 (m, 1H), 2.45 – 2.39 (m, 1H), 2.04 – 1.98 (m, 1H).

[0116] Example 21: Synthesis of (S)-3-(5-(7-((2'-chloro-3,4-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-102)

[0117] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-3,4-difluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 27 ClF2N5O3 + [M+H] + Calculated value: 603.05; Measured value: 603.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.70 (s, 1H), 7.62 – 7.55 (m, 2H), 7.53 – 7.49 (m, 1H), 7.48– 7.41 (m, 3H), 7.21 (s, 1H), 7.10 (dd, J = 8.3, 4.8 Hz, 1H), 5.13 (dd, J =13.3, 5.1 Hz, 1H), 4.47 (d, J= 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 3.91(t, J = 4.8 Hz, 2H), 3.79 (d, J = 13.0 Hz, 1H), 3.45 (dd, J = 51.1, 13.6 Hz,3H), 2.95 – 2.87 (m, 1H), 2.67 – 2.57 (m, 3H), 2.42 (dt, J = 13.6, 8.9 Hz,1H), 2.03 – 1.97 (m, 1H).

[0118] Example 22: Synthesis of (S)-3-(5-(7-((2'-chloro-3,6-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-103)

[0119] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-3,6-difluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 27 ClF2N5O3 + [M+H] + Calculated value: 603.05; Measured value: 603.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.77 (d, J = 7.9 Hz, 1H), 7.71 (s, 1H), 7.60 (dd, J = 10.6, 4.0 Hz, 2H), 7.53 – 7.44(m, 3H), 7.39 (ddq, J = 13.6, 9.0, 4.5 Hz, 2H), 7.28 (s, 1H), 5.13 (dd, J =13.3, 5.1 Hz, 1H), 4.48 (d, J = 17.4 Hz, 1H), 4.36 (d, J= 17.4 Hz, 1H), 3.96– 3.87 (m, 2H), 3.65 (d, J = 12.9 Hz, 1H), 3.51 – 3.38 (m, 3H), 2.92 (ddd, J = 17.4, 13.7, 5.4 Hz, 1H), 2.62 (dd, J = 17.6, 15.3 Hz, 3H), 2.45 – 2.38 (m,1H), 2.01 (dtd, J = 7.2, 5.1, 2.0 Hz, 1H).

[0120] Example 23: Synthesis of (S)-3-(5-(7-((2'-chloro-4,5-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-104)

[0121] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-4,5-difluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 27 ClF2N5O3 + [M+H] + Calculated value: 603.05; Measured value: 603.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.78 (d, J = 7.9 Hz, 1H), 7.75 (s, 1H), 7.68 (dd, J = 11.8, 8.5 Hz, 1H), 7.64 (d, J =8.0 Hz, 1H), 7.59 (dd, J = 7.8, 1.4 Hz, 1H), 7.45 (tdd, J = 8.8, 7.4, 1.6 Hz,2H), 7.39 (d, J = 6.7 Hz, 1H), 7.35 (dd, J = 11.0, 8.1 Hz, 1H), 7.30 (s, 1H), 5.13 (dd, J= 13.3, 5.1 Hz, 1H), 4.49 (d, J = 17.4 Hz, 1H), 4.37 (d, J = 17.4Hz, 1H), 4.08 – 4.01 (m, 2H), 3.63 (s, 2H), 3.51 – 3.41 (m, 2H), 2.95 – 2.88(m, 1H), 2.71 (qd, J = 11.9, 6.4 Hz, 2H), 2.63 – 2.59 (m, 1H), 2.45 – 2.38(m, 1H), 2.02 (ddd, J = 10.3, 5.2, 3.1 Hz, 1H).

[0122] Example 24: Synthesis of (S)-3-(5-(7-((2'-chloro-4,6-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-105)

[0123] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-4,6-difluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 27 ClF2N5O3 + [M+H] + Calculated value: 603.05; Measured value: 603.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.78 (d, J = 7.9 Hz, 1H), 7.75 (s, 1H), 7.65 (d, J = 7.9 Hz, 1H), 7.62 (dd, J = 7.9, 1.3 Hz, 1H), 7.48 (dtd, J = 16.7, 7.4, 1.6 Hz, 2H), 7.44 – 7.37 (m, 2H), 7.32(td, J = 9.2, 2.5 Hz, 1H), 7.27 (s, 1H), 5.13 (dd, J= 13.3, 5.1 Hz, 1H), 4.49 (d, J = 17.4 Hz, 1H), 4.37 (d, J = 17.4 Hz, 1H), 4.11 – 4.02 (m, 2H), 3.63 (s, 2H), 3.45 (t, J = 17.8 Hz, 2H), 2.92 (ddd, J = 17.4, 13.7, 5.4 Hz,1H), 2.73 (qd, J = 11.7, 6.3 Hz, 2H), 2.63 – 2.59 (m, 1H), 2.45 – 2.38 (m,1H), 2.02 (dtd, J = 7.2, 5.0, 2.0 Hz, 1H).

[0124] Example 25: Synthesis of (S)-3-(5-(7-((2'-chloro-5,6-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-106)

[0125] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-5,6-difluoro-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 27 ClF2N5O3 + [M+H] + Calculated value: 603.05; Measured value: 603.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.78 (d, J = 7.9 Hz, 1H), 7.74 (s, 1H), 7.63 (d, J = 7.4 Hz, 2H), 7.57 – 7.47 (m, 5H), 7.27 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.49 (d, J = 17.4 Hz, 1H), 4.37 (d, J = 17.4 Hz, 1H), 4.01 (t,J = 5.1 Hz, 2H), 3.58 (s, 2H), 3.43 (d, J = 17.0 Hz, 2H), 2.95 – 2.87 (m, 1H), 2.73 – 2.65 (m, 2H), 2.61 (dd, J = 10.3,6.4 Hz, 1H), 2.45 – 2.38 (m, 1H), 2.02 (ddd, J = 10.2, 5.2, 3.1 Hz, 1H).

[0126] Example 26: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-139)

[0127]

[0128] Step 1: Referring to Step 3 of Example 1, 3-bromo-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 3-bromo-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 27 H 37 N6O6 + [M+H] + Calculated value: 541.63; Measured value: 541.60.

[0129] Step 2: Synthesize the target product according to the method in step 4 of Example 1. LCMS (ESI) C 18 H 19 N6O3 + [M+H] + Calculated value: 367.39; Measured value: 367.40.

[0130] Step 3: Referring to the method in step 5 of Example 1, ( S The reaction of 3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione with 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde yielded the target product (32.00 mg, 43.99% yield). LCMS (ESI) C 31 H 28ClN6O3 + [M+H] + Calculated value: 568.06; Measured value: 568.10. 1 H NMR (600 MHz, DMSO) δ 11.01 (s, 1H), 8.01 (s, 1H), 7.90 (d, J = 7.9 Hz, 1H), 7.86 (d, J = 7.9 Hz, 1H), 7.65 (d, J = 7.7 Hz, 1H),7.59 – 7.53 (m, 1H), 7.45 (td, J = 6.3, 3.2 Hz, 1H), 7.43 – 7.41 (m, 2H), 7.40 – 7.38 (m, 1H), 7.37 – 7.35 (m, 1H), 7.18 (dd, J = 7.5, 1.0 Hz, 1H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.54 (d, J = 17.5 Hz, 1H), 4.42 (d, J = 17.5Hz, 1H), 4.10 (t, J = 5.4 Hz, 2H), 3.65 (s, 2H), 3.58 (d, J = 13.8 Hz, 1H), 3.48 (d, J = 13.8 Hz, 1H), 2.96 – 2.89 (m, 1H), 2.72 (tdd, J = 18.2, 11.8,6.2 Hz, 2H), 2.64 – 2.60 (m, 1H), 2.45 – 2.39 (m, 1H), 2.04 (ddd, J = 10.2, 5.1, 3.1 Hz, 1H).

[0131] Example 27: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-8-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-146)

[0132]

[0133] Step 1: Dissolve 3-methylpyrazine-2-amine (1.0 eq) and 2-chloroacetaldehyde (1.5 eq) in ethanol, then add 37% concentrated hydrochloric acid to adjust the pH of the reaction system to 3. Heat the reaction mixture to 75°C and stir for 12 h. Detect the completion of the reaction by LC-MC or TLC, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-126 (4.70 g, yield 77.04%). LC-MS (ESI) C7H8N3 + [M+H] + Calculated value: 134.16; Measured value: 134.20.

[0134] Step 2: Synthesis of 8-methyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine (LC-05-135)

[0135] LC-05-126 (1.0 eq) was dissolved in ethylene glycol monomethyl ether. The solution was purged three times with hydrogen, and 10% Pd / C (0.20 eq) was added, followed by another three purgings with hydrogen. The reaction mixture was stirred at room temperature for 30 h. The reaction was monitored for completion by LC-MC or TLC. The reaction mixture was filtered through diatomaceous earth to remove Pd / C, washed several times with ethanol, and the filtrate was collected and concentrated under reduced pressure to obtain a white, viscous LC-05-135 (4.8 g, 99.13% yield). This crude product was directly added to the next step without purification. LC-MS (ESI) C7H 12 N3 + [M+H] + Calculated value: 138.20; Measured value: 138.20.

[0136] Step 3: Dissolve LC-05-135 (1.0 eq) in dichloromethane, then add N,N-diisopropylethylamine (2 eq) and di-tert-butyl dicarbonate (1.5 eq). Stir the reaction mixture at room temperature for 12 h. Detect the completion of the reaction by LCMC or TLC. Add water to the reaction mixture, extract with dichloromethane, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white, viscous LC-05-138 (5.56 g, yield 66.96%). LC-MS (ESI) C 12 H 20 N3O2 + [M+H] + Calculated value: 238.31; Measured value: 238.30.

[0137] Step 4: Dissolve LC-05-138 (1.0 eq) in acetonitrile, then add N-bromosuccinimide (1.0 eq). Stir the reaction mixture at room temperature for 12 h. Detect the completion of the reaction by LCMC or TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-142 (2.30 g, yield 61.65%). LC-MS (ESI) C 12 H 19 BrN3O2 + [M+H] + Calculated values: 316.07 and 318.07; measured values: 316.10 and 318.10.

[0138] Step 5: Referring to the method in Step 3 of Example 1, 3-bromo-5,6-dihydroimidazole[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 3-bromo-8-methyl-5,6-dihydroimidazole[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 29 H 40 N5O6 + [M+H] + Calculated value: 554.67; Measured value: 554.70.

[0139] Step 6: Refer to the method in step 4 of Example 1. LCMS (ESI) C 20 H 22 N5O3 + [M+H] + Calculated value: 380.17; Measured value: 380.20.

[0140] Step 7: Referring to the method in Step 5 of Example 1, replace 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 33 H 31 ClN5O3 + [M+H] + Calculated value: 580.21; Measured value: 580.20.

[0141] Example 28: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-164)

[0142]

[0143] Step 1: Following the method in Step 3 of Example 27, replace 8-methyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine with 5,6,7,8-tetrahydroimidazole[1,5-a]pyrazine to synthesize the target product. LCMS (ESI) C 11 H 18 N3O2 + [M+H] + Calculated value: 224.29; Measured value: 224.30.

[0144] Step 2: Dissolve LC-05-151 (1.0 eq) in tetrahydrofuran, lower the reaction temperature to -78°C, and then add n-butyllithium (1.2 eq, 2.5 M in Heptane). Stir the reaction mixture at -78°C for 1 h. Then, add iodine (1.2 eq) in tetrahydrofuran dropwise to the reaction mixture, and continue stirring at -78°C for another 1 h. Detect the completion of the reaction using LC-MC or TLC. Quench the reaction mixture with saturated ammonium chloride solution, extract with ethyl acetate, wash the organic phase with saturated sodium thiosulfate solution, saturated sodium bicarbonate solution, water, and saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain an orange solid LC-05-158 (1.13 g, yield 32.84%). LC-MS (ESI) C 11 H 17 IN3O2 + [M+H] + Calculated value: 350.18; Measured value: 350.20.

[0145] Step 3: Referring to the method in Step 3 of Example 1, 3-bromo-5,6-dihydroimidazole[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 3-iodo-5,6-dihydroimidazole[1,5-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 28 H 38 N5O6 + [M+H] + Calculated value: 540.64; Measured value: 540.60.

[0146] Step 4: Refer to the method in step 4 of Example 1. LCMS (ESI) C 19 H 20 N5O3 + [M+H] +Calculated value: 366.40; Measured value: 366.40.

[0147] Step 5: Referring to the method in Step 5 of Example 1, replace 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 32 H 29 ClN5O3 + [M+H] + Calculated value: 567.07; Measured value: 567.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.92 (s, 1H), 7.85 (d, J = 8.2 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.65 (d, J = 7.6 Hz, 1H),7.57 – 7.54 (m, 1H), 7.46 – 7.40 (m, 3H), 7.39 – 7.35 (m, 2H), 7.17 (dd, J =7.5, 0.7 Hz, 1H), 6.79 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.50 (d, J =17.4 Hz, 1H), 4.38 (d, J = 17.3 Hz, 1H), 4.13 (t, J = 5.4 Hz, 2H), 3.54 – 3.39 (m, 4H), 2.96 – 2.89 (m, 1H), 2.71 – 2.65 (m, 1H), 2.64 – 2.57 (m, 2H), 2.45 – 2.38 (m, 1H), 2.05 – 2.00 (m, 1H).

[0148] Example 29: Synthesis of (S)-3-(5-(7-(1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-176)

[0149] (S)-3-(1-oxo-5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidin-2,6-dione (1.0 eq) and 2-(1-bromoethyl)-2'-chloro-1,1'-biphenyl (1.0 eq) were dissolved in N,N-dimethylformamide solution, and then K2CO3 (2.0 eq) was added. The reaction solution was heated to 80 °C and stirred for 12 h. The reaction was monitored by LCMS until completion, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-TLC on silica gel plates to give a white solid (S)-3-(5-(7-(1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethyl)-5,6,7,8-tetrahydroimidazol[1,2-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (6.00 mg, yield 23.25%). LC-MS (ESI) C 33 H 31 ClN5O3 + [M+H] + Calculated value: 581.10; Measured value: 581.15.

[0150] Example 30: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-3-methyl-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-186)

[0151]

[0152] Step 1: Dissolve 4-bromo-2-ethylbenzoic acid (1.0 eq) in N,N-dimethylformamide solution, then add iodomethane (2.0 eq) and Cs₂CO₃ (2.0 eq). Heat the reaction mixture to 85 °C and stir for 15 h. Detect the reaction completion by TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-167 (5.00 g, yield 94.23%). LC-MS (ESI) C 10 H 12 BrO2 + [M+H] + Calculated values: 243.11 and 245.23; measured values: 243.10 and 245.30.

[0153] Step 2: Dissolve LC-05-167 (1.0 eq) in benzene solution, then add NBS (1.5 eq) and AIBN (0.30 eq). Stir the reaction mixture at room temperature for 12 h. Detect the completion of the reaction by TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-168 (5.00 g, yield 75.75%). LC-MS (ESI) C 10 H 11 Br2O2 + [M+H] + Calculated values: 320.91 and 322.91; measured values: 321.10 and 323.30.

[0154] Step 3: Dissolve LC-05-168 (1.0 eq) and (S)-4,5-diamino-5-oxovalerate tert-butyl hydrochloride (1.0 eq) in acetonitrile, then add N,N-diisopropylethylamine (3 eq). Heat the reaction mixture to 85 °C and stir for 12 h. Detect the completion of the reaction by LCMS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-175 (4.20 g, yield 48.75%). LC-MS (ESI) C 18 H 24 BrN2O4 + [M+H] + Calculated values: 411.09 and 413.09; measured values: 411.10 and 413.10.

[0155] Step 4: Dissolve LC-05-175 (1 eq) and pinacol diborate (1.5 eq) in 1,4-dioxane, and add potassium acetate (3 eq). Purge the solution three times with nitrogen, add Pd(dppf)Cl2 (0.03 eq), and purge again three times with nitrogen. Heat the reaction mixture to 100°C and stir for 12 h. Detect the reaction completion by LCMS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-179 (1.50 g, yield 74.78%). LC-MS (ESI)C 24 H 36 BN2O6 + [M+H] + Calculated value: 459.27; Measured value: 459.30.

[0156] Step 5: Dissolve LC-05-179 (1 eq) and 3-bromo-5,6-dihydroimidozoprothiolane[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) in 1,4-dioxane, and add 2 M sodium carbonate solution (2 eq). Purge three times with nitrogen, add Pd(PPh3)4 (0.04 eq), and purge three more times with nitrogen. Heat the reaction mixture to 85 °C and stir for 12 h. Detect the completion of the reaction using LC-MS, and cool the reaction mixture to room temperature. Add water and ethyl acetate to the reaction mixture for extraction, repeating the extraction three times until complete. Wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-181 (580.00 mg, yield 87.30%). LC-MS (ESI)C 29 H 40 N5O6 + [M+H] + Calculated value: 554.67; Measured value: 554.70.

[0157] Step 6: Dissolve LC-05-181 (1 eq) in acetonitrile and add benzenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction mixture to room temperature. Concentrate under reduced pressure to remove the solvent. Add ethyl acetate and petroleum ether to the residue and slurry. Filter to obtain a white solid LC-05-184 (350.00 mg, yield 88.06%). LC-MS (ESI) C 20 H 22 N5O3 + [M+H] + Calculated value: 380.43; Measured value: 380.40.

[0158] Step 7: Dissolve LC-05-184 and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) in... N , N A catalytic amount of acetic acid was added to dimethylformimide. The reaction mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium borohydride acetate (3 eq), and stirring was continued for 12 h. The reaction was monitored for completion by LC-MS, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate analysis to give LC-05-186 (23.00 mg, yield 30.09%) as a white solid. LC-MS (ESI) C 33 H 31ClN5O3 + [M+H] + Calculated value: 581.10; Measured value: 581.15.

[0159] Example 31: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-3-methyl-1-oxoisoindololin-2-yl)piperidine-2,6-dione (LC-05-187)

[0160]

[0161] Step 1: Referring to the method in Step 5 of Example 30, 3-bromo-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 3-bromo-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 28 H 39 N6O6 + [M+H] + Calculated value: 555.66; Measured value: 555.70.

[0162] Step 2: Refer to the method in step 6 of Example 30. LCMS (ESI) C 19 H 21 N6O3 + [M+H] + Calculated value: 381.17; Measured value: 381.20.

[0163] Step 3: Referring to the method in Step 7 of Example 30, (S)-3-(3-methyl-1-oxo-5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione was replaced with (S)-3-(3-methyl-1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione to synthesize the target product. LCMS (ESI) C 32 H 30 ClN6O3 + [M+H] + Calculated value: 581.21; Measured value: 581.20.

[0164] Example 32: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-190)

[0165]

[0166] Step 1: Dissolve 6-methylpyrazine-2-amine (1.0 eq) and 2-chloroacetaldehyde (1.5 eq) in ethanol, then add 37% concentrated hydrochloric acid to adjust the pH of the reaction system to 3. Heat the reaction mixture to 75°C and stir for 48 h. Detect the completion of the reaction by LC-MC or TLC, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-148 (3.10 g, yield 50.81%). LC-MS (ESI) C7H8N3 + [M+H] + Calculated value: 134.16; Measured value: 134.20.

[0167] Step 2: Dissolve LC-05-148 (1.0 eq) in ethylene glycol monomethyl ether. Purge three times with hydrogen, add 10% Pd / C (0.20 eq), and purge three more times with hydrogen. Heat the reaction mixture to 60°C and stir for 30 h. Detect the reaction completion by LCMC or TLC. Filter the reaction mixture through diatomaceous earth to remove Pd / C, wash several times with ethanol, collect the filtrate, and concentrate under reduced pressure to obtain a white, viscous LC-05-174 (3.00 g, yield 97.06%). This crude product was directly added to the next step without purification. LC-MS (ESI)C7H 12 N3 + [M+H] + Calculated value: 138.20; Measured value: 138.20.

[0168] Step 3: Dissolve LC-05-174 (1.0 eq) in dichloromethane, then add N,N-diisopropylethylamine (2 eq) and di-tert-butyl dicarbonate (1.5 eq). Stir the reaction mixture at room temperature for 12 h. Detect the completion of the reaction by LCMC or TLC. Add water to the reaction mixture, extract with dichloromethane, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white, viscous LC-05-180 (2.90 g, yield 46.57%). LC-MS (ESI) C 12 H20 N3O2 + [M+H] + Calculated value: 238.31; Measured value: 238.30.

[0169] Step 4: Dissolve LC-05-180 (1.0 eq) in acetonitrile, then add N-bromosuccinimide (1.0 eq). Stir the reaction mixture at room temperature for 12 h. Detect the completion of the reaction by LCMC or TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-183 (1.75 g, yield 82.08%). LC-MS (ESI) C 12 H 19 BrN3O2 + [M+H] + Calculated value: 317.21; Measured value: 317.20.

[0170] Step 5: Referring to the method in Step 3 of Example 1, 3-bromo-5,6-dihydroimidazole[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 3-bromo-5-methyl-5,6-dihydroimidazole[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 29 H 40 N5O6 + [M+H] + Calculated value: 554.30; Measured value: 554.30.

[0171] Step 6: Synthesize the target product according to the method in step 4 of Example 1. LCMS (ESI) C 20 H 22 N5O3 + [M+H] + Calculated value: 380.17; Measured value: 380.20.

[0172] Step 7: Referring to the method in Step 5 of Example 1, replace 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 33 H 31 ClN5O3 + [M+H] + Calculated value: 580.21; Measured value: 580.20. 1H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.76 (d, J =7.9 Hz, 1H), 7.72 (d, J = 4.3 Hz, 1H), 7.64 – 7.60 (m, 2H), 7.56 (ddd, J =8.9, 6.7, 2.7 Hz, 1H), 7.46 – 7.40 (m, 3H), 7.37 (t, J = 7.4 Hz, 1H), 7.35 –7.32 (m, 1H), 7.17 (d, J = 7.4 Hz, 1H), 7.04 (s, 1H), 5.13 (dd, J = 13.2, 4.8Hz, 1H), 4.63 (ddd, J = 13.9, 6.8, 3.6 Hz, 1H), 4.48 (dd, J = 17.3, 8.7 Hz, 1H), 4.36 (dd, J = 17.3, 5.5 Hz, 1H), 3.68 (dd, J = 45.6, 14.6 Hz, 1H), 3.55– 3.49 (m, 1H), 3.43 (dd, J = 13.4, 9.2 Hz, 1H), 3.22 (dd, J = 27.6, 14.6 Hz,1H), 2.96 – 2.89 (m, 1H), 2.77 – 2.64 (m, 2H), 2.62 – 2.59 (m, 1H), 2.44 –2.36 (m, 1H), 2.07 – 1.97 (m, 1H), 0.90 (d, J = 6.3 Hz, 3H).

[0173] Example 33: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-8,8-dimethyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-05-200)

[0174]

[0175] Step 1: Dissolve 1.0 eq of 2-(benzyloxy)carbonyl)amino)-2-methylpropionic acid in dichloromethane, purge three times with nitrogen, and stir the reaction mixture at 0°C for 30 minutes. Then add 1.20 eq of N,O-dimethylhydroxylamine hydrochloride, 1.50 eq of HATU, and 4.0 eq of triethylamine, purge three more times with nitrogen, and stir the reaction mixture at 0°C for another 30 minutes. Heat the reaction mixture to room temperature and stir for 16 h. Detect the completion of the reaction by LC-MS. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain 11.75 g (82.87%) as a white solid. LC-MS (ESI) C 14 H 21 N2O4 + [M+H] + Calculated value: 281.15; Measured value: 281.20.

[0176] Step 2: Dissolve LC-05-122 (1.0 eq) in tetrahydrofuran, purge with nitrogen three times, and stir the reaction mixture at -78°C for 10 minutes. Then, add LiAlH4 (1.0 eq, 1M tetrahydrofuran solution) dropwise, and continue stirring at -78°C for 40 minutes. Detect the completion of the reaction using LC-MC. Quench the reaction mixture by adding ethyl acetate dropwise at -78°C. Saturated sodium sulfate is then slowly added dropwise, and the mixture is stirred at room temperature for 15 minutes. Filter the reaction mixture with diatomaceous earth, wash with ethyl acetate, and concentrate the filtrate under reduced pressure to obtain a white solid, LC-05-124 (3.0 g, yield 69.11%). This compound was added directly to the next step without purification. LC-MS (ESI) C 12 H 16 NO3 + [M+H] + Calculated value: 222.12; Measured value: 222.10.

[0177] Step 3: Dissolve LC-05-124 (1.0 eq) in methanol. Stir the reaction mixture at 0°C for 10 minutes, then add 25% ammonia solution and 40% glyoxal solution. Stir the reaction mixture at room temperature for 48 h. Detect the completion of the reaction using LC-MS. Concentrate the reaction mixture under vacuum to remove the organic solution. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-125 (7.70 g, yield 77.29%). LC-MS (ESI) C 14 H 17 N3O2+ [M+H] + Calculated value: 260.14; Measured value: 260.20.

[0178] Step 4: Dissolve LC-05-125 (1.0 eq) in DMF, then add ethyl 2-bromoacetate (1.50 eq) and K₂CO₃ (2.0 eq). Continue stirring the reaction mixture at 25°C for 15 h. Detect the completion of the reaction using LC-MC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-05-131 (8.00 g, yield 78.00%). LC-MS (ESI) C 18 H 24 N3O4 + [M+H] + Calculated value: 346.17; Measured value: 346.20.

[0179] Step 5: Dissolve LC-05-131 (1.0 eq) in methanol, then add 10% Pd / C. Stir the reaction mixture at room temperature under hydrogen atmosphere for 15 h. Detect the reaction completion by LC-MC. Filter the reaction mixture through diatomaceous earth and wash repeatedly with methanol. Concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography to obtain a white, viscous LC-05-154 (2.50 g, yield 65.34%). LC-MS (ESI) C8H 12 N3O + [M+H] + Calculated value: 166.10; Measured value: 166.10.

[0180] Step 6: Dissolve LC-05-154 (1.0 eq) in tetrahydrofuran, then add diphenylsilane (1.0 eq), purge with nitrogen for 10 minutes, then add RhCO(PPh3)3 (0.50 eq), and stir the reaction mixture at 25°C for 16 h. The reaction is complete as determined by LC-MC. Add 1M hydrochloric acid solution to the reaction mixture, extract with ethyl acetate, then adjust the pH of the aqueous phase with saturated sodium bicarbonate, extract with ethyl acetate multiple times, wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography to obtain a white, viscous LC-05-160 (1.15 g, yield 62.82%). LC-MS (ESI) C8H 14 N3 + [M+H] + Calculated value: 151.21; Measured value: 151.20.

[0181] Step 7: Referring to the method in Step 3 of Example 33, replace 5-methyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine with 8,8-dimethyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine to synthesize the target product. LCMS (ESI)C 13 H 22 N3O2 + [M+H] + Calculated value: 252.33; Measured value: 252.30.

[0182] Step 8: Referring to the method in Step 4 of Example 32, 5-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 8,8-dimethyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 13 H 21 BrN3O2 + [M+H] + Calculated values: 330.07 and 332.07; measured values: 330.10 and 332.10.

[0183] Step 9: Referring to Step 3 of Example 1, replace 3-bromo-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester with 3-bromo-8,8-dimethyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 30 H 42 N5O6 + [M+H] + Calculated value: 568.31; Measured value: 568.30.

[0184] Step 10: Referring to the method in Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-8,8-dimethyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS(ESI) C 21 H 24 N5O3 + [M+H] + Calculated value: 394.45; Measured value: 394.50.

[0185] Step 11: Dissolve (S)-3-(5-(8,8-dimethyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (1.0 eq) in DMF, then add NaI (0.50 eq) and K2CO3 (2.0 eq), and continue stirring the reaction solution at 80 °C for 12 h. The reaction was monitored by LC-MS to confirm completion. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give LC-05-200 (21.00 mg, yield 39.81%) as a white solid. LC-MS (ESI)C 34 H 33 ClN5O3 + [M+H] + Calculated value: 595.11; Measured value: 595.20.

[0186] Example 34: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-002)

[0187]

[0188] Step 1: Dissolve 1.0 eq of 4-oxopiperidinium-1-carboxylic acid tert-butyl ester in DMF, then add 2.50 eq of DMA-DMF. Stir the reaction mixture at 153 °C for 4 h. LC-MS was used to detect the completion of the reaction. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a brownish-yellow oil, LC-05-178 (845.00 mg, yield 66.20%). LC-MS (ESI) C 13 H 23 N2O3 + [M+H] + Calculated value: 255.33; Measured value: 255.30.

[0189] Step 2: Dissolve ZPP-02-107 (1.0 eq) in 6M hydrochloric acid solution, then add NaNO2 solution, and continue stirring the reaction solution at -5℃ for 1 h. Dissolve SnCl2 in concentrated hydrochloric acid, add the prepared SnCl2 solution to the reaction solution, and continue stirring the reaction solution at 25℃ for 2 h. Detect the completion of the reaction using LC-MS. Concentrate the reaction solution under reduced pressure, dissolve the residue in ethanol, filter and concentrate under reduced pressure to obtain a yellow solid LC-05-177 (3.12 g, yield 58.98%). LC-MS (ESI) C 13 H 15 N4O3 + [M+H] + Calculated value: 275.28; Measured value: 275.30.

[0190] Step 3: Acetic acid (5.00 eq), the reaction mixture was stirred at 25 °C for 1 h. The reaction was monitored by LC-MC to confirm completion. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give LC-05-197 (432.00 mg, yield 31.82%) as a white solid. LC-MS (ESI) C 24 H 28 N5O5 + [M+H] + Calculated value: 466.51; Measured value: 466.50.

[0191] Step 4: Dissolve LC-05-197 (1.0 eq) in dichloromethane, then add 4M ethyl acetate hydrochloride solution. Continue stirring the reaction mixture at 25°C for 1 h. Detect the completion of the reaction using LC-MC. Concentrate the reaction mixture under reduced pressure to obtain a white solid, LC-06-001 (250.00 mg, yield 99.53%). LC-MS (ESI) C 19 H 20 N5O3 + [M+H] + Calculated value: 366.39; Measured value: 366.40.

[0192] Step 5: Referring to the method in Step 3 of Example 26, replace (S)-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione with (S)-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)isoindololin-2-yl)piperidine-2,6-dione to synthesize the target product. LCMS (ESI) C32 H 29 ClN5O3 + [M+H] + Calculated value: 567.06; Measured value: 567.10.

[0193] Example 35: Synthesis of 5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (LC-06-004)

[0194]

[0195] Step 1: Synthesis of 5-bromo-2-(2,6-dioxopiperidin-3-yl)isodihydroindole-1,3-dione (ZPP-02-047)

[0196] 4-Bromophthalic anhydride (2 g, 1 eq) and 3-amino-2,6-piperidinidone hydrochloride (1.4 eq) were dissolved in acetic acid, followed by the addition of sodium acetate (1.7 eq). The reaction mixture was heated to 120 °C and stirred for 4 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give ZPP-02-047 (2.9 g, 97.64% yield). LC-MS (ESI) C 13 H9BrN2O4 + [M+H] + Calculated value: 337.13; Measured value: 335.97 and 337.97.

[0197] Step 2: ZPP-02-047 (2.9 g, 1 eq) and pinacol diborate (1.5 eq) were dissolved in 1,4-dioxane, and potassium acetate (3 eq) was added. The mixture was purged with nitrogen three times, and Pd(dppf)Cl2 (0.03 eq) was added. The mixture was then purged with nitrogen three times again. The reaction mixture was heated to 110 °C and stirred for 15 h. The reaction was monitored by LC-MS to confirm completion, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-02-052 (2.3 g, yield 69.70%). LC-MS (ESI) C 19 H 21 BN2O6 + [M+H] +Calculated value: 384.19; Measured value: 385.15.

[0198] Step 3: ZPP-02-052 (2.3 g, 1 eq) and 3-bromo-5,6-dihydroimidazolo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) were dissolved in a tetrahydrofuran:water = 20:1 mixture, and tripotassium phosphate (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(dtbpf)Cl2 (0.05 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 80 °C and stirred for 3 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-02-096 (1.00 g, yield 34.84%). LC-MS (ESI) C 24 H 25 N5O6 + [M+H] + Calculated value: 479.49; Measured value: 480.18.

[0199] Step 4: Referring to the method in Step 4 of Example 34, (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester was replaced with 3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 19 H 18 N5O4 + [M+H] + Calculated value: 380.38; Measured value: 380.40.

[0200] Step 5: Referring to the method in Step 3 of Example 26, replace (S)-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindolin-2-yl)piperidin-2,6-dione with 2-(2,6-dioxopiperidin-3-yl)-5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindolin-1,3-dione to synthesize the target product. LCMS(ESI) C 32 H 27 ClN5O4 + [M+H] +Calculated value: 581.04; Measured value: 581.10. 1 H NMR (600 MHz, DMSO) δ11.14 (s, 1H), 7.99 (s, 1H), 7.98 – 7.92 (m, 2H), 7.67 (d, J = 7.6 Hz, 1H),7.59 – 7.54 (m, 1H), 7.46 – 7.40 (m, 3H), 7.40 – 7.35 (m, 3H), 7.17 (dd, J =7.6, 1.0 Hz, 1H), 5.17 (dd, J = 12.9, 5.4 Hz, 1H), 4.12 – 4.02 (m, 2H), 3.57– 3.50 (m, 3H), 3.45 (d, J = 13.8 Hz, 1H), 2.90 (dt, J = 11.8, 4.3 Hz, 1H),2.75 – 2.67 (m, 2H), 2.63 – 2.59 (m, 1H), 2.56 – 2.51 (m, 1H), 2.07 (ddd, J =10.4, 5.4, 3.1 Hz, 1H).

[0201] Example 36: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-014)

[0202]

[0203] Step 1: ZPP-02-115 (1.0 eq) was dissolved in tetrahydrofuran, then acetic acid (1.10 eq) and tert-butyl nitrite (1.50 eq) were added. The reaction mixture was stirred at 100 °C for 12 h. The reaction was detected by LC-MS. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give LC-06-007 (796.00 mg, yield 90.23%) as a white solid. LC-MS (ESI) C 22 H 25 N6O4 + [M+H] +Calculated value: 437.47; Measured value: 437.50.

[0204] Step 2: Dissolve LC-06-007 (1.0 eq) in methanol. Purge three times with hydrogen, then add 10% Pd / C (0.20 eq), and purge three more times with hydrogen. Stir the reaction mixture at room temperature for 12 h. Detect the reaction completion by LC-MS. Filter the reaction mixture through diatomaceous earth to remove Pd / C, wash several times with methanol, collect the filtrate, and concentrate under reduced pressure to obtain a white solid LC-06-012 (543.00 mg, yield 61.14%). This crude product was directly added to the next step without purification. LC-MS (ESI) C 22 H 29 N6O4 + [M+H] + Calculated value: 441.50; Measured value: 441.50.

[0205] Step 3: Referring to the method in Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-5-amino-5-oxo-4-(1-oxo-5-(4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)isoindoline-2-yl)valerate tert-butyl ester to synthesize the target product. LCMS (ESI) C 18 H 19 N6O3 + [M+H] + Calculated value: 367.38; Measured value: 367.40.

[0206] Step 4: Referring to the method in step 3 of Example 26, ( S The target product (23.00 mg, yield 29.72%) was synthesized by replacing (S)-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione with (S)-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)isoindololin-2-yl)piperidine-2,6-dione. LCMS (ESI) C 31 H 28 ClN6O3 + [M+H] + Calculated value: 568.05; Measured value: 568.10. 1H NMR (600 MHz, DMSO) δ 11.02 (s, 1H), 7.94 (s, 1H), 7.92 (d, J =8.1 Hz, 1H), 7.81 (dd, J = 8.1, 1.6 Hz, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.56 –7.54 (m, 1H), 7.43 – 7.40 (m, 3H), 7.38 – 7.34 (m, 2H), 7.16 (dd, J = 7.5, 1.1 Hz, 1H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.56 (d, J = 17.7 Hz, 1H), 4.44(d, J = 17.7 Hz, 1H), 3.57 (d, J = 13.9 Hz, 1H), 3.47 (d, J = 13.5 Hz, 3H), 2.92 (ddd, J = 17.5, 13.7, 5.4 Hz, 1H), 2.84 (t, J = 4.8 Hz, 2H), 2.65 – 2.58(m, 3H), 2.47 – 2.40 (m, 1H), 2.05 – 2.02 (m, 1H).

[0207] Example 37: Synthesis of (S)-3-(5-(7-((3-(2-chlorophenyl)pyridin-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-017)

[0208] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 3-(2-chlorophenyl)pyridinaldehyde. LCMS (ESI) C 31 H 28 ClN6O3 + [M+H] + Calculated value: 568.06; Measured value: 568.10.

[0209] Example 38: Synthesis of (S)-3-(5-(7-((4-(2-chlorophenyl)pyridin-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-018)

[0210] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 4-(2-chlorophenyl)pyridinaldehyde. LCMS (ESI) C 31 H 28 ClN6O3 + [M+H] + Calculated value: 568.06; Measured value: 568.10.

[0211] Example 39: Synthesis of (S)-3-(5-(7-((3-(2-chlorophenyl)pyridin-4-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-019)

[0212] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 3-(2-chlorophenyl)isonicotinic acid aldehyde. LCMS (ESI) C 31 H 28 ClN6O3 + [M+H] + Calculated value: 568.06; Measured value: 568.10. 1 H NMR (600 MHz, DMSO) δ 11.02 (s, 1H), 8.69 (d, J = 5.1 Hz,1H), 8.44 (s, 1H), 7.92 (s, 1H), 7.91 – 7.85 (m, 2H), 7.75 (dd, J = 13.4, 6.7Hz, 2H), 7.64 (dd, J = 7.8, 1.0 Hz, 1H), 7.53 – 7.47 (m, 2H), 7.44 (dd, J =7.4, 1.7 Hz, 1H), 5.16 (dd, J = 13.3, 5.1 Hz, 1H), 4.53 (d, J = 17.6 Hz, 1H), 4.42 (d,J = 17.6 Hz, 1H), 4.17 (t, J = 4.9 Hz, 2H), 3.94 (s, 2H), 3.67 (s,2H), 2.96 – 2.90 (m, 1H), 2.86 (t, J = 5.2 Hz, 2H), 2.64 – 2.60 (m, 1H), 2.46 – 2.40 (m, 1H), 2.07 – 2.00 (m, 1H).

[0213] Example 40: Synthesis of (S)-3-(5-(7-((2-(2-chlorophenyl)pyridin-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-027)

[0214] Referring to step 5 of Example 1, [1,1'-biphenyl]-2-carboxaldehyde was replaced with 2-(2-chlorophenyl)nicotinaldehyde to synthesize the target product. LCMS (ESI) C 31 H 28 ClN6O3 + [M+H] + Calculated value: 568.06; Measured value: 568.10.

[0215] Example 41: Synthesis of (S)-3-(5-(7-((5-(2-chlorophenyl)pyrimidin-4-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-042)

[0216] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 5-(2-chlorophenyl)pyrimidin-4-carboxaldehyde. LCMS (ESI) C 30 H 27 ClN7O3 + [M+H] + Calculated value: 569.04; Measured value: 569.00.

[0217] Example 42: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-imidazol[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-045)

[0218]

[0219] Step 1: ZPP-02-106 (1 eq) was dissolved in acetic acid, and sodium acetate (2 eq) and 4-chloro-3-nitropyridine (2 eq) were added. The reaction mixture was reacted at 50 °C for 12 h. After the reaction was completed as detected by LC-MS, water was added to quench the reaction, the pH was adjusted to approximately 7 with saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-02-110 (2.94 g, yield 71.74%). LC-MS (ESI) C 22 H 25 N5O6 + [M+H] + Calculated value: 455.47; Measured value: 456.18.

[0220] Step 2: Dissolve ZPP-02-110 (1 eq) in methanol and add palladium on carbon (10% W). Replace with hydrogen three times, and stir the reaction mixture at room temperature for 3 h. After the reaction is complete, monitor the reaction by LC-MS. Filter with diatomaceous earth, and evaporate the filtrate to dryness to obtain ZPP-02-115 (2.4 g, yield 85.71%). LC-MS (ESI) C 22 H 27 N5O4 + [M+H] + Calculated value: 425.49; Measured value: 426.21.

[0221] Step 3: Dissolve ZPP-02-115 (1 eq) in trimethyl orthoformate and add p-toluenesulfonic acid (0.06 eq). Heat the reaction mixture to 100°C and stir for 1 h. After the reaction is complete as detected by LC-MS, quench with water, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-02-126 (1 g, yield 65.36%). LC-MS (ESI) C 23 H 25 N5O4 + [M+H] + Calculated value: 435.48; Measured value: 436.19.

[0222] Step 4: Dissolve ZPP-02-126 (1 eq) in dichloromethane and add benzyl bromide. Stir the reaction mixture at room temperature for 4 h. Concentrate the reaction mixture under reduced pressure, then dissolve it in methanol, and slowly add sodium borohydride. React for 5 h. After the reaction is complete as detected by LC-MS, quench with water, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-02-134 (300 mg, yield 52.07%). LC-MS (ESI) C 30 H 35 N5O4 + [M+H] + Calculated value: 529.64; Measured value: 530.27.

[0223] Step 5: Dissolve ZPP-02-134 (1 eq) in methanol and add palladium on carbon (10% W). Replace with hydrogen three times, and stir the reaction mixture at room temperature for 12 h. After the reaction is complete, detect the reaction by LC-MS. Filter with diatomaceous earth, and evaporate the filtrate to dryness to obtain ZPP-02-143 (260 mg, yield 89.65%). LC-MS (ESI) C 23 H 29 N5O4 + [M+H] + Calculated value: 439.52; Measured value: 440.23.

[0224] Step 6: Dissolve ZPP-02-143 (1 eq) in acetonitrile and add benzenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction mixture to room temperature. Filter, wash the filter cake with acetonitrile, and concentrate the filter cake under reduced pressure to obtain ZPP-02-149 (183 mg, yield 84.68%). LC-MS (ESI) C 19 H 19 N5O3 + [M+H] + Calculated value: 365.39; Measured value: 366.15.

[0225] Step 7: Referring to the method in step 3 of Example 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( SSynthesized the target product (19.00 mg, yield 24.53%) using LCMS (ESI) C-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridin-1-yl)isoindolin-2-yl)piperidine-2,6-dione. 32 H 29 ClN5O3 + [M+H] + Calculated value: 567.06; Measured value: 567.10. 1 H NMR (600 MHz, DMSO) δ 11.01 (s, 1H), 7.90 – 7.86 (m, 1H), 7.84 (d, J = 8.1 Hz, 1H), 7.72 (s, 1H), 7.65 (d, J = 7.6 Hz, 1H), 7.58 (dd, J = 8.1,1.7 Hz, 1H), 7.57 – 7.54 (m, 1H), 7.42 (qd, J = 6.9, 3.4 Hz, 3H), 7.37 – 7.32(m, 2H), 7.15 (dt, J = 6.3, 3.2 Hz, 1H), 5.14 (dd, J = 13.3, 5.1 Hz, 1H), 4.50 (d, J = 17.6 Hz, 1H), 4.39 (d, J = 17.6 Hz, 1H), 3.51 (d, J = 13.9 Hz, 1H), 3.42 (d, J = 13.9 Hz, 1H), 3.29 (s, 2H), 2.95 – 2.89 (m, 1H), 2.65 –2.58 (m, 4H), 2.54 (dd, J = 11.3, 5.3 Hz, 1H), 2.45 – 2.39 (m, 1H), 2.04 –2.00 (m, 1H).

[0226] Example 43: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-2-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-047)

[0227]

[0228] Step 1: Pyrazine-2-amine (1.0 eq) was dissolved in ethanol, and then 3-bromo-1,1,1-trifluoropropan-2-one (1.50 eq) was added. The reaction mixture was stirred at 80 °C for 12 h. The reaction was detected by LC-MS. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give LC-06-006 (7.13 g, yield 72.48%) as a white solid. LC-MS (ESI)C7H5F3N3 + [M+H] + Calculated value: 188.13; Measured value: 188.20.

[0229] Step 2: Dissolve LC-06-006 (1 eq) in dimethyl ethylene glycol and add palladium on carbon (10% W). Replace with hydrogen three times, and stir the reaction mixture at 60 °C for 12 h. After the reaction is complete, detect the reaction by LC-MS. Filter with diatomaceous earth, and evaporate the filtrate to dryness to obtain LC-06-025 (4.21 g, yield 82.42%). LC-MS (ESI) C7H9F3N3 + [M+H] + Calculated value: 192.16; Measured value: 192.20.

[0230] Step 3: Referring to the method in Step 3 of Example 27, 8-methyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine was replaced with 2-(trifluoromethyl)-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine to synthesize the target product. LCMS (ESI)C 12 H 17 F3N3O2 + [M+H] + Calculated value: 292.27; Measured value: 292.30.

[0231] Step 4: Referring to the method in Step 4 of Example 27, 8-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 12 H 16 BrF3N3O2 + [M+H] + Calculated values: 370.03 and 372.03; measured values: 370.10 and 372.10.

[0232] Step 5: Referring to the method in Step 3 of Example 1, 3-bromo-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 3-bromo-2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 29 H 37 F3N5O6 + [M+H] + Calculated value: 608.63; Measured value: 608.60.

[0233] Step 6: Referring to the method in Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS(ESI) C 20 H 19 F3N5O3 + [M+H] + Calculated value: 434.39; Measured value: 434.40.

[0234] Step 7: Referring to the method in Step 3 of Example 26, replace (S)-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione with (S)-3-(1-oxo-5-(2-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione to synthesize the target product. LCMS (ESI) C 33 H 28 ClF3N5O3 + [M+H] + Calculated value: 635.06; Measured value: 635.10. 1 H NMR (600 MHz, DMSO) δ 11.02 (s, 1H), 7.84 (d, J = 7.8 Hz, 1H), 7.70 (s, 1H), 7.64(d, J = 7.6 Hz, 1H), 7.56 (dd,J = 13.3, 5.6 Hz, 2H), 7.45 – 7.39 (m, 3H), 7.37 (td, J = 7.5, 0.9 Hz, 1H), 7.34 (dd, J = 7.2, 2.0 Hz, 1H), 7.17 (dd, J =7.5, 0.9 Hz, 1H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.53 (d, J = 17.6 Hz, 1H), 4.40 (d, J = 17.6 Hz, 1H), 3.70 (t, J = 5.4 Hz, 2H), 3.55 (t, J = 6.7 Hz, 3H), 3.45 (d, J = 13.9 Hz, 1H), 2.96 – 2.89 (m, 1H), 2.68 (ddd, J = 14.2,10.6, 4.5 Hz, 2H), 2.61 (d, J = 16.9 Hz, 1H), 2.45 – 2.39 (m, 1H), 2.03 (ddd, J = 10.2, 5.1, 3.1 Hz, 1H).

[0235] Example 44: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-2-methyl-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-06-052)

[0236]

[0237] Step 1: Following the method in Step 2 of Example 43, 2-(trifluoromethyl)imidazo[1,2-a]pyrazine was replaced with 6-bromo-2-methylimidazo[1,2-a]pyrazine to synthesize the target product. LCMS (ESI) C7H 12 N3 + [M+H] + Calculated value: 138.19; Measured value: 138.20.

[0238] Step 2: Referring to the method in Step 3 of Example 27, 8-methyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine was replaced with 2-methyl-5,6,7,8-tetrahydroimidazole[1,2-a]pyrazine to synthesize the target product. LCMS (ESI) C 12 H 20 N3O2 + [M+H] + Calculated value: 238.30; Measured value: 238.30.

[0239] Step 3: Referring to the method in Step 4 of Example 27, 8-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with 2-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 12 H 19 BrN3O2 + [M+H] + Calculated values: 316.06 and 318.06; measured values: 316.10 and 318.10.

[0240] Step 4: Referring to Step 3 of Example 1, replace 3-bromo-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester with 3-bromo-2-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 29 H 40 N5O6 + [M+H] + Calculated value: 554.66; Measured value: 554.60.

[0241] Step 5: Referring to Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-2-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI)C 20 H 22 N5O3 + [M+H] + Calculated value: 380.42; Measured value: 380.40.

[0242] Step 6: Referring to the method in step 3 of embodiment 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(5-(2-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione. LCMS (ESI) C 33 H 31 ClN5O3 + [M+H] + Calculated value: 581.09; Measured value: 581.10. 1 H NMR (600 MHz, DMSO) δ 11.02 (s, 1H), 7.86 (d, J = 7.8 Hz, 1H), 7.71 (s, 1H), 7.64 (d, J =7.5 Hz, 1H), 7.59 – 7.56 (m, 2H), 7.46 – 7.41 (m, 3H), 7.38 (d, J = 7.4 Hz, 1H), 7.35 (dd, J = 7.1, 2.0 Hz, 1H), 7.18 (d, J = 7.5 Hz, 1H), 5.15 (dd, J =13.3, 5.1 Hz, 1H), 4.52 (d, J = 17.5 Hz, 1H), 4.39 (s, 1H), 3.88 (t, J = 5.0Hz, 2H), 3.70 (s, 2H), 3.59 (d, J = 13.9 Hz, 1H), 3.51 – 3.48 (m, 1H), 2.95 –2.90 (m, 1H), 2.79 – 2.69 (m, 2H), 2.61 (ddd, J = 6.7, 5.6, 2.6 Hz, 1H), 2.43(dd, J = 13.1, 4.5 Hz, 1H), 2.22 (s, 3H), 2.04 – 2.01 (m, 1H).

[0243] Example 45: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-6-fluoro-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-06-065)

[0244]

[0245] Step 1: Following the method in Step 1 of Example 30, 4-bromo-2-ethylbenzoic acid was replaced with 4-bromo-5-fluoro-2-methylbenzoic acid to synthesize the target product. LCMS (ESI) C9H9BrFO2 + [M+H] + Calculated values: 246.97 and 248.97; measured values: 247.10 and 249.10.

[0246] Step 2: Dissolve LC-06-026 (1.0 eq) in 1,2-dichloroethane solution, then add NBS (2.0 eq) and AIBN (0.20 eq). Stir the reaction mixture at 85 °C for 12 h. Detect the completion of the reaction by TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-06-030 (4.60 g, yield 80.20%). LC-MS (ESI) C 10 H 11 Br2O2 + [M+H] + Calculated values: 404.79 and 406.79; measured values: 404.80 and 406.80.

[0247] Step 3: Dissolve LC-06-030 (1.0 eq) in tetrahydrofuran solution, then add diethyl phosphite (0.40 eq) and diisopropylethylamine (0.60 eq). Stir the reaction mixture at 25 °C for 12 h. Detect the reaction completion by TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-06-039 (587.00 mg, yield 88.91%). LC-MS (ESI) C9H8Br2FO2 + [M+H] + Calculated values: 324.88 and 326.88; measured values: 325.10 and 327.10.

[0248] Step 4: Referring to the method in Step 1 of Example 1, replace methyl 4-bromo-2-bromomethylbenzoate with methyl 4-bromo-2-(bromomethyl)-5-fluorobenzoate to synthesize the target product. LCMS (ESI) C 17 H 21 BrFN2O4 + [M+H] + Calculated value: 416.26; Measured value: 416.30.

[0249] Step 5: Referring to the method in Step 2 of Example 1, replace (S)-5-amino-4-(5-bromo-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester with (S)-5-amino-4-(5-bromo-6-fluoro-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester to synthesize the target product. LCMS (ESI) C 23 H 33 BFN2O6 + [M+H] + Calculated value: 463.33; Measured value: 463.30.

[0250] Step 6: Referring to the method in Step 1 of Example 26, tert-butyl (S)-5-amino-5-oxo-4-(1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)isoindololin-2-yl)valerate was replaced with (S)-5-amino-4-(6-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)isoindololin-2-yl)-5-oxovalerate to synthesize the target product. LCMS (ESI) C 27 H 36 FN6O6 + [M+H] + Calculated value: 559.61; Measured value: 559.60.

[0251] Step 7: Referring to Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-6-fluoro-1-oxoisoindoline-5-yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS(ESI) C 18 H 18 FN6O3 + [M+H] +Calculated value: 385.37; Measured value: 385.40.

[0252] Step 8: Referring to the method in step 3 of embodiment 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(6-fluoro-1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione. LCMS (ESI) C 31 H 27 ClFN6O3 + [M+H] + Calculated value: 586.04; Measured value: 586.10. 1 H NMR (600 MHz, DMSO) δ 11.03 (s, 1H), 7.88 (d, J = 6.0 Hz, 1H), 7.76 (d, J = 8.7Hz, 1H), 7.62 (d, J = 7.4 Hz, 1H), 7.56 – 7.53 (m, 1H), 7.45 – 7.37 (m, 4H), 7.34 (dd, J = 7.2, 2.0 Hz, 1H), 7.17 (dd, J = 7.5, 1.1 Hz, 1H), 5.16 (dd, J =13.3, 5.1 Hz, 1H), 4.52 (d, J = 17.4 Hz, 1H), 4.40 (d, J = 17.4 Hz, 1H), 3.86– 3.76 (m, 2H), 3.65 (s, 2H), 3.60 (d, J = 13.8 Hz, 1H), 3.48 (d, J = 13.8Hz, 1H), 2.92 (ddd, J = 17.5, 13.7, 5.4 Hz, 1H), 2.74 – 2.64 (m, 2H), 2.63 –2.58 (m, 1H), 2.45 – 2.39 (m, 1H), 2.06 – 2.01 (m, 1H).

[0253] Example 46: Synthesis of (S)-3-(5-(7-((2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-071)

[0254] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(3-methyl-1H-1,2,4-triazol-1-yl)nicotinaldehyde. LCMS (ESI) C 28 H 28 N9O3 + [M+H] + Calculated value: 538.59; Measured value: 538.60.

[0255] Example 47: Synthesis of (S)-3-(1-oxo-5-(7-(2-(2-oxopiridine-1-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidine-2,6-dione (LC-06-072)

[0256] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(2-oxopiperidin-1-yl)benzaldehyde. LCMS (ESI) C 31 H 33 N6O4 + [M+H] + Calculated value: 553.65; Measured value: 553.70. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.77 (d, J = 7.8Hz, 2H), 7.66 (d, J = 8.4 Hz, 1H), 7.54 (d, J = 7.1 Hz, 1H), 7.38 – 7.32 (m,2H), 7.27 – 7.22 (m, 2H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 (d, J = 17.3Hz, 1H), 4.36 (d, J= 17.3 Hz, 1H), 4.15 – 4.06 (m, 2H), 3.72 – 3.63 (m, 3H), 3.61 – 3.56 (m, 1H), 3.44 (t, J = 11.9 Hz, 2H), 2.94 – 2.89 (m, 1H), 2.88 –2.80 (m, 2H), 2.61 (dd, J = 9.8, 6.0 Hz, 1H), 2.43 – 2.34 (m, 3H), 2.04 –2.00 (m, 1H), 1.84 (qd, J = 14.1, 8.2 Hz, 4H).

[0257] Example 48: Synthesis of (S)-3-(1-oxo-5-(7-((2-oxo-2H-[1,2'-bipyridin]-3'-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidine-2,6-dione (LC-06-074)

[0258] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(1-methyl-2-oxo-1,2-dihydropyridin-3-yl)benzaldehyde. LCMS (ESI) C 30 H 28 N7O4 + [M+H] + Calculated value: 550.59; Measured value: 550.55. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 8.20 (dd, J = 4.7, 1.4 Hz, 1H), 8.17 (dd, J = 4.6, 1.5 Hz, 1H), 8.03 (d, J = 7.3Hz, 1H), 7.88 (dd, J = 10.7, 4.7 Hz, 1H), 7.79 – 7.73 (m, 2H), 7.65 (d, J =7.9 Hz, 1H), 7.29 (dd, J = 7.4, 4.9 Hz, 1H), 7.26 (s, 1H), 7.19 (dd, J= 6.9, 5.2 Hz, 1H), 7.13 (d, J = 8.2 Hz, 1H), 5.13 (dd, J = 13.4, 5.0 Hz, 1H), 4.48(d, J = 17.3 Hz, 1H), 4.36 (d, J = 17.3 Hz, 1H), 4.10 (t, J = 5.1 Hz, 2H), 3.82 (s, 2H), 3.78 (s, 2H), 2.91 (dd, J = 11.0, 6.2 Hz, 3H), 2.63 – 2.59 (m,1H), 2.42 (dd, J = 13.0, 4.4 Hz, 1H), 2.04 – 2.00 (m, 1H).

[0259] Example 49: Synthesis of (S)-3-(5-(7-((2'-bromo-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-082)

[0260] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-bromo-[1,1'-biphenyl]-2-carboxaldehyde. LCMS (ESI) C 32 H 29 BrN5O3 + [M+H] + Calculated value: 611.52; Measured value: 611.50. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.77 – 7.69 (m,3H), 7.65 (d, J = 7.5 Hz, 1H), 7.63 – 7.59 (m, 1H), 7.45 (dtd, J = 10.4, 7.6,1.2 Hz, 2H), 7.37 (td, J = 7.5, 1.1 Hz, 1H), 7.36 – 7.31 (m, 2H), 7.19 – 7.11(m, 2H), 5.13 (dd, J= 13.3, 5.1 Hz, 1H), 4.47 (d, J = 17.4 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.01 (t, J = 5.4 Hz, 2H), 3.51 (d, J = 16.7 Hz, 3H), 3.44(d, J = 13.8 Hz, 1H), 2.92 (ddd, J = 17.4, 13.7, 5.4 Hz, 1H), 2.76 – 2.64 (m,2H), 2.63 – 2.58 (m, 1H), 2.41 (tt, J = 11.0, 5.4 Hz, 1H), 2.01 (ddd, J =10.2, 5.2, 3.1 Hz, 1H).

[0261] Example 50: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-084)

[0262] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2'-chloro-[1,1'-biphenyl]-3-carboxaldehyde. LCMS (ESI) C 32 H 29 ClN5O3 + [M+H] + Calculated value: 567.07; Measured value: 567.10. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.75 (d, J = 7.4Hz, 2H), 7.65 (d, J = 8.3 Hz, 1H), 7.59 – 7.55 (m, 1H), 7.48 – 7.40 (m, 6H), 7.36 (d, J = 7.4 Hz, 1H), 7.18 (s, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.47(d,J = 17.3 Hz, 1H), 4.35 (d, J = 17.3 Hz, 1H), 4.11 (t, J = 5.2 Hz, 2H), 3.82 (s, 2H), 3.71 (s, 2H), 2.93 (dd, J = 13.6, 5.3 Hz, 1H), 2.90 (t, J = 5.3Hz, 2H), 2.62 – 2.58 (m, 1H), 2.41 (dt, J = 13.5, 8.9 Hz, 1H), 2.03 – 1.99(m, 1H).

[0263] Example 51: Synthesis of (S)-3-(5-(5-(3-(2-chlorophenyl)pyridinyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-098)

[0264] (S)-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)isoindololin-2-yl)piperidin-2,6-dione (1.00 eq) and 3-(2-chlorophenyl)pyridinecarboxylic acid (1.00 eq) were dissolved in DMF. HATU (1.50 eq) and DIPEA (5.00 eq) were added to the solution. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by thin-layer chromatography. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-TLC to give a white solid (S)-3-(5-(5-(3-(2-chlorophenyl)pyridinyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindololin-2-yl)piperidin-2,6-dione (21.00 mg, yield 26.44%). LCMS (ESI) C 30 H 25 ClN7O4 + [M+H] + Calculated value: 583.02; Measured value: 583.10.

[0265] Example 52: Synthesis of (S)-3-(5-(7-(3-(2-chlorophenyl)pyridinyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (LC-06-100)

[0266] Referring to the method of Example 51, the target product was synthesized by replacing (S)-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)isoindololin-2-yl)piperidine-2,6-dione with (S)-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione. LCMS(ESI) C 30 H 25 ClN7O4 + [M+H] + Calculated value: 583.02; Measured value: 583.10.

[0267] Example 53: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-4-fluoro-1-oxoisoindololin-2-yl)piperidine-2,6-dione (LC-06-121)

[0268]

[0269] Step 1: 4-Bromo-3-fluorobenzoic acid (1.0 eq) was dissolved in tetrahydrofuran, and the reaction system temperature was lowered to -78°C. Then, diisopropylaminolithium (2.1 eq) was added. The reaction solution was stirred at -78°C for 1 h. Subsequently, iodomethane (3.0 eq) was added dropwise to the reaction system, and the reaction solution was stirred at 20°C for another 12 h. The reaction was detected by LC-MS. The reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phase was washed with saturated sodium thiosulfate solution, saturated sodium bicarbonate solution, water, and saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid LC-06-088 (2.50 g, yield 46.99%). LC-MS (ESI) C8H7BrFO2 + [M+H] + Calculated values: 232.95 and 234.95; measured values: 233.10 and 235.10.

[0270] Step 2: Referring to the method in Step 1 of Example 30, 4-bromo-2-ethylbenzoic acid was replaced with 4-bromo-3-fluoro-2-methylbenzoic acid to synthesize the target product. LCMS (ESI) C9H9BrFO2 + [M+H] + Calculated values: 246.97 and 248.97; measured values: 247.10 and 249.10.

[0271] Step 3: Dissolve LC-06-091 (1.0 eq) in carbon tetrachloride solution, then add NBS (1.1 eq) and AIBN (0.20 eq). Stir the reaction mixture at 80 °C for 12 h. Detect the reaction completion by TLC. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-06-095 (6.13 g, yield 87.67%). LC-MS (ESI) C9H8Br2FO2 + [M+H] + Calculated values: 324.88 and 326.88; measured values: 325.10 and 327.10.

[0272] Step 4: Referring to the method in Step 1 of Example 1, replace methyl 4-bromo-2-bromomethylbenzoate with methyl 4-bromo-2-(bromomethyl)-3-fluorobenzoate to synthesize the target product. LCMS (ESI) C 17 H 21 BrFN2O4 + [M+H] + Calculated values: 415.06 and 417.06; measured values: 415.10 and 417.10.

[0273] Step 5: Referring to the method in Step 2 of Example 1, replace (S)-5-amino-4-(5-bromo-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester with (S)-5-amino-4-(5-bromo-4-fluoro-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester to synthesize the target product. LCMS (ESI) C 23 H 33 BFN2O6 + [M+H] + Calculated value: 463.33; Measured value: 463.30.

[0274] Step 6: Referring to the method in Step 1 of Example 26, tert-butyl (S)-5-amino-5-oxo-4-(1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)isoindololin-2-yl)valerate was replaced with tert-butyl (S)-5-amino-4-(4-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)isoindololin-2-yl)-5-oxovalerate to synthesize the target product. LCMS (ESI) C 27 H 36 FN6O6 + [M+H] + Calculated value: 559.61; Measured value: 559.60.

[0275] Step 7: Referring to the method in Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-5-amino-4-(4-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)isoindoline-2-yl)-5-oxopentanoic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 18 H 18 FN6O3 + [M+H] + Calculated value: 385.37; Measured value: 385.40.

[0276] Step 8: Referring to the method in step 3 of embodiment 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(4-fluoro-1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione. LCMS (ESI) C 31 H 27 ClFN6O3 + [M+H] + Calculated value: 586.04; Measured value: 586.10. 1 H NMR (600 MHz, DMSO) δ 11.04 (s, 1H), 7.81 – 7.75 (m, 1H), 7.73 (d, J= 7.7 Hz, 1H), 7.63 (d, J = 7.4 Hz, 1H), 7.56 – 7.53 (m, 1H), 7.45 – 7.37 (m, 4H), 7.34(dd, J = 7.0, 2.2 Hz, 1H), 7.17 (dd, J = 7.5, 1.1 Hz, 1H), 5.16 (dd, J =13.3, 5.1 Hz, 1H), 4.66 (d, J = 17.6 Hz, 1H), 4.50 (d, J = 17.6 Hz, 1H), 3.84(t, J = 5.1 Hz, 2H), 3.66 (s, 2H), 3.60 (d, J = 13.8 Hz, 1H), 3.48 (d, J =13.8 Hz, 1H), 2.95 – 2.90 (m, 1H), 2.73 – 2.65 (m, 2H), 2.62 (dd, J = 11.9,4.7 Hz, 1H), 2.48 – 2.45 (m, 1H), 2.04 (ddd, J = 10.2, 5.1, 3.1 Hz, 1H).

[0277] Example 54: Synthesis of (S)-3-(5-(5-(2'-chloro-[1,1'-biphenyl]-2-carbonyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-144)

[0278] Referring to the synthetic method of Example 51, the target product was synthesized by replacing 3-(2-chlorophenyl)pyridinecarboxylic acid with 2'-chloro-[1,1'-biphenyl]-2-carboxylic acid. LCMS (ESI) C 31 H 26 ClN6O4 + [M+H] + Calculated value: 582.03; Measured value: 582.10.

[0279] Example 55: Synthesis of (S)-3-(2-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-5-oxo-5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)piperidine-2,6-dione (LC-06-147)

[0280]

[0281] Step 1: Following the method in Step 1 of Example 30, 4-bromo-2-ethylbenzoic acid was replaced with 6-chloro-2-methylnicotinic acid to synthesize the target product. LCMS (ESI) C8H9ClNO2 + [M+H] + Calculated value: 186.61; Measured value: 186.60.

[0282] Step 2: Referring to the method in Step 3 of Example 53, methyl 4-bromo-3-fluoro-2-methylbenzoate was replaced with methyl 6-chloro-2-methylnicotinate to synthesize the target product. LCMS (ESI) C8H8BrClNO2 + [M+H] + Calculated values: 263.93 and 265.93; measured values: 264.10 and 265.10.

[0283] Step 3: Referring to the method in Step 1 of Example 1, methyl 4-bromo-2-bromomethylbenzoate was replaced with methyl 2-(bromomethyl)-6-chloronicotinic acid to synthesize the target product. LCMS (ESI) C 16 H 21 ClN3O4 + [M+H] + Calculated value: 354.80; Measured value: 354.50.

[0284] Step 4: Dissolve LC-06-094 (1 eq) and 1,1,1,2,2,2-hexabutyldistannane (1.5 eq) in 1,4-dioxane, and add lithium chloride (6 eq). Purge three times with nitrogen, then add Pd(dppf)Cl2 (0.05 eq) and PCy3 (0.10 eq), purging three times with nitrogen again. Heat the reaction mixture to 120 °C and stir for 12 h. Detect the reaction completion by LCMS, and cool the reaction mixture to room temperature. Add saturated potassium fluoride solution to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-06-104 (2.56 g, yield 74.43%). LC-MS (ESI) C 28 H48 N3O4Sn + [M+H] + Calculated value: 609.41; Measured value: 609.40.

[0285] Step 5: Dissolve LC-06-104 (1 eq) and 3-bromo-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) in 1,4-dioxane, and add lithium chloride (3 eq). Purge three times with nitrogen, then add Pd(PPh3)4Cl2 (0.05 eq) and CuI (1.50 eq), purging three times with nitrogen again. Heat the reaction mixture to 120 °C and stir for 12 h. Detect the reaction completion by LCMS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid LC-06-112 (635.00 mg, yield 71.33%). LC-MS (ESI) C 26 H 36 N7O6 + [M+H] + Calculated value: 542.61; Measured value: 542.60.

[0286] Step 6: Referring to the method in Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(6-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-5-oxo-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 17 H 18 N7O3 + [M+H] + Calculated value: 368.37; Measured value: 368.40.

[0287] Step 7: Referring to the method in step 3 of Example 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( SSynthesis of the target product using 3-(5-oxo-2-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)piperidine-2,6-dione. LCMS (ESI) C 30 H 27 ClN7O3 + [M+H] + Calculated value: 569.03; Measured value: 569.10. 1 H NMR (600 MHz, DMSO) δ 11.04 (s, 1H), 8.28 (s, 2H), 7.62 (d, J = 7.6Hz, 1H), 7.54 (dd, J = 5.3, 2.8 Hz, 1H), 7.45 – 7.42 (m, 1H), 7.42 – 7.37 (m,3H), 7.36 – 7.34 (m, 1H), 7.19 – 7.14 (m, 1H), 5.21 (dd, J = 13.3, 5.1 Hz, 1H), 4.58 (d, J = 17.8 Hz, 1H), 4.43 (d, J = 17.8 Hz, 1H), 4.36 (s, 2H), 3.66(s, 2H), 3.59 (dd, J = 13.7, 3.6 Hz, 1H), 3.47 (dd, J = 13.7, 2.8 Hz, 1H), 2.98 – 2.88 (m, 1H), 2.81 – 2.68 (m, 2H), 2.64 – 2.60 (m, 1H), 2.46 – 2.40(m, 1H), 2.09 – 2.03 (m, 1H).

[0288] Example 56: Synthesis of (S)-3-(5-(5-(2-(benzyloxy)benzyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-06-165)

[0289] Referring to step 4 of Example 36, the target product was synthesized by replacing 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(benzyloxy)benzaldehyde. LCMS (ESI) C 32 H31 N6O4 + [M+H] + Calculated value: 563.63; Measured value: 563.60. 1 HNMR (600 MHz, DMSO) δ 11.02 (s, 1H), 7.98 (d, J = 1.0 Hz, 1H), 7.92 (d, J =8.2 Hz, 1H), 7.84 (dd, J = 8.2, 1.7 Hz, 1H), 7.48 (d, J = 7.2 Hz, 2H), 7.41(dd, J = 7.5, 1.4 Hz, 1H), 7.37 (dd, J = 10.3, 4.7 Hz, 2H), 7.31 (dd, J =8.3, 6.3 Hz, 1H), 7.28 – 7.23 (m, 1H), 7.11 (d, J = 8.0 Hz, 1H), 6.96 (t, J =7.4 Hz, 1H), 5.18 – 5.13 (m, 3H), 4.56 (d, J = 17.7 Hz, 1H), 4.44 (d, J =17.7 Hz, 1H), 3.82 (s, 2H), 3.69 (s, 2H), 2.95 (t, J = 5.3 Hz, 2H), 2.93 –2.88 (m, 1H), 2.81 (t, J = 5.5 Hz, 2H), 2.64 – 2.59 (m, 1H), 2.43 (tt, J =13.3, 6.6 Hz, 1H), 2.04 (dtd, J = 12.3, 5.1, 2.0 Hz, 1H).

[0290] Example 57: Synthesis of (S)-3-(5-(5-([1,1'-biphenyl]-2-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione (LC-05-132)

[0291]

[0292] Step 1: LC-01-156 (1 eq) and 3-bromo-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylic acid tert-butyl ester (1 eq) were dissolved in 1,4-dioxane, and 2 M sodium carbonate solution (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(PPh3)4 (0.04 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-03-080 (520 mg, yield 43.33%). LC-MS (ESI) C 28 H 37 N5O6 + [M+H] + Calculated value: 539.63; Measured value: 540.28.

[0293] Step 2: (3 eq). The reaction mixture was heated to 100°C and stirred for 12 h. The reaction was monitored for completion by LC-MS, and the mixture was cooled to room temperature. The mixture was filtered, the filter cake was washed with acetonitrile, and the cake was concentrated under reduced pressure to obtain ZPP-03-082 (320 mg, yield 90.91%). LC-MS (ESI) C 19 H 19 N5O3 + [M+H] + Calculated value: 365.39; Measured value: 366.15.

[0294] Step 3: Referring to the method in step 5 of Example 1, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindolin-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(1-oxo-5-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione. LCMS (ESI)C 32 H 30 N5O3 + [M+H] + Calculated value: 532.62; Measured value: 532.60. 1 H NMR (600 MHz, DMSO) δ 10.99(s, 1H), 7.88 (s, 1H), 7.71 (d, J= 7.9 Hz, 1H), 7.60 (dd, J = 7.5, 1.0 Hz,1H), 7.55 (s, 1H), 7.47 – 7.43 (m, 3H), 7.37 (dddd, J = 10.8, 8.9, 6.1, 1.3Hz, 5H), 7.24 (dd, J = 7.4, 1.4 Hz, 1H), 5.12 (dd, J = 13.3, 5.1 Hz, 1H), 4.46 (d, J = 17.2 Hz, 1H), 4.34 (d, J = 17.1 Hz, 1H), 4.09 (t, J = 5.4 Hz,2H), 3.79 (s, 2H), 3.73 (s, 2H), 2.94 – 2.88 (m, 1H), 2.84 (t, J = 5.5 Hz, 2H), 2.61 (dd, J = 10.1, 6.4 Hz, 1H), 2.44 – 2.38 (m, 1H), 2.01 (ddd, J =10.3, 5.2, 3.1 Hz, 1H).

[0295] Example 58: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-149)

[0296]

[0297] Step 1: LC-01-156 (1 g, 1 eq) and 1-bromo-5,6-dihydroimidazolo[1,5-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) were dissolved in 1,4-dioxane, and 2 M sodium carbonate solution (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(PPh3)4 (0.04 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-03-088 (380 mg, yield 31.41%). LC-MS (ESI) C 28 H 37 N5O6 + [M+H] + Calculated value: 539.63; Measured value: 540.28.

[0298] ZPP-03-088 (1 eq) was dissolved in acetonitrile, and benzenesulfonic acid (3 eq) was added. The reaction mixture was heated to 100 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. The mixture was filtered, and the filter cake was washed with acetonitrile. The filter cake was concentrated under reduced pressure to obtain ZPP-03-091 (250 mg, yield 97.28%). LC-MS (ESI) C 19 H 19 N5O3 + [M+H] + Calculated value: 365.39; Measured value: 366.15.

[0299] Step 3: Referring to the method in step 3 of Example 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(1-oxo-5-(5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-1-yl)isoindololin-2-yl)piperidine-2,6-dione. LCMS(ESI) C 32 H 29 ClN5O3 + [M+H] + Calculated value: 567.06; Measured value: 567.10. 1 H NMR (600 MHz, DMSO) δ11.00 (s, 1H), 7.75 (d, J= 8.0 Hz, 1H), 7.71 (s, 1H), 7.62 (d, J = 7.3 Hz, 2H), 7.57 (dd, J = 6.0, 3.3 Hz, 1H), 7.45 – 7.37 (m, 4H), 7.37 – 7.31 (m,2H), 7.16 (d, J = 7.1 Hz, 1H), 5.12 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 (dd, J =17.3, 3.7 Hz, 1H), 4.35 (dd, J = 17.2, 3.4 Hz, 1H), 4.09 (s, 2H), 3.62 (dd, J = 96.8, 56.8 Hz, 4H), 2.95 – 2.89 (m, 1H), 2.74 (s, 2H), 2.63 – 2.59 (m, 1H), 2.43 – 2.38 (m, 1H), 2.02 (dtd, J = 7.1, 5.0, 2.0 Hz, 1H).

[0300] Example 59: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (LC-05-150)

[0301]

[0302] Step 1: LC-01-156 (1 g, 1 eq) and 3-iodo-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester (1 eq) were dissolved in 1,4-dioxane, and 2 M sodium carbonate solution (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(PPh3)4 (0.04 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, which was repeated three times until complete. The organic phase was washed with saturated brine, and the two phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-03-089 (300 mg, yield 25.67%). LC-MS (ESI) C28 H 37 N5O6 + [M+H] + Calculated value: 539.63; Measured value: 540.28.

[0303] Step 2: Dissolve ZPP-03-089 (1 eq) in acetonitrile and add benzenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction mixture to room temperature. Filter, wash the filter cake with acetonitrile, and concentrate the filter cake under reduced pressure to obtain ZPP-03-092 (180 mg, yield 88.67%). LC-MS (ESI) C 19 H 19 N5O3 + [M+H] + Calculated value: 365.39; Measured value: 366.15.

[0304] Step 3: Referring to the method in Step 3 of Example 26, (S)-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione was replaced with (S)-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoindololin-2-yl)piperidine-2,6-dione to synthesize the target product. LCMS (ESI) C 32 H 29 ClN5O3 + [M+H] + Calculated value: 567.06; Measured value: 567.10.

[0305] Example 60: Synthesis of (S)-3-(1-oxo-5-(7-(2-oxo-1,2-dihydropyridin-3-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidine-2,6-dione (XY-01-015)

[0306]

[0307] Step 1: 2-(benzyloxy)-3-bromopyridine (1 eq) and (2-formylphenyl)boronic acid (1.2 eq) were dissolved in dimethyl ethylene glycol and water, followed by the addition of sodium carbonate (2.5 eq) and tetrakis(triphenylphosphine)palladium (0.04 eq). Nitrogen was purged for 5 minutes, and the reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a pale yellow liquid, XY-01-008 (782.00 mg, yield 66.77%). LC-MS (ESI) C 19 H 16 NO2 + [M+H] + Calculated value: 290.33; Measured value: 290.54.

[0308] Step 2: Referring to the method in Step 5 of Example 1, replace 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(2-(benzyloxy)pyridin-3-yl)benzaldehyde to synthesize the target product. LCMS (ESI) C 38 H 35 N6O4 + [M+H] + Calculated value: 639.73; Measured value: 639.83.

[0309] Step 3: Dissolve XY-01-012 (1 eq) and palladium on carbon (0.1 eq.) in ethanol, displace hydrogen gas for 5 minutes, heat the reaction solution to 50°C, and stir for 24 h. Detect the completion of the reaction using LC-MS, and cool the reaction solution to room temperature. Add water to the reaction solution, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue using silica gel agar to obtain a white solid XY-01-015 (21.90 mg, yield 43.33%). LC-MS (ESI) C 31 H 29 N6O4 + [M+H] + Calculated value: 549.60; Measured value: 549.77. 1H NMR (600 MHz, ) δ 11.71 (s), 11.00 (s),7.76 (d, J = 7.9 Hz), 7.72 (s), 7.62 (d, J = 8.0 Hz), 7.57 (d, J = 7.6 Hz), 7.41 (dt, J = 6.2, 3.0 Hz), 7.38 – 7.34 (m), 7.31 (t, J = 7.4 Hz), 7.21 –7.15 (m), 6.26 (t, J = 6.6 Hz), 5.13 (dd, J = 13.3, 5.1 Hz), 4.48 (d, J =17.3 Hz), 4.36 (d, J = 17.3 Hz), 4.02 (t, J = 5.2 Hz), 3.64 (s), 3.57 (s), 2.96 – 2.88 (m), 2.73 (t, J = 5.1 Hz), 2.63 – 2.58 (m), 2.44 – 2.38 (m), 2.06– 1.97 (m).

[0310] Example 61: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-7-fluoro-1-oxoisoindol-2-yl)piperidine-2,6-dione (XY-01-051)

[0311]

[0312] Step 1: Methyl 4-bromo-2-fluoro-6-methylbenzoate (1 eq) and N-bromosuccinimide (3 eq) were dissolved in carbon tetrachloride, and then azobisisobutyronitrile (0.3 eq) was added. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS until completion, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain liquid XY-01-033 (5.11 g, yield 93.50%). LC-MS (ESI) C9H7Br3FO2 + [M+H] + Calculated values: 405.86 and 407.87; measured values: 405.88 and 405.90.

[0313] Step 2: XY-01-033 (1 eq) and diethyl phosphate (0.8 eq) were dissolved in tetrahydrofuran, and then N,N-diisopropylethylamine (1.2 eq) was added. The reaction mixture was stirred at room temperature for 12 h. The reaction was monitored by LC-MS to confirm completion, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid XY-01-038 (3.71 g, yield 85.05%). LC-MS (ESI) C9H8Br2FO2 + [M+H] + Calculated values: 326.96 and 328.97; Measured values: 326.97 and 328.99.

[0314] Step 3: Referring to the method in Step 1 of Example 1, methyl 4-bromo-2-bromomethylbenzoate was replaced with methyl 4-bromo-2-(bromomethyl)-6-fluorobenzoate to synthesize the target product. LCMS (ESI) C 17 H 21 BrFN2O4 + [M+H] + Calculated values: 416.26 and 418.30; measured values: 416.29 and 418.33.

[0315] Step 4: Referring to the method in Step 2 of Example 1, replace (S)-5-amino-4-(5-bromo-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester with (S)-5-amino-4-(5-bromo-7-fluoro-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester to synthesize the target product. LCMS (ESI) C 23 H 33 BFN2O6 + [M+H] + Calculated value: 463.33; Measured value: 463.36.

[0316] Step 5: Referring to the method in Step 1 of Example 26, tert-butyl (S)-5-amino-5-oxo-4-(1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)isoindololin-2-yl)valerate was replaced with (S)-5-amino-4-(7-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)isoindololin-2-yl)-5-oxovalerate was used to synthesize the target product. LCMS (ESI) C 27 H 36 FN6O6 + [M+H] +Calculated value: 558.61; Measured value: 559.69.

[0317] Step 6: Referring to Step 4 of Example 1, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxacyclopentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxacyclopentan-2-yl)-7-fluoro-1-oxoisoindoline-5-yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 18 H 18 FN6O3 + [M+H] + Calculated value: 385.37; Measured value: 385.20.

[0318] Step 7: Referring to the method in step 3 of Example 26, ( S )-3-(1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(7-fluoro-1-oxo-5-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)isoindol-2-yl)piperidine-2,6-dione. LCMS (ESI) C 31 H 27 ClFN6O3 + [M+H] + Calculated value: 586.06; Measured value: 586.10. 1H NMR (600MHz, ) δ 11.03 (s), 7.85 (s), 7.69 (d, J = 10.2 Hz), 7.64 (d, J = 7.6 Hz), 7.56 (dd, J = 6.5, 2.7 Hz), 7.45 (t, J = 7.3 Hz), 7.43 – 7.40 (m), 7.39 (d, J = 7.5 Hz), 7.36 (dd, J = 10.7, 4.2 Hz), 7.18 (d, J = 7.4 Hz), 5.11 (dd, J =13.3, 5.1 Hz), 4.55 (d, J = 18.0 Hz), 4.43 (d, J = 18.0 Hz), 4.14 (t, J = 5.5Hz), 3.66 (s), 3.57 (d, J = 13.8 Hz), 3.48 (d, J = 13.8 Hz), 2.95 – 2.87 (m), 2.76 – 2.66 (m), 2.61 (d, J = 16.8 Hz), 2.44 – 2.37 (m), 2.08 – 1.98 (m).

[0319] Example 62: Synthesis of (S)-3-(5-(7-((S)-2-(benzylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-3-carbonyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (XY-01-057)

[0320]

[0321] Step 1: (S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid methyl ester hydrochloride (1 eq) and benzyl sulfonyl chloride (1 eq) were dissolved in dichloromethane, and then triethylamine (2.1 eq) was added. The reaction mixture was stirred at room temperature for 20 h. The reaction was monitored by LC-MS to confirm completion, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a white solid XY-01-054 (920 mg, yield 72.50%). LC-MS (ESI) C 18 H 20 NO4S + [M+H] + Calculated value: 346.41; Measured value: 346.50.

[0322] Step 2: Dissolve XY-01-054 (1 eq) in a mixed solution of tetrahydrofuran, methanol, and water, and stir in an ice bath for 20 minutes. Add lithium hydroxide hydrate to the reaction solution, heat the reaction to 50°C, and stir for 12 h. Detect the completion of the reaction using LC-MS, and cool the reaction solution to room temperature. Add 1M hydrochloric acid to the reaction solution to adjust the pH to 6, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid XY-01-056 (600 mg, yield 81.25%). LC-MS (ESI) C 17 H 18 NO4S + [M+H] + Calculated value: 332.39; Measured value: 332.45.

[0323] Step 3: (S)-3-(1-oxo-5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidine-2,6-dione (1 eq) and XY-01-056 (1 eq) were dissolved in N,N-dimethylformimide, and N,N-diisopropylethylamine (3 eq) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 eq) were added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS to confirm completion. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate treatment to obtain XY-01-057 (30.00 mg, yield 33.54%) as a white solid. LC-MS(ESI) C 36 H 35 N6O6S + [M+H] + Calculated value: 679.76; Measured value: 679.80.

[0324] Example 63: Synthesis of (S)-N-(2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)phenyl)acetamide (XY-01-064)

[0325]

[0326] Step 1: Dissolve 2-aminobenzaldehyde (1 eq) and triethylamine (1.2 eq) in dichloromethane, then add acetyl chloride (1.5 eq). Stir the reaction mixture in an ice bath for 3 h. Detect the completion of the reaction by LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain liquid XY-01-061 (73.00 mg, yield 25.74%). LC-MS (ESI) C9H 10 NO2 + [M+H] + Calculated value: 164.18; Measured value: 164.25.

[0327] Step 2: Referring to the method in Step 5 of Example 1, replace 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with N-(2-formylphenyl)acetamide to synthesize the target product. LCMS (ESI) C 28 H 29 N6O4 + [M+H] + Calculated value: 513.17; Measured value: 513.23. 1 H NMR (600 MHz, ) δ 11.00 (s), 9.69 (s), 7.78 (d, J =7.9 Hz), 7.67 (d, J = 8.8 Hz), 7.39 (d, J = 7.4 Hz), 7.33 – 7.24 (m), 7.13(t, J = 7.4 Hz), 5.13 (dd, J = 13.3, 5.1 Hz), 4.49 (d, J = 17.4 Hz), 4.37 (d,J = 17.3 Hz), 4.15 (t, J = 5.2 Hz), 3.81 (s), 3.73 (s), 2.97 – 2.84 (m), 2.65– 2.57 (m), 2.42 (dd, J = 13.1, 4.6 Hz), 2.08 – 1.99 (m).

[0328] Example 64: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)sulfonyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (XY-01-102)

[0329]

[0330] Step 1: Dissolve (2-bromophenyl)methylamine (1 eq) and (2-chlorophenyl)boric acid (1.2 eq) in dimethyl ether and water, then add potassium carbonate (4 eq) and bis(triphenylphosphine)palladium dichloride (0.02 eq). Purge with nitrogen for 5 minutes, heat the reaction mixture to 80°C, and stir for 12 h. Detect the reaction completion by LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a pale yellow liquid, XY-01-087 (2.23 g, yield 76.78%). LC-MS (ESI) C 13 H 13 ClN + [M+H] + Calculated value: 217.70; Measured value: 217.89.

[0331] Step 2: XY-01-087 (1 eq), (1,4-diazabicyclo[2.2.2]octanedi(sulfur dioxide) adduct (0.6 eq), cuprous chloride (0.1 eq), and concentrated hydrochloric acid (4 eq) were dissolved in acetonitrile, and then tert-butyl nitrite (1.1 eq) was slowly added dropwise. The mixture was stirred at room temperature for 12 h. The reaction was monitored by LC-MS to confirm completion, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a pale yellow liquid XY-01-092 (198 mg, yield 36.33%). LC-MS (ESI) C 12 H9Cl2O2S + [M+H] + Calculated value: 288.15; Measured value: 288.35.

[0332] Step 3: Place ( S XY-01-092 (1 eq) was dissolved in dichloromethane, followed by the addition of 1 eq of 1-(1-oxo-5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidin-2,6-dione. The reaction mixture was stirred in an ice bath for 3 h. The reaction was monitored by LC-MS until complete, and the mixture was cooled to room temperature. Water was added to the reaction mixture, followed by extraction with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate analysis to give XY-01-102 (8.0 mg, 23.26% yield) as a white solid. LC-MS (ESI) C 31 H 27 ClN5O5S+ [M+H] + Calculated value: 617.09; Measured value: 617.32. 1 H NMR (600 MHz, ) δ11.00 (s), 8.13 (dd, J = 8.0, 0.9 Hz), 7.79 (ddd, J = 21.0, 10.5, 4.5 Hz), 7.73 – 7.68 (m), 7.60 (d, J = 7.7 Hz), 7.52 (d, J = 8.0 Hz), 7.48 – 7.42 (m), 7.41 – 7.35 (m), 7.20 (s), 5.13 (dd, J = 13.3, 5.1 Hz), 4.48 (d, J = 17.2Hz), 4.36 (d, J = 16.7 Hz), 4.16 (d, J = 15.9 Hz), 4.06 – 3.94 (m), 3.17 (d,J = 5.2 Hz), 2.92 (ddd,J = 17.5, 13.7, 5.4 Hz), 2.64 – 2.57 (m), 2.42 (ddd,J = 25.5, 12.8, 4.1 Hz), 2.09 – 1.89 (m).

[0333] Example 65: Synthesis of (S)-2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-N,N-dimethylbenzamide (XY-01-107)

[0334]

[0335] Step 1: Referring to Step 5 of Example 1, replace 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-formylbenzoic acid to synthesize the target product. LCMS (ESI) C 27 H 26 N5O5 + [M+H] + Calculated value: 500.53; Measured value: 500.58.

[0336] Step 2: XY-01-095 (1 eq) and a 2M tetrahydrofuran solution of dimethylamine (2 eq) were dissolved in N,N-dimethylformimide. N,N-diisopropylethylamine (5 eq) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 eq) were added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored for completion by LC-MS. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate analysis to obtain a white solid XY-01-107 (3.70 mg, yield 25.77%). LC-MS (ESI) C 29 H 31 N6O4+ [M+H]+, calculated value: 527.60; measured value: 527.80.

[0337] Example 66: Synthesis of (S)-2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-N-methylbenzamide (XY-01-108)

[0338] Referring to step 2 of Example 65, the tetrahydrofuran solution of dimethylamine was replaced with an aqueous solution of methylamine to synthesize the target product. LCMS (ESI) C 28 H 29 N6O4 + [M+H] + Calculated value: 513.57; Measured value: 513.66.

[0339] Example 67: Synthesis of (S)-3-(5-(6-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (XY-01-141)

[0340]

[0341] Step 1: Dissolve 3-bromo-1-methyl-pyrrolo[2,3-c]pyridine (1 eq) and benzyl bromide (1.2 eq) in acetonitrile, purge with nitrogen for 5 minutes, heat the reaction mixture to 80°C, and stir for 12 h. Detect the completion of the reaction using LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a colorless liquid LC-06-187 (900 mg, yield 73.26%). LC-MS (ESI) C 14 H 12 BrN3 + [M-Br] + Calculated values: 302.16 and 304.20; measured values: 302.29 and 304.31.

[0342] Step 2: Dissolve LC-06-187 (1 eq) and sodium borohydride (5 eq) in methanol and aqueous solution, purge with nitrogen for 5 minutes, heat the reaction mixture to 75°C, and stir for 12 h. Detect the completion of the reaction using LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a yellow liquid LC-06-196 (560 mg, yield 64.33%). LC-MS (ESI) C 14 H 17 BrN3 + [M+H] + Calculated values: 307.18 and 309.22; measured values: 307.22 and 309.37.

[0343] Step 3: Dissolve LC-06-196 (1 eq) and (S)-5-amino-5-oxo-4-(1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)isoindoline-2-yl)tert-butyl valerate (1 eq) in dioxane and water, then add potassium carbonate (2 eq) and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.05 eq). Purge with nitrogen for 5 minutes, heat the reaction mixture to 100°C, and stir for 12 h. Detect the completion of the reaction using LCMS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography to obtain a yellow liquid XY-01-129 (2.23 g, yield 76.78%). LC-MS (ESI) C 31 H38 N5O4 + [M+H] + Calculated value: 544.67; Measured value: 544.77.

[0344] Step 4: Refer to the method in step 4 of Example 1. LCMS (ESI) C 27 H 28 N5O3 + [M+H] + Calculated value: 470.55; Measured value: 470.59. ¹H NMR (600 MHz, ) δ 11.00 (s), 7.90 (s), 7.83 (d, J = 8.5 Hz), 7.76 (s), 7.42 (s), 5.12 (dd, J = 13.3, 5.1 Hz), 4.48 (d, J = 17.3 Hz), 4.36 (d, J = 17.2 Hz), 3.73 (s), 2.92 (ddd, J = 17.5, 13.7, 5.4 Hz), 2.63 – 2.57 (m), 2.40 (dt, J = 13.3, 8.8 Hz), 2.15 – 1.74 (m).

[0345] Step 5: Dissolve XY-01-134 (1 eq) and palladium on carbon (0.1 eq.) in methanol, purge with hydrogen for 5 minutes, heat the reaction mixture to 50°C, and stir for 24 h. Detect the completion of the reaction using LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid XY-01-136 (132 mg, yield 33.23%). LC-MS (ESI) C 20 H 22 N5O3 + [M+H] + Calculated value: 380.42; Measured value: 380.55.

[0346] Step 6: Referring to the method in step 3 of embodiment 26, ( S )-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentane-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester is replaced with ( SSynthesis of the target product using 1-(5-(1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione. LCMS (ESI) C 33 H 31 ClN5O3 + [M+H] + Calculated value: 581.08; Measured value: 581.10. ¹H NMR (600 MHz, ) δ 10.99 (s), 7.85 (s), 7.79 (d, J = 8.0 Hz), 7.72 (d, J = 8.0 Hz), 7.64 (d, J = 7.7 Hz), 7.55 (dd, J = 5.9, 3.3 Hz), 7.47 – 7.39 (m), 7.38 – 7.33 (m), 7.16 (d, J = 7.5 Hz), 5.12 (dd, J = 13.3, 5.1 Hz), 4.47 (d, J = 17.2 Hz), 4.35 (d, J = 17.2 Hz), 3.65 (s), 3.5 ... = 14.0 Hz), 3.44 (dd, J = 19.4, 13.3 Hz), 2.96 – 2.87 (m), 2.78 – 2.53(m), 2.40 (qd, J = 13.1, 4.4 Hz), 2.18 – 1.84 (m).

[0347] Example 68: Synthesis of (S)-3-(5-(6-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (XY-01-147)

[0348]

[0349] Step 1: Referring to the method in Step 1 of Example 67, 3-bromo-1-methyl-pyrrolo[2,3-c]pyridine was replaced with 3-bromo-1H-pyrrolo[2,3-c]pyridine to synthesize the target product. LC-MS (ESI) C 14 H 10 BrN3 + [M-Br] + Calculated values: 288.16 and 290.21; measured values: 288.22 and 290.33.

[0350] Step 2: Referring to the method in Step 2 of Example 67, 6-benzyl-3-bromo-1-methyl-pyrazolo[3,4-c]pyridine-6-onium was replaced with 6-benzyl-3-bromo-1H-pyrazolo[3,4-c]pyridine-6-onium to synthesize the target product. LC-MS (ESI)C 13 H 15 BrN3 + [M+H] + Calculated values: 293.18 and 295.22; measured values: 293.25 and 295.44.

[0351] Step 3: Referring to the method in Step 3 of Example 67, 6-benzyl-3-bromo-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine was replaced with 6-benzyl-3-bromo-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine to synthesize the target product. LC-MS (ESI) C 30 H 36 N5O4 + [M+H] + Calculated value: 530.64; Measured value: 530.77.

[0352] Step 4: Refer to the method in step 4 of Example 1. LC-MS (ESI) C 26 H 26 N5O3 + [M+H] + Calculated value: 456.52; Measured value: 456.66.

[0353] Step 5: Referring to the method in Step 5 of Example 67, (S)-3-(5-(6-benzyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione was replaced with (S)-3-(5-(6-benzyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione to synthesize the target product. LC-MS (ESI) C 19 H 20 N5O3 + [M+H] + Calculated value: 366.39; Measured value: 366.52.

[0354] Step 6: Referring to the method in Step 3 of Example 26, (S)-3-(2-(1-amino-5-(tert-butoxy)-1,5-dioxapentan-2-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester was replaced with (S)-3-(1-oxo-5-(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)isoindoline-2-yl)piperidine-2,6-dione to synthesize the target product. LCMS (ESI) C 32 H 29 ClN5O3 + [M+H] + Calculated value: 567.06; Measured value: 567.33.

[0355] Example 69: Synthesis of (S)-3-(1-oxo-5-(7-(benzenesulfonyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindolin-2-yl)piperidine-2,6-dione (XY-01-149)

[0356] Referring to step 3 of Example 64, the target product was synthesized by replacing 2'-chloro-[1,1'-biphenyl]-2-sulfonyl chloride with benzenesulfonyl chloride. LCMS (ESI) C 25 H 24 N5O5S + [M+H] + Calculated value: 506.55; Measured value: 506.76. ¹H NMR (600 MHz) δ 11.00 (s), 8.02 – 7.84 (m), 7.85 – 7.71 (m), 7.71 – 7.61 (m), 7.57 (d, J = 8.8 Hz), 7.18 (s), 5.13 (dd, J = 13.3, 5.1 Hz), 4.48 (d, J = 17.4 Hz), 4.35 (t, J = 8.6 Hz), 4.11 (t, J = 5.5 Hz), 3.54 (t, J = 5.5 Hz), 3.00 – 2.79 (m), 2.68 – 2.55 (m), 2.44 – 2.36 (m), 2.01 (ddd, J = 9.8, 5.0, 2.4 Hz).

[0357] Example 70: Synthesis of (S)-3-(5-(6-benzyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (XY-01-150)

[0358]

[0359] Step 1: Following the method of Step 1 in Example 67, 3-bromo-1-methyl-pyrrolo[2,3-c]pyridine was replaced with 3-bromo-1-methyl-1H-pyrrolo[2,3-c]pyridine to synthesize the target product. LC-MS (ESI) C 15 H 13 BrN2 + [M-Br] + Calculated values: 301.19 and 303.21; measured values: 301.22 and 303.43.

[0360] Step 2: Referring to the method in Step 2 of Example 67, 6-benzyl-3-bromo-1-methyl-pyrazolo[3,4-c]pyridine-6-onium was replaced with 6-benzyl-3-bromo-1H-pyrazolo[3,4-c]pyridine-6-onium to synthesize the target product. LC-MS (ESI)C 15 H 18 BrN2 + [M+H] + Calculated values: 305.22 and 307.28; measured values: 305.25 and 307.48.

[0361] Step 3: Referring to the method in Step 3 of Example 67, 6-benzyl-3-bromo-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine was replaced with 6-benzyl-3-bromo-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridine to synthesize the target product. LC-MS (ESI) C 32 H 39 N4O4 + [M+H] + Calculated value: 543.68; Measured value: 543.87.

[0362] Step 4: Refer to the method in step 4 of Example 1. LC-MS (ESI) C 28 H 29 N4O3 + [M+H] + Calculated value: 469.56; Measured value: 469.76.

[0363] Example 71: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-02-171)

[0364]

[0365] Step 1: Dissolve tert-butyl 4-oxoperidin-1-carboxylate (1 eq) in anhydrous tetrahydrofuran, cool to -70°C with dry ice, and add lithium diisopropylamino (2 M in THF, 1.05 eq) to the solution at low temperature, stirring for 1 h. Add ethyl trifluoroacetate to the solution, and stir the reaction system at room temperature for 3 h. The reaction is confirmed by LCMS. Quench the reaction with saturated ammonium chloride solution, adjust the pH of the solution to neutral, extract with ethyl acetate, and concentrate the combined organic layers under reduced pressure. The residue is purified by silica gel column chromatography to obtain ZPP-03-026. LCMS (ESI) C 12 H 16 F3NO4 + [M+H] + Calculated value: 295.26; Measured value: 296.11.

[0366] Step 2: Synthesis of methyl 2-(dibromomethyl)-4-nitrobenzene (ZPP-02-097)

[0367] Methyl 2-methyl-4-nitrobenzene (10 g, 1 eq), N-bromosuccinimide (1.5 eq), and azobisisobutyronitrile (0.2 eq) were dissolved in carbon tetrachloride. The reaction mixture was heated to 85 °C and stirred for 12 h. The reaction was monitored by LC-MS until completion, and the mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give ZPP-02-097 (13.8 g, 85.71% yield). LC-MS (ESI) C9H7Br2NO4 + [M+H] + Calculated value: 352.97; Measured value: 353.88.

[0368] Step 3: ZPP-02-097 (1 eq) and diethyl phosphate (0.8 eq) were dissolved in tetrahydrofuran, and then N,N-diisopropylethylamine (1.2 eq) was added. The reaction mixture was stirred at room temperature for 12 h. The reaction was monitored by LC-MS to confirm completion. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-02-099 (10 g, yield 93.46%). LC-MS (ESI) C9H8BrNO4 + [M+H] + Calculated value: 274.07; Measured value: 275.96.

[0369] Step 4: ZPP-02-099 (1 eq) and (S)-4,5-diamino-5-oxovalerate tert-butyl hydrochloride (1.3 eq) were dissolved in acetonitrile, and then N,N-diisopropylethylamine (3 eq) was added. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-02-100 (12 g, yield 86.33%). LC-MS (ESI) C 17 H 21 N3O6 + [M+H] + Calculated value: 363.37; Measured value: 364.15.

[0370] Step 5: Dissolve ZPP-02-100 (1 eq) in methanol and add palladium on carbon (10% W). Replace with hydrogen three times, and stir the reaction mixture at room temperature for 12 h. After the reaction is complete, detect the reaction by LC-MS. Filter with diatomaceous earth, and evaporate the filtrate to dryness to obtain ZPP-02-106 (10.80 g, yield 98.18%). LC-MS (ESI) C 17 H 23 N3O4 + [M+H] + Calculated value: 333.39; Measured value: 334.17.

[0371] Step 6: Dissolve ZPP-02-106 (1 eq) in acetonitrile and add benzenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction mixture to room temperature. Filter, wash the filter cake with acetonitrile, and concentrate the filter cake under reduced pressure to obtain ZPP-02-107 (7.5 g, yield 89.28%). LC-MS (ESI) C 13 H 13 N3O3 + [M+H] + Calculated value: 259.27; Measured value: 260.10.

[0372] Step 7: Dissolve ZPP-02-107 (1 eq) in 6M HCl, and slowly add an aqueous solution of sodium nitrite (1 eq) dropwise at 0℃. Stir the reaction mixture at 0℃ for 1 h. Dissolve SnCl2·H2O in concentrated hydrochloric acid, and slowly add it dropwise to the reaction mixture at 0℃, continuing the reaction for 2 h. Detect the completion of the reaction using LC-MS, and adjust the pH to 7 by adding sodium bicarbonate solution. Filter, wash the filter cake with water, evaporate the filtrate to dryness, filter again, wash the filter cake with ethanol, and concentrate the filtrate under reduced pressure to obtain ZPP-02-150 (6.5 g, yield 81.97%). LC-MS (ESI) C 13 H 14 N4O3 + [M+H] + Calculated value: 274.28; Measured value: 275.11.

[0373] Step 8: Dissolve ZPP-02-150 (1 eq) in ethanol, add ZPP-03-026, purge with nitrogen three times, heat the reaction solution to 80℃, and stir for 12 h. Detect the completion of the reaction using LC-MS, and cool the reaction solution to room temperature. Add water to the reaction solution, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-02-157 (170 mg, yield 21.85%). LC-MS (ESI) C 25 H 26 F3N5O5 + [M+H] + Calculated value: 533.51; Measured value: 534.19.

[0374] Step 9: Dissolve ZPP-02-157 (1 eq) in dichloromethane and add 4M ethyl acetate hydrochloride solution. Stir the reaction mixture at room temperature for 2 h. The reaction was monitored by LC-MS to confirm completion. The reaction mixture was concentrated under reduced pressure to obtain ZPP-02-170 (126 mg, yield 91.32%). LC-MS (ESI) C 20 H 18 F3N5O3 + [M+H] +Calculated value: 433.39; Measured value: 434.14. ¹H NMR (600 MHz, DMSO) δ 11.03 (s, 1H), 9.90 (s, 2H), 7.92 (d, J = 8.1 Hz, 1H), 7.81 (s, 1H), 7.63 (d, J = 8.0 Hz, 1H), 5.16 (dd, J = 13.3, 5.0 Hz, 1H), 4.57 (d, J = 17.8 Hz, 1H), 4.44 (d, J = 17.8 Hz, 1H), 4.39 (s, 2H), 3.46 (t, J = 5.9 Hz, 2H), 3.06 (t, J = 5.8 Hz, 2H). 2.97 – 2.85 (m, 1H), 2.61 (d, J = 16.8 Hz, 1H), 2.41 (qd, J = 13.1, 4.1 Hz, 1H), 2.06 (dd, J = 11.2, 5.5 Hz, 1H).

[0375] Step 10: ZPP-02-170 (1 eq) and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) were dissolved in N,N-dimethylformimide, and a catalytic amount of acetic acid was added. The reaction mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium triacetoxyborohydride (3 eq), and stirring was continued for 12 h. The reaction was detected by LC-MS to indicate completion, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate to obtain a white solid ZPP-02-171 (16.00 mg, yield 15.69%). LC-MS (ESI) C 33 H 27 ClF3N5O3 + [M+H]+, calculated value: 634.06; measured value: 635.17. 1H NMR (400 MHz, DMSO) δ 11.03 (s, 1H), 7.88 (d, J = 8.0Hz, 1H), 7.74 (s, 1H), 7.65 – 7.52 (m, 3H), 7.38 (tdd, J = 21.9, 15.0, 6.7Hz, 5H), 7.16 (d, J = 7.4 Hz, 1H), 5.14 (dd, J = 13.2, 5.0 Hz, 1H), 4.55 (d,J = 17.8 Hz, 1H), 4.41 (d, J = 17.9 Hz, 1H), 3.55 (d, J = 13.8 Hz, 1H), 3.45(d, J = 14.8 Hz, 3H), 2.89 (s, 2H), 2.73 (s, 1H), 2.65 (d, J = 7.4 Hz, 3H), 2.40 (dd, J = 13.0, 4.5 Hz, 1H), 2.10 – 1.99 (m, 1H).

[0376] Example 72: Synthesis of (S)-3-(5-(5-([1,1'-biphenyl]-2-ylmethyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (ZPP-02-172)

[0377] Referring to step 10 of Example 71, 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde was replaced with [1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 33 H 28 F3N5O3 + [M+H] + Calculated value: 599.61; Measured value: 600.22. 1H NMR (400 MHz, DMSO) δ 10.93 (s, 1H), 7.90 (t, J = 15.6 Hz, 1H), 7.75 (s, 1H), 7.71 – 7.54 (m, 2H), 7.55 – 7.28 (m, 7H), 7.25 (d, J = 7.1 Hz,1H), 5.14 (d, J = 8.2 Hz, 1H), 4.46 (dt, J = 53.6, 16.2 Hz, 2H), 3.61 (d, J =46.2 Hz, 2H), 3.50 (d, J = 17.2 Hz, 2H), 2.96 (dd, J = 50.6, 37.6 Hz, 2H),2.66 (dd, J = 43.7, 13.1 Hz, 4H), 2.46 – 2.35 (m, 1H), 2.03 (s, 1H).

[0378] Example 73: Synthesis of (S)-3-(5-(5-benzyl-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-168)

[0379] Referring to step 10 of Example 71, 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde was replaced with benzaldehyde to synthesize the target product. LCMS (ESI) C 27 H 24 F3N5O3 + [M+H] + Calculated value: 523.52; Measured value: 524.19. 1H NMR (600MHz, DMSO) δ 11.02 (s, 1H), 7.89 (d, J = 8.1 Hz, 1H), 7.77 (s, 1H), 7.61 (d,J = 8.0 Hz, 1H), 7.39 – 7.34 (m, 4H), 7.28 (dd, J = 9.2, 4.3 Hz, 1H), 5.15(dd, J = 13.3, 5.1 Hz, 1H), 4.55 (d, J = 17.8 Hz, 1H), 4.42 (d, J = 17.8 Hz,1H), 3.75 (s, 2H), 3.64 (s, 2H), 2.92 (ddd, J = 17.8, 13.9, 5.5 Hz, 1H), 2.79(dd, J = 8.6, 3.9 Hz, 4H), 2.63 – 2.59 (m, 1H), 2.45 – 2.36 (m, 1H), 2.07 –2.01 (m, 1H).

[0380] Example 74: Synthesis of (3S)-3-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4-methyl-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-037)

[0381]

[0382] Step 1: Following the method of Step 1 in Example 71, replace 4-oxopiperidin-1-carboxylic acid tert-butyl ester with 2-methyl-4-oxopiperidin-1-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 13 H 18 F3NO4 + [M+H] + Calculated value: 309.29; Measured value: 310.12.

[0383] Step 2: Referring to Step 8 of Example 71, tert-butyl 4-oxo-3-(2,2,2-trifluoroacetyl)piperidine-1-carboxylate was replaced with tert-butyl 2-methyl-4-oxo-3-(2,2,2-trifluoroacetyl)piperidine-1-carboxylate to synthesize the target product (130 mg, yield 83.27%). LCMS (ESI) C 26 H 28 F3N5O5 + [M+H] +Calculated value: 547.54; Measured value: 548.21.

[0384] Step 3: Referring to the method in step 9 of Example 71, replace (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-(trifluoromethyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester with 1-(2-(( S Synthesis of the target product: 2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-4-methyl-3-(trifluoromethyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. LCMS (ESI)C 21 H 20 F3N5O3 + [M+H] + Calculated value: 447.42; Measured value: 448.16. 1 H NMR (600 MHz, DMSO) δ 11.02(s, 1H), 7.94 – 7.90 (m, 2H), 7.75 (d, J = 7.7 Hz, 1H), 5.15 (dd, J = 13.2,4.8 Hz, 1H), 4.57 (t, J = 15.2 Hz, 1H), 4.44 (d, J = 17.6 Hz, 1H), 3.58 (s,1H), 3.24 – 3.20 (m, 1H), 3.08 (dd, J = 16.4, 10.0 Hz, 1H), 2.93 (dd, J =20.7, 7.6 Hz, 1H), 2.87 (dd, J = 30.5, 16.9 Hz, 1H), 2.62 (d, J = 15.1 Hz, 1H), 2.55 (dd, J = 21.1, 9.6 Hz, 1H), 2.46-2.34 (m, 2H), 2.07 – 2.02 (m, 1H), 1.43 (d, J = 6.2 Hz, 3H).

[0385] Step 4: ZPP-03-036 (1 eq) and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) were dissolved in N,N-dimethylformamide, and a catalytic amount of acetic acid was added. The reaction mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium triacetoxyborohydride (3 eq), and stirring was continued for 12 h. The reaction was detected by LC-MS to indicate completion, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate to obtain a white solid ZPP-03-037 (6.00 mg, yield 14.83%). LC-MS (ESI) C 34 H 29 ClF3N5O3 + [M+H]+, calculated value: 648.08; measured value: 649.19. 1 H NMR (600 MHz, DMSO) δ 11.02 (s, 1H), 7.87 (dd, J = 12.5, 7.1 Hz,2H), 7.75 – 7.71 (m, 1H), 7.56 – 7.52 (m, 2H), 7.46 – 7.33 (m, 5H), 7.32 –7.28 (m, 1H), 7.15 (d, J = 6.6 Hz, 1H), 5.15 (dd, J = 13.3, 5.1 Hz, 1H), 4.54(d, J = 17.5 Hz, 1H), 4.42 (d, J = 17.7 Hz, 1H), 3.55 (dd, J = 13.6, 10.0 Hz,1H), 3.49 – 3.46 (m, 1H), 3.45 (d, J = 4.7 Hz, 1H), 3.04 – 2.96 (m, 1H), 2.96– 2.91 (m, 2H), 2.90 (d, J = 6.3 Hz, 1H), 2.65 – 2.58 (m, 2H), 2.47 (d, J =5.6 Hz, 1H), 2.43 (dd, J = 13.2, 4.4 Hz, 1H), 2.03 (dd, J = 8.8, 3.7 Hz, 1H), 1.24 (s, 3H).

[0386] Example 75: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-02-186)

[0387]

[0388] Step 1: Following the method in Step 1 of Example 80, benzoyl chloride was replaced with acetyl chloride to synthesize the target product. LCMS (ESI) C 12 H 19 NO4 + [M+H] + Calculated value: 241.29; Measured value: 242.13.

[0389] Step 2: Referring to the method in Step 8 of Example 71, 4-oxo-3-(2,2,2-trifluoroacetyl)piperidine-1-carboxylic acid tert-butyl ester was replaced with 3-acetyl-4-oxopiperidine-1-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI)C 25 H 29 N5O5 + [M+H] + Calculated value: 479.54; Measured value: 480.22.

[0390] Step 3: Referring to Step 9 of Example 71, (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-(trifluoromethyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester was replaced with (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-methyl-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 20 H 21 N5O3 + [M+H] + Calculated value: 379.42; Measured value: 380.17.

[0391] Step 4: Referring to the method in step 10 of embodiment 71, ( S )-3-(5-(5-(((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione replaced with (S Synthesis of the target product using 3-(5-(3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione. LCMS (ESI) C 33 H 30 ClN5O3 + [M+H] + Calculated value: 580.08; Measured value: 581.20. 1 H NMR (400 MHz, DMSO) δ 11.02 (s, 1H), 7.88 (t, J = 24.3 Hz, 1H), 7.69 (d, J = 32.0 Hz, 1H), 7.56 (t, J = 25.8 Hz, 4H), 7.44 (s, 3H), 7.38-7.30 (m, 1H), 7.22 (d, J = 52.2 Hz, 1H), 7.14 – 5.84 (m,1H), 5.28 – 5.01 (m, 1H), 4.46 (dd, J = 51.2, 17.6 Hz, 2H), 4.17 (dd, J =55.0, 16.7 Hz, 1H), 3.62 (s, 1H), 3.25 – 3.16 (m, 1H), 2.93 (dd, J = 22.6,9.6 Hz, 2H), 2.73 (s, 1H), 2.61 (d, J = 16.2 Hz, 1H), 2.52 (d, J = 13.8 Hz,1H), 2.45 – 2.37 (m, 1H), 2.22 (s, 3H), 2.07 – 1.99 (m, 1H), 1.93 (d, J =14.9 Hz, 1H), 1.31-1.12 (m, 1H).

[0392] Example 76: Synthesis of (S)-3-(5-(5-([1,1'-biphenyl]-2-ylmethyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (ZPP-02-187)

[0393] Referring to step 4 of Example 75, 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde was replaced with [1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 33 H 31 N5O3 +[M+H] + Calculated value: 545.64; Measured value: 546.38.

[0394] Example 77: Synthesis of (S)-3-(5-(3-benzyl-5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-011)

[0395]

[0396] Step 1: Following the method in Step 1 of Example 79, benzoyl chloride was replaced with 2-phenylacetyl chloride to synthesize the target product. LCMS (ESI) C 18 H 23 NO4 + [M+H] + Calculated value: 317.38; Measured value: 318.17.

[0397] Step 2: Referring to the method in Step 8 of Example 71, 4-oxo-3-(2,2,2-trifluoroacetyl)piperidine-1-carboxylic acid tert-butyl ester was replaced with 4-oxo-3-(2-phenylacetyl)piperidine-1-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI)C 31 H 33 N5O5 + [M+H] + Calculated value: 555.63; Measured value: 556.25.

[0398] Step 3: Referring to Step 9 of Example 71, (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-(trifluoromethyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester was replaced with (S)-3-(5-(3-benzyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione to synthesize the target product. LCMS (ESI) C 26 H 25 N5O3 + [M+H] + Calculated value: 455.52; Measured value: 456.20.

[0399] Step 4: Referring to the method of step 10 in Example 71, replace (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione with ( S Synthesis of the target product using 3-(5-(3-benzyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione. LCMS (ESI) C 39 H 34 ClN5O3 + [M+H] + Calculated value: 656.18; Measured value: 657.23. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.74 (t, J = 11.0 Hz, 1H), 7.62 (s, 1H), 7.56 – 7.48 (m, 3H), 7.37 (ddd, J = 24.5,15.5, 7.3 Hz, 4H), 7.27 (d, J = 7.2 Hz, 1H), 7.21 (t, J = 7.4 Hz, 2H), 7.13 (dt, J = 30.6, 13.0 Hz, 2H), 6.94 (d, J = 7.4 Hz, 2H), 5.12 (dd, J = 13.3,5.1 Hz, 1H), 4.41 (dd, J = 73.9, 17.5 Hz, 2H), 4.01 (s, 2H), 3.45 (d, J =41.7 Hz, 1H), 3.32 – 3.19 (m, 1H), 3.00 (d, J = 49.3 Hz, 2H), 2.95–2.87 (m,1H), 2.69 – 2.52 (m, 5H), 2.40 (qd, J = 13.2, 4.3 Hz, 1H), 2.04 – 1.96 (m,1H).

[0400] Example 78: Synthesis of (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-1-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-017)

[0401]

[0402] Step 1: 1-Bromo-5,6-dihydroimidazolo[1,5-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) was dissolved in 1,4-dioxane, and potassium carbonate (3 eq) and 2,4,6-trimethyl-1,3,5,2,4,6-tricycloribosane were added. The mixture was purged with nitrogen three times, and tetrakis(triphenylphosphine)palladium was added, followed by another nitrogen purging. The reaction mixture was heated to 110 °C and stirred for 6 h. The reaction was monitored by LC-MS to confirm completion, and the mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-03-003 (470 mg, yield 85.45%). LC-MS (ESI) C 12 H 19 N3O2 + [M+H] + Calculated value: 237.30; Measured value: 238.15.

[0403] Step 2: Dissolve ZPP-03-003 (1 eq) in acetonitrile, and add N-iodosuccinimide (1 eq). Stir the reaction mixture at room temperature for 16 h. Detect the completion of the reaction by LC-MS. Add saturated sodium thiosulfate solution to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-03-007 (650 mg, yield 83.27%). LC-MS (ESI) C 12 H 18 IN3O2 + [M+H] + Calculated value: 363.20; Measured value: 364.05.

[0404] Step 3: Dissolve ZPP-03-007 (1 eq) in 1,4-dioxane, add 2M sodium carbonate solution and tert-butyl 5-amino-5-oxo-4-(1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)isoindololin-2-yl)valerate. Purge with nitrogen three times, add tetrakis(triphenylphosphine)palladium, purge with nitrogen again, heat the reaction solution to 85°C, and stir for 12 h. Detect the reaction completion by LC-MS, and cool the reaction solution to room temperature. Add water to the reaction solution, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-03-008 (230 mg, yield 23.23%). LC-MS (ESI) C 29 H 39 N5O6+ [M+H] + Calculated value: 553.66; Measured value: 554.29.

[0405] Step 4: Dissolve ZPP-03-008 (1 eq) in acetonitrile and add benzenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction mixture to room temperature. Filter, wash the filter cake with acetonitrile, and concentrate the filter cake under reduced pressure to obtain ZPP-03-016 (50 mg, yield 31.81%). LC-MS (ESI) C 20 H 21 N5O3 + [M+H] + Calculated value: 379.42; Measured value: 380.17.

[0406] Step 5: ZPP-03-016 (1 eq) and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) were dissolved in N,N-dimethylformimide, and a catalytic amount of acetic acid was added. The reaction mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium triacetoxyborohydride (3 eq), and stirring was continued for 12 h. The reaction was detected by LC-MS to indicate completion, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate to obtain ZPP-03-017 (4.00 mg, yield 5.22%). LC-MS (ESI) C 33 H 30 ClN5O3 + [M+H] + Calculated value: 580.08; Measured value: 581.20.

[0407] Example 79: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-phenyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-115)

[0408]

[0409] Step 1: Dissolve tert-butyl 4-oxopiperidinium-1-carboxylate (2 g, 1 eq) in anhydrous tetrahydrofuran, cool to -70°C with dry ice, and add 10 mL of bis(trimethylsilylamino)lithium to the solution at low temperature, stirring for 1 h. Add benzoyl chloride to the solution, and stir the reaction system at room temperature for 3 h. The reaction is confirmed by LCMS. Quench the reaction with saturated ammonium chloride solution, adjust the pH of the solution to neutral, extract with ethyl acetate, and concentrate the combined organic layers under reduced pressure. The residue is purified by silica gel column chromatography to obtain ZPP-03-077. LCMS (ESI) C 17 H 21 NO4 + [M+H] + Calculated value: 303.36; Measured value: 304.15.

[0410] Step 2: Referring to Step 8 of Example 71, tert-butyl 4-oxo-3-(2,2,2-trifluoroacetyl)piperidine-1-carboxylic acid was replaced with tert-butyl 3-benzoyl-4-oxopiperidine-1-carboxylic acid to synthesize the target product (100 mg, yield 11.28%). LCMS (ESI) C 30 H 31 N5O5 + [M+H] + Calculated value: 541.61; Measured value: 542.24.

[0411] Step 3: Referring to Step 9 of Example 71, (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-(trifluoromethyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester was replaced with (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-phenyl-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 25 H 23 N5O3 + [M+H] + Calculated value: 441.49; Measured value: 442.18.

[0412] Step 4: Place ZPP-03-113 ( S3-(1-oxo-5-(3-phenyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)isoindololin-2-yl)piperidin-2,6-dione (1 eq) and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) were dissolved in N,N-dimethylformamide, and a catalytic amount of acetic acid was added. The reaction mixture was stirred at room temperature for 30 min, followed by the addition of sodium triacetoxyborohydride (3 eq), and stirring was continued for 12 h. The reaction was detected by LCMS to indicate completion, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate to give a white solid ZPP-03-115 (13.00 mg, yield 22.41%). LC-MS (ESI) C 38 H 32 ClN5O3 + [M+H] + Calculated value: 642.16; Measured value: 643.22. 1 H NMR (600 MHz, DMSO) δ 10.99 (s, 1H), 7.62 (d, J =8.2 Hz, 1H), 7.59 (d, J = 7.7 Hz, 1H), 7.55 (d, J = 7.4 Hz, 2H), 7.38 (ddd, J= 18.3, 9.1, 5.2 Hz, 6H), 7.31 (d, J = 7.3 Hz, 1H), 7.28 – 7.25 (m, 1H), 7.17(d, J = 8.3 Hz, 1H), 7.10 (d, J = 6.7 Hz, 3H), 5.09 (dd, J = 13.3, 5.1 Hz,1H), 4.41 (d, J = 17.6 Hz, 1H), 4.28 (d, J = 17.5 Hz, 1H), 3.51 – 3.40 (m,2H), 3.30 (s, 2H), 2.93 – 2.86 (m, 1H), 2.63 (dd, J = 51.1, 25.4 Hz, 5H), 2.37 (qd, J = 13.2, 4.4 Hz, 1H), 2.02-1.97 (m, 1H).

[0413] Example 80: Synthesis of (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-cyclopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-082)

[0414]

[0415] Step 1: Dissolve tert-butyl 4-oxopiperidinium-1-carboxylate (1 eq) in anhydrous tetrahydrofuran, cool to 0°C with ice, and add diisopropylaminolithium (2 M in THF, 1.05 eq) to the solution at low temperature, stirring for 1 h. Add cyclopropylyl chloride to the solution, and stir the reaction system at room temperature for 3 h. The reaction is confirmed by LCMS. Quench the reaction with saturated ammonium chloride solution, adjust the pH of the solution to neutral, extract with ethyl acetate, and concentrate the combined organic layers under reduced pressure. The residue is purified by silica gel column chromatography to obtain ZP-03-040. LCMS (ESI) C 14 H 21 NO4 + [M+H] + Calculated value: 267.32; Measured value: 268.15.

[0416] Step 2: Referring to the method in Step 8 of Example 71, tert-butyl 4-oxo-3-(2,2,2-trifluoroacetyl)piperidine-1-carboxylic acid was replaced with tert-butyl 3-(cyclopropanecarbonyl)-4-oxopiperidine-1-carboxylic acid to synthesize the target product. LCMS (ESI)C 27 H 31 N5O5 + [M+H] + Calculated value: 505.57; Measured value: 506.24.

[0417] Step 3: Referring to Step 9 of Example 71, (S)-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-3-(trifluoromethyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester was replaced with (S)-3-cyclopropyl-1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester to synthesize the target product. LCMS (ESI) C 22 H 23 N5O3 + [M+H] + Calculated value: 405.46; Measured value: 406.18.

[0418] Step 4: Referring to the method in step 10 of embodiment 71, ( S )-3-(5-(5-(((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione replaced with ( S Synthesis of the target product using 3-(5-(3-cyclopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione. LCMS (ESI) C 35 H 32 ClN5O3 + [M+H] + Calculated value: 606.12; Measured value: 607.22. 1 H NMR (600 MHz, DMSO) δ 11.01 (s, 1H), 7.84 (s, 1H), 7.81 – 7.78 (m, 1H), 7.78 – 7.74 (m, 1H), 7.63 (d, J = 7.6 Hz, 1H), 7.58 – 7.54 (m, 1H), 7.45 – 7.40 (m, 3H), 7.38 – 7.34 (m, 1H), 7.34 – 7.32(m, 1H), 7.15 (dd, J = 7.5, 0.9 Hz, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H),4.51 (d, J = 17.5 Hz, 1H), 4.38 (d, J = 17.5 Hz, 1H), 3.49 (d, J = 14.2 Hz, 1H), 3.42 (d, J = 13.9 Hz, 1H), 3.27 (dt, J = 15.7, 6.5 Hz, 2H), 2.92 (ddd, J= 17.5, 13.7, 5.5 Hz, 1H), 2.63 – 2.53 (m, 5H), 2.45 – 2.37 (m, 1H), 2.03(ddd, J = 10.2, 5.1, 3.1 Hz, 1H), 1.94 (td, J = 8.3, 4.2 Hz, 1H), 0.80 (dd, J= 8.4, 1.6 Hz, 2H), 0.41-0.32 (m, 2H).

[0419] Example 81: Synthesis of (S)-3-(5-(6-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione (ZPP-03-128)

[0420]

[0421] Step 1: 4,7-Dihydrothiopheno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (1 eq) was dissolved in dichloromethane, and N-bromosuccinimide (3 eq) was added. The reaction mixture was stirred at room temperature for 16 h. The reaction was monitored by LC-MS to confirm completion. Saturated sodium thiosulfate solution was added, and the mixture was extracted three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain ZPP-03-083 (1.4 g, yield 84.33%). LC-MS (ESI) C 12 H 15 Br2NO2S + [M+H] + Calculated value: 397.12; Measured value: 398.91.

[0422] Step 2: Dissolve ZPP-03-083 (1 eq) in anhydrous tetrahydrofuran, purging with nitrogen for 5 minutes. Cool to -78°C with dry ice, and add n-butyllithium (1.05 eq) to the solution at low temperature, stirring for 3 h. LC-MS detection indicates completion. Quench the reaction with saturated ammonium chloride solution, adjust the pH to neutral, extract with ethyl acetate, and concentrate the combined organic layers under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-03-105 (600 mg, yield 54.54%). LC-MS (ESI) C 12 H 16 BrNO2S + [M+H] + Calculated value: 318.23; Measured value: 319.01.

[0423] Step 3: Dissolve ZPP-03-105 (1 eq) in 1,4-dioxane, add 2M sodium carbonate solution and tert-butyl 5-amino-5-oxo-4-(1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)isoindolin-2-yl)valerate. Purge with nitrogen three times, add tetra(triphenylphosphine)palladium, purge with nitrogen again, heat the reaction mixture to 85°C, and stir for 12 h. Detect the reaction completion by LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain ZPP-03-108 (255 mg, yield 25.01%). LC-MS (ESI) C 13 H 13 N3O3 + [M+H] + Calculated value: 555.69; Measured value: 556.24.

[0424] Step 4: Dissolve ZPP-03-108 (1 eq) in acetonitrile and add benzenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction mixture to room temperature. Filter, wash the filter cake with acetonitrile, and concentrate the filter cake under reduced pressure to obtain ZPP-03-120 (100 mg, yield 58.49%). LC-MS (ESI) C 20 H 19 N3O3S + [M+H] + Calculated value: 381.45; Measured value: 382.12.

[0425] Step 5: ZPP-03-120 (1 eq) and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) were dissolved in N,N-dimethylformamide, and a catalytic amount of acetic acid was added. The reaction mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium triacetoxyborohydride (3 eq), and stirring was continued for 12 h. The reaction was detected by LC-MS to indicate completion, and sodium bicarbonate solution was added to quench the reaction mixture. Ethyl acetate was added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate to obtain ZPP-03-128 (24.00 mg, yield 31.57%). LC-MS (ESI) C 33 H 28 ClN3O3S + [M+H ]+ Calculated value: 582.12; Measured value: 583.15.

[0426] Example 82: Synthesis of (S)-3-(5-(7-(2-(1-methyl-2-oxo-1,2-dihydropyridin-3-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione (LC-06-073)

[0427] Referring to step 5 of Example 1, the target product was synthesized by replacing 2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde with 2-(1-methyl-2-oxo-1,2-dihydropyridin-3-yl)benzaldehyde. LCMS (ESI) C 32 H 31 N6O4 + [M+H] + Calculated value: 563.63; Measured value: 563.70. 1 H NMR (600 MHz, DMSO) δ 11.00 (s, 1H), 7.76(d, J = 7.9 Hz, 1H), 7.72 (dd, J = 5.6, 2.7 Hz, 2H), 7.62 (d, J = 8.0 Hz, 1H), 7.56 (d, J = 7.5 Hz, 1H), 7.37 (ddd, J = 15.9, 7.2, 1.6 Hz, 2H), 7.30(dd, J = 10.7, 4.2 Hz, 1H), 7.22 – 7.12 (m, 2H), 6.28 (t, J = 6.8 Hz, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 (d, J = 17.4 Hz, 1H), 4.36 (d, J = 17.3Hz, 1H), 4.00 (t, J = 5.1 Hz, 2H), 3.61 (s, 2H), 3.55 (s, 2H), 3.45 (s, 3H), 2.95 – 2.89 (m, 1H), 2.71 (t, J = 5.0 Hz, 2H), 2.63 – 2.59 (m, 1H), 2.44 –2.39 (m, 1H), 2.02 (dd, J= 8.9, 3.6 Hz, 1H).

[0428] Example 83: Synthesis of 3-{6-[7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl]-1-benzofuran-3-yl}hexahydropyridine-2,6-dione (LTL-03-086)

[0429]

[0430] Step 1: At room temperature, compound 1-(4-bromo-2-hydroxyphenyl)ethyl-1-one (1.0 eq) was dissolved in a mixed solvent of chloroform and ethyl acetate, and then copper bromide (2.0 eq) was added. The reaction mixture was heated to 100 °C and stirred for 14 h. The reaction was confirmed by LC-MS. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with dichloromethane. The filtrate was collected and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain LTL-02-077 (29.5 g, yield 86.5%). LC-MS (ESI) C8H6Br2O2 + [M+H] + Calculated value: 294.87; Measured value: 294.99.

[0431] Step 2: Under nitrogen protection, LTL-02-077 (1.0 eq) was dissolved in dichloromethane. The reaction solution was cooled to 0°C, and triethylamine (1.5 eq) was slowly added dropwise. The reaction mixture was slowly heated to 20°C and stirred for 30 minutes. After the reaction was complete as detected by LC-MS, water was added to the system, the mixture was separated, the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove about half of the solvent. Toluene was then added, and the mixture was further concentrated under reduced pressure to remove the remaining low-boiling solvent, yielding LTL-02-079, which was directly used in the next step. LC-MS (ESI) C8H5BrO2 + [M+H] + Calculated values: 212.95 and 214.95; measured values: 212.88 and 214.9.

[0432] Step 3: Under nitrogen protection and at room temperature, LTL-02-079 (1.0 eq) was dissolved in toluene, and tert-butyloxycarbonylmethylenetriphenylphosphine (1.0 eq) was added. The reaction mixture was heated to 130 °C and stirred for 20 h. After the reaction was complete as detected by LC-MS, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography to obtain LTL-02-193 (15.26 g). LC-MS (ESI) C 14 H 15 BrO3 + [M+H] +Calculated value: 311.02; Measured value: 311.23.

[0433] Step 4: Dissolve LTL-02-193 (1 eq) and pinacol diborate (1.5 eq) in 1,4-dioxane, and add potassium acetate (3 eq). Purge three times with nitrogen, add Pd(dppf)Cl2 (0.05 eq), and purge three more times with nitrogen. Heat the reaction mixture to 100°C and stir for 12 h. Detect the reaction completion by LC-MS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain LTL-02-196 (11.83 g). LC-MS (ESI) C 20 H 27 BO5 + [M+H] + Calculated value: 359.20; Measured value: 359.51.

[0434] Step 5: Dissolve LTL-02-196 (1.0 eq) in N,N-dimethylformamide, add potassium carbonate solid (1.0 eq), and add benzyltriethylammonium chloride (1.0 eq) dropwise while stirring. After stirring the reaction mixture at room temperature for 30 minutes, add acrylonitrile (1.0 eq). After the addition is complete, stir the reaction mixture at room temperature for 16 h. The reaction is confirmed by LC-MS. Dilute the reaction mixture with water and extract with ethyl acetate. Dry the organic layer with anhydrous sodium sulfate and concentrate under reduced pressure. The residue is purified by silica gel column chromatography to obtain LTL-02-200 (6.38 g, yield 62%). LC-MS (ESI) C 23 H 30 BNO 5+ [M+H] + Calculated value: 412.22; Measured value: 412.38.

[0435] Step 6: LTL-02-200 (1 eq) and 3-bromo-5,6-dihydroimidozop[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) were dissolved in 1,4-dioxane, and 2 M cesium carbonate solution (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(PPh3)4 (0.05 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 90 °C and stirred for 12 h. The reaction was monitored by LC-MS until complete. The reaction mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction. The extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid, LTL-03-073 (435 mg, yield 60.96%). LC-MS (ESI) C 28 H 34 N4O5 + [M+H] + Calculated value: 506.25; Measured value: 506.55.

[0436] Step 7: Dissolve LTL-03-073 (1 eq) in dimethyl sulfoxide and add potassium carbonate solid (0.2 eq). While stirring, add 70% hydrogen peroxide solution (0.5 eq) dropwise and stir at room temperature for 10 minutes. After the reaction is complete as detected by LC-MS, add water and ethyl acetate to the reaction solution for extraction. Repeat the extraction three times until complete. Wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain crude LTL-03-078, which can be used directly in the next step. LC-MS (ESI) C 28 H 36 N4O6 + [M+H] + Calculated value: 524.62; Measured value: 524.78.

[0437] Step 8: Dissolve LTL-03-078 (1 eq) in acetonitrile and add p-toluenesulfonic acid (3 eq). Heat the reaction mixture to 100°C and stir for 4 h. Detect the completion of the reaction using LCMS. Cool the reaction mixture to room temperature. Remove the solvent by vacuum concentration. Add water and dichloromethane to the residue. Adjust the pH of the aqueous phase to 7-8 with saturated sodium carbonate solution. Repeat extraction with dichloromethane until complete. Wash the organic phase with saturated brine. Combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under vacuum. Purify the residue using a pre-prepared silica gel plate to obtain a white solid LTL-03-085 (125 mg, 75% yield). LCMS (ESI) C 19 H 18 N4O3 + [M+H] +Calculated value: 351.14; Measured value: 351.44. 1 H NMR (600 MHz, DMSO-d6) δ 10.93 (s, 1H),7.97 (s, 1H), 7.70 (d, J = 1.4 Hz, 1H), 7.66 (d, J = 8.1 Hz, 1H), 7.36 (dd, J= 8.1, 1.5 Hz, 1H), 7.16 (s, 1H), 4.17 (d, J = 11.6 Hz, 3H), 4.12 (t, J = 5.5Hz, 2H), 2.76 (ddd, J = 17.5, 12.3, 5.4 Hz, 2H), 2.64 – 2.55 (m, 2H), 2.34(td, J = 12.6, 4.4 Hz, 2H), 2.14 (ddd, J = 11.5, 5.2, 2.5 Hz, 1H).

[0438] Step 9: Dissolve LTL-03-085 (1.0 eq) and 1-naphthaldehyde (1.2 eq) in N,N-dimethylformamide, and add a catalytic amount of acetic acid. Stir the reaction mixture at room temperature for 30 minutes, then add sodium cyanoborohydride (3.0 eq) and continue stirring for 3 hours. Detect the completion of the reaction using LCMS, and quench the reaction mixture with sodium bicarbonate solution. Extract the reaction mixture with ethyl acetate, repeating the extraction three times until complete. Wash the organic phase with saturated brine, combine the organic phases, dry them with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue using a pre-prepared silica gel plate to obtain a white solid, LTL-03-086 (11.00 mg, yield 43.99%). LCMS (ESI) C 30 H 26 N4O3 + [M+H] +Calculated value: 491.20; Measured value: 491.33. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.93 (s, 1H), 8.34 (dd, J = 7.8, 1.8 Hz, 1H), 7.97 –7.88 (m, 3H), 7.70 (d, J = 1.4 Hz, 1H), 7.60 (d, J = 8.1 Hz, 1H), 7.57 – 7.49 (m, 4H), 7.36 (dd, J = 8.1, 1.4 Hz, 1H), 7.05 (s, 1H), 4.15 (q, J = 6.6, 5.7Hz, 3H), 4.04 (t, J = 5.4 Hz, 2H). 3.74 (s, 2H), 2.93 (t, J = 5.4 Hz, 2H), 2.75 (ddd, J = 17.5, 12.4, 5.4 Hz, 1H), 2.58 (dt, J = 17.3, 4.0 Hz, 1H), 2.34(qd, J = 12.6, 4.4 Hz, 1H), 2.11 (dq, J = 13.5, 4.9 Hz, 1H).

[0439] Example 84: Synthesis of 3-(6-{7-[(2-phenylphenyl)methyl]-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl}-1-benzofuran-3-yl)hexahydropyridine-2,6-dione (LTL-03-087)

[0440] Referring to step 8 of Example 83, 1-naphthaldehyde was replaced with [1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 32 H 28 N4O3 + [M+H] +Calculated value: 517.22; Measured value: 517.34. ¹H NMR (600MHz, DMSO-d6) δ 10.94 (s, 1H), 7.95 (s, 1H), 7.73–7.58 (m, 3H), 7.49–7.25 (m, 9H), 7.05 (s, 1H), 4.16 (dd, J = 12.2, 4.9 Hz, 1H), 4.03 (t, J = 5.5 Hz, 2H), 3.65 (s, 2H), 3.58 (s, 2H), 2.75 (h, J = 6.3, 5.4 Hz, 3H), 2.62–2.56 (m, 1H), 2.35 (dd, J = 12.6, 4.3 Hz, 2.35 Hz, 2.62–2.56 (m, 1H), 2.35 (dd, J = 12.6, 4.3 Hz, 2.49–7.25 (m, 9H), 7.05 (s, 1H), 4.16 (dd, J = 12.2, 4.9 Hz, 1H), 4.03 (t, J = 5.5 Hz, 2H), 2.35 (dd, J = 12.6, 4.3 Hz, 2.49–7.58 (m, 3H), 2.35 (dd, J = 12.6, 4.3 Hz, 2.49–7.25 (m, 9H), 2.49–7.25 (m, 9H), 2.49–7.25 (m, 9H), 2.49–7.25 (m, 9H), Hz, 1H), 2.16–2.09 (m, 1H).

[0441] Example 85: Synthesis of 3-{5-[7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl]-1-benzofuran-3-yl}hexahydropyridine-2,6-dione (LTL-03-030)

[0442] Referring to step 8 of Example 83, the target product was synthesized by replacing 3-(6-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)benzofuran-3-yl)piperidine-2,6-dione with 3-(5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)benzofuran-3-yl)piperidine-2,6-dione. LCMS (ESI) C 30 H 26 N4O3 + [M+H] +Calculated value: 491.20; Measured value: 491.33. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.88 (s, 1H), 8.33 (dd, J = 8.0, 1.7 Hz, 1H), 8.01 (d, J = 2.7 Hz, 1H), 7.96 (dd, J = 7.4, 2.0 Hz, 1H), 7.92 (d, J = 8.2 Hz, 1H), 7.83 (d, J = 1.8 Hz, 1H), 7.73 (d, J = 8.6 Hz, 1H), 7.56 (ddd, J = 9.4, 7.3, 1.7 Hz, 4H), 7.53 – 7.49 (m, 2H), 4.23 (s, 2H), 4.17 (dd, J = 12.2, 4.8 Hz, 1H), 4.11 (q, J = 5.5 Hz, 2H), 4.02 (s, 2H), 3.03 (d,J = 5.7 Hz, 2H), 2.74 – 2.70 (m, 1H), 2.59 – 2.55 (m, 1H), 2.38 (dd, J =12.5, 4.4 Hz, 1H), 2.10 (ddd, J = 11.2, 5.1, 2.5 Hz, 1H).

[0443] Example 86: Synthesis of 3-(5-{7-[(2-phenylphenyl)methyl]-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl}-1-benzofuran-3-yl)hexahydropyridine-2,6-dione (LTL-03-049)

[0444] Referring to step 8 of Example 83, 3-(6-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)benzofuran-3-yl)piperidin-2,6-dione was replaced with 3-(5-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)benzofuran-3-yl)piperidin-2,6-dione, and 1-naphthaldehyde was replaced with [1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 32 H 28 N4O3 + [M+H] +Calculated value: 517.22; Measured value: 517.29. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.89 (s, 1H), 7.95 (s, 1H), 7.69 (d, J = 1.8 Hz, 1H), 7.65 – 7.60 (m, 2H), 7.45 – 7.36 (m, 8H), 7.27 (dd, J = 7.4, 1.6 Hz, 1H), 6.98 (s, 1H), 4.18 (dd, J = 12.1, 4.8 Hz, 1H), 3.95 (t, J = 5.4 Hz, 2H), 3.64 (s, 2H), 3.58 (s, 2H), 2.77 – 2.69 (m, 3H), 2.58 (dt, J = 17.3, 4.0 Hz, 1H), 2.38 (qd, J =12.5, 4.5 Hz, 1H), 2.13 – 2.09 (m, 1H).

[0445] Example 87: Synthesis of 3-{5-[5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl]-1-benzofuran-3-yl}hexahydropyridine-2,6-dione (LTL-03-080)

[0446] Referring to step 8 of Example 83, the target product was synthesized by replacing 3-(6-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)benzofuran-3-yl)piperidine-2,6-dione with 3-(5-(4,5,6,7-tetrahydropyrazin-3-yl)benzofuran-3-yl)piperidine-2,6-dione. LCMS (ESI) C 30 H 26 N4O3 + [M+H] +Calculated value: 491.20; Measured value: 491.13. ¹H NMR (600 MHz, DMSO-d6) δ 10.94 (s, 1H), 8.30 – 8.27 (m, 1H), 7.89 (d, J = 6.9 Hz, 2H), 7.85 (d, J = 8.2 Hz, 1H), 7.75 (s, 1H), 7.56 (d, J = 8.5 Hz, 1H), 7.53 (dd, J = 7.1, 1.3 Hz, 1H), 7.51 – 7.48 (m, 3H), 7.46 (dd, J = 8.2, 6.9 Hz, 1H), 7.28 (dd, J = 8.5, 1.8 Hz, 1H). 4.22 – 4.16 (m, 2H), 4.15 – 4.12 (m, 1H), 4.10 (t, J = 5.6 Hz, 2H), 3.91 (s, 2H), 3.03 (q, J = 5.7 Hz, 2H), 2.76 (ddd, J = 17.3, 12.0, 5.3 Hz, 1H), 2.61 (dt, J= 17.2, 4.2 Hz, 1H), 2.38 – 2.30 (m, 1H), 2.15 – 2.11 (m, 1H).

[0447] Example 88: Synthesis of 3-(5-{5-[(2-phenylphenyl)methyl]-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl}-1-benzofuran-3-yl)hexahydropyridine-2,6-dione (LTL-03-082)

[0448] Referring to step 8 of Example 83, 3-(6-(5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)benzofuran-3-yl)piperidin-2,6-dione was replaced with 3-(5-(4,5,6,7-tetrahydropyrazinzo[1,5-a]pyrazin-3-yl)benzofuran-3-yl)piperidin-2,6-dione, and 1-naphthaldehyde was replaced with [1,1'-biphenyl]-2-carboxaldehyde to synthesize the target product. LCMS (ESI) C 32 H 28 N4O3 + [M+H] +Calculated value: 517.22; Measured value: 517.39. ¹H NMR (600 MHz, DMSO-d6) δ 10.91 (s, 1H), 7.90 (s, 1H), 7.73 (s, 1H), 7.58 (dd, J = 7.8, 5.3 Hz, 2H), 7.51 (d, J = 1.7 Hz, 1H), 7.43 (t, J = 7.5 Hz, 2H), 7.40 – 7.31 (m, 5H), 7.23 (ddd, J = 10.4, 7.9, 1.7 Hz, 2H), 4.15 (dd, J = 11.9, 4.9 Hz, 1H), 4.07 (t, J = 5.6 Hz, 2H). 3.72 (d, J = 21.0 Hz, 4H), 2.84 (t, J = 5.7 Hz, 2H), 2.74 (td, J = 12.1, 6.0 Hz, 1H), 2.59 (dt, J = 17.3, 4.2 Hz, 1H), 2.35 (qd, J= 12.2, 4.3 Hz, 1H), 2.15 – 2.10 (m, 1H).

[0449] Example 89: Synthesis of 3-{2-chloro-3-[7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl]phenyl}hexahydropyridine-2,6-dione (LTL-03-179)

[0450]

[0451] Step 1: 3-Bromo-5,6,7,8-tetrahydroimidazolo[1,2-a]pyrazine (1.0 eq) and 1-(bromomethyl)naphthalene (1.1 eq) were dissolved in N,N-dimethylformamide, and triethylamine (3.0 eq) was added. The reaction mixture was stirred at room temperature for 3 h. The reaction was monitored by LC-MS to confirm completion. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a white solid, LTL-03-065 (90 g, yield 90.6%). LC-MS (ESI) C 17 H 16 BrN3 + [M+H] + Calculated values: 342.05 and 344.05; measured values: 342.34 and 344.28.

[0452] Step 2: At room temperature, 1-bromo-2-chloro-3-toluene (1.0 eq) was dissolved in carbon tetrachloride, followed by the addition of benzoyl peroxide (0.08 eq). The reaction mixture was heated to 100 °C and stirred for 16 h. N-bromosuccinimide was added in portions. The reaction was monitored by LC-MS to indicate completion. The organic solvent was removed by concentration under reduced pressure. Water and ethyl acetate were added and extracted. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a colorless oil, LTL-03-136 (10.47 g, yield 75.75%). LC-MS (ESI) C7H5Br2Cl + [M+H] + Calculated value: 284.84; Measured value: 284.90.

[0453] Step 3: At 0°C, tetrabutylammonium fluoride (1.0 M THF solution, 1.5 eq) was added dropwise to a dichloromethane solution of LTL-03-136 (1.0 eq) and trimethylsilyl cyanide (1.5 eq). The mixture was stirred at 20°C for 1.5 h. The reaction was monitored by LC-MS to indicate completion. The reaction mixture was washed with water, the organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a colorless oil, LTL-03-139 (6.9 g, 81% yield). LC-MS (ESI) C8H5BrClN + [M+H] + Calculated value: 231.93; Measured value: 231.99.

[0454] Step 4: At 0°C, LTL-03-139 (1.00 eq) was dissolved in acetonitrile, and cesium carbonate (3.0 eq) and tert-butyl acrylate (1.1 eq) were added. The mixture was then stirred at 20°C for 6 h. The reaction was monitored by LC-MS until completion. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by pre-silica gel plate analysis to obtain the target product LTL-03-145 (7.59 g, 70.5% yield). LC-MS (ESI) C 15 H 17 BrClNO2 + [M+H] +Calculated value: 360.01; Measured value: 360.33. ¹H NMR (600 MHz, Chloroform-d) δ 7.65 (dd, J = 8.0, 1.5 Hz, 1H), 7.52 (dd, J = 7.8, 1.5 Hz, 1H), 7.22 (t, J = 7.9 Hz, 1H), 4.49 (dd, J = 8.9, 5.7 Hz, 1H), 2.52 – 2.40 (m, 2H), 2.26 –2.11 (m, 2H), 1.45 (s, 9H).

[0455] Step 5: Dissolve LTL-03-145 (1.0 eq) and pinacol diborate (1.5 eq) in 1,4-dioxane, and add potassium acetate (3 eq). Purge three times with nitrogen, add Pd(dppf)Cl2 (0.05 eq), and purge three more times with nitrogen. Heat the reaction mixture to 100°C and stir for 12 h. Detect the completion of the reaction using LCMS, and cool the reaction mixture to room temperature. Add water to the reaction mixture, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain a white solid, LTL-03-172. LCMS (ESI) C 13 H 14 BrN3O + [M+H] + Calculated value: 406.19; Measured value: 406.33.

[0456] Step 6: Dissolve LTL-03-172 (1.0 eq) and LTL-03-065 (1.0 eq) in 1,4-dioxane, and add 2M potassium phosphate solution (2.0 eq). Purge three times with nitrogen, add Pd(dppf)Cl2 (0.05 eq), and purge three more times with nitrogen. Heat the reaction mixture to 90°C and stir for 4 h. Detect the reaction completion by LC-MS. Cool the reaction mixture to room temperature. Add water and ethyl acetate to the reaction mixture for extraction, repeating the extraction three times until complete. Wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue using a pre-prepared silica gel plate to obtain a white solid LTL-03-178 (255 mg, yield 80.96%). LC-MS (ESI) C 32 H 33 ClN4O2 + [M+H] + Calculated value: 541.09; Measured value: 541.24.

[0457] Step 7: Add concentrated sulfuric acid (4.0 eq) to the acetic acid solution of LTL-03-178 (1.00 eq). Stir the mixture at 90 °C for 3 h. LCMS analysis indicates the reaction is complete. Cool to room temperature, pour the reaction mixture into ice water, adjust the pH to 7-8 by adding saturated sodium bicarbonate solution, and extract with dichloromethane. Repeat the extraction three times until complete. Wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain the target product LTL-03-179 as a white solid. LCMS (ESI) C 28 H 25 ClN4O2 + [M+H] + Calculated value: 485.17; Measured value: 485.23. ¹H NMR (600 MHz, DMSO-d6) δ 10.94 (s, 1H), 8.30 (s, 1H), 7.94 (dd, J = 7.3, 2.0 Hz, 1H), 7.90 (d, J = 8.1 Hz, 1H), 7.54 (dd, J = 7.4, 5.5 Hz, 3H), 7.51 – 7.47 (m, 1H), 7.44 – 7.38 (m, 3H), 7.05 (s, 1H), 4.32 (dd, J = 12.4, 5.0 Hz, 1H), 4.16 (s, 2H), 3.77 (s, 2H), 3.71 (t, J = 5.5 Hz, 2H), 2.95(t, J = 5.5 Hz, 2H), 2.78 (ddd, J = 17.8, 13.1, 5.4 Hz, 1H), 2.56 – 2.52 (m,1H), 2.32 (qd, J = 13.0, 4.4 Hz, 1H), 2.04 – 1.99 (m, 1H).

[0458] Example 90: Synthesis of 3-(2-chloro-3-{7-[(2-phenylphenyl)methyl]-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl}phenyl)hexahydropyridine-2,6-dione (LTL-03-184)

[0459] Referring to step 1 of Example 89 and steps 6 and 7 of Example 7, the target product was synthesized by replacing 1-(bromomethyl)naphthalene with 2-(bromomethyl)-1,1'-biphenyl. LCMS (ESI) C 30 H 27 ClN4O2 + [M+H] +Calculated value: 511.18; Measured value: 511.26. ¹H NMR (600 MHz, DMSO-d⁶) δ 9.71 (s, 1H), 6.38 (dd, J = 7.6, 1.6 Hz, 1H), 6.21 (s, 1H), 6.20 – 6.19 (m, 1H), 6.18 – 6.15 (m, 5H), 6.15 – 6.14 (m, 1H), 6.12 (ddd, J = 7.0, 3.3, 2.0 Hz, 2H), 6.03 (dd, J = 7.5, 1.6 Hz, 1H), 5.67 (s, 1H), 3.09 (dd, J = 12.3, 5.1 Hz, 1H), 2.42 (d, J = 7.6Hz, 4H), 2.33 (d, J = 4.6 Hz, 2H), 1.60 – 1.52 (m, 1H), 1.49 (t, J = 5.5 Hz, 2H), 1.31 (dt, J = 17.4, 4.0 Hz, 1H), 1.13 – 1.04 (m, 1H), 0.83 – 0.75 (m, 1H).

[0460] Example 91: Synthesis of 3-{2-chloro-3-[5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl]phenyl}hexahydropyridine-2,6-dione (LTL-04-004)

[0461] Referring to step 1 of Example 89 and steps 6 and 7 of Example 7, the target product was synthesized by replacing 3-bromo-7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine with 3-bromo-5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine. LCMS (ESI) C 28 H 25 ClN4O2 + [M+H] +Calculated value: 485.17; Measured value: 485.36. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.92 (s, 1H), 8.29 – 8.25 (m, 1H), 7.92 – 7.88 (m, 1H), 7.85 (d, J = 8.1 Hz, 1H), 7.60 (s, 1H), 7.50 (ddd, J = 8.8, 6.5, 1.8 Hz, 3H), 7.45 (dd, J = 8.1, 6.9 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H), 7.25 (ddd, J = 16.3, 7.6, 1.8 Hz, 2H), 4.29 (dd, J = 12.3, 5.1 Hz, 1H), 4.12 (s, 2H), 4.10 (t, J = 5.6 Hz, 2H), 3.77 – 3.69 (m, 2H), 2.99 (t, J = 5.6Hz, 2H), 2.80 – 2.72 (m, 1H), 2.57 – 2.52 (m, 1H), 2.27 (qd, J = 12.7, 4.2Hz, 1H), 1.99 (tdd, J = 8.8, 5.3, 3.3 Hz, 1H).

[0462] Example 92: Synthesis of 3-(2-chloro-3-{5-[(2-phenylphenyl)methyl]-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl}phenyl)hexahydropyridine-2,6-dione (LTL-04-005)

[0463] Referring to the method of step 1 in Example 89 and the methods of steps 6 and 7 in Example 7, the target product was synthesized by replacing 3-bromo-7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine with 5-([1,1'-biphenyl]-2-ylmethyl)-3-bromo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine. LCMS (ESI) C 30 H 27 ClN4O2 + [M+H] +Calculated value: 511.18; Measured value: 511.31. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.93 (s, 1H), 7.59 –7.53 (m, 2H), 7.42 (t, J = 7.3 Hz, 2H), 7.37 – 7.30 (m, 6H), 7.27 (dd, J = 7.7, 1.7 Hz, 1H), 7.22 (dd, J = 7.3, 1.7 Hz, 1H), 7.18 (dd, J = 7.5, 1.8 Hz, 1H), 4.30 (dd, J = 12.3, 5.0 Hz, 1H), 4.09 (t, J = 5.7 Hz, 2H), 3.64 (s, 2H), 3.51 – 3.44 (m, 2H), 2.86 – 2.75 (m, 3H), 2.56 – 2.52 (m, 1H), 2.29 (qd, J =12.8, 4.3 Hz, 1H), 2.05 – 1.98 (m, 1H).

[0464] Example 93: Synthesis of 3-(2-chloro-4-(5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)phenyl)piperidine-2,6-dione (LTL-04-109)

[0465] The target product was synthesized according to steps 6-7 of Example 89. LCMS (ESI) C 28 H 25 ClN4O2 + [M+H] +Calculated value: 485.17; Measured value: 485.35. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.91 (s, 1H), 8.33 – 8.28 (m, 1H), 7.93 – 7.90 (m, 1H), 7.89 – 7.84 (m, 2H), 7.55 – 7.46 (m, 4H), 7.44 (d, J = 1.7 Hz, 1H), 7.35 – 7.29 (m, 2H), 4.20 (s, 3H), 4.07 (t, J = 5.6 Hz, 2H), 4.01 (s, 2H), 3.01 – 2.94 (m, 2H), 2.78 (ddd, J = 1.7 Hz, 2H). 17.1,13.0, 5.4 Hz, 1H), 2.54 (dt, J = 17.1, 3.7 Hz, 1H), 2.31 (qd, J = 12.9, 4.3Hz, 1H), 1.99 (dtd, J = 13.2, 5.3, 3.0 Hz, 1H).

[0466] Example 94: Synthesis of 3-(4-(5-([1,1'-biphenyl]-2-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-2-chlorophenyl)piperidine-2,6-dione (LTL-04-110)

[0467] The target product was synthesized according to steps 6-7 of Example 89. LCMS (ESI) C 30 H 27 ClN4O2 + [M+H] +Calculated value: 511.18; Measured value: 511.31. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.91 (s, 1H), 7.83 (s, 1H), 7.59 (dd, J = 7.7, 1.6 Hz, 1H), 7.45 (td, J = 7.3, 1.5 Hz, 2H), 7.40 – 7.35 (m, 5H), 7.35 – 7.32 (m, 2H), 7.24 (td, J = 7.7, 1.8 Hz, 2H), 4.19 (dd, J = 12.5, 5.1 Hz, 1H), 4.07 (t, J = 5.5 Hz, 2H), 3.72 (d, J = 7.7Hz, 4H), 2.85 – 2.74 (m, 3H), 2.61 (t, J = 1.9 Hz, 1H), 2.35 – 2.27 (m, 1H), 2.03 – 1.96 (m, 1H).

[0468] Example 95: Synthesis of 3-(2-chloro-4-(7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)phenyl)piperidine-2,6-dione (LTL-04-097)

[0469] The target product was synthesized according to steps 6-7 of Example 89. LCMS (ESI) C 28 H 25 ClN4O2 + [M+H] +Calculated value: 485.17; Measured value: 485.25. ¹H NMR (600 MHz, DMSO-d⁶) δ 10.92 (s, 1H), 8.35 – 8.31 (m, 1H), 7.94 (dd, J = 7.4, 2.0 Hz, 1H), 7.90 (d, J = 8.2 Hz, 1H), 7.58 (d, J = 1.8 Hz, 1H), 7.56 – 7.53 (m, 3H), 7.51 – 7.48 (m, 1H), 7.45 (dd, J = 8.1, 1.9 Hz, 1H), 7.37 (d, J = 8.1 Hz, 1H), 7.12 (s, 1H), 4.22 (dd, J = 10.92 (s, 1H), 8.35 – 8.31 (m, 1H), 7.94 (dd, J = 7.4, 2.0 Hz, 1H), 7.90 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 8.1 Hz, 1H), 7.12 (s, 1H), 4.22 (dd, J = 10.92 (s, 1H), 8.35 – 8.31 (m ... 12.5, 5.1 Hz, 1H), 4.15 (s, 2H), 4.05 (t, J = 5.5 Hz, 2H), 3.73 (s, 2H), 2.94 – 2.90 (m, 2H), 2.78 (ddd, J = 17.8, 13.1, 5.4 Hz, 1H), 2.55 (d, J = 4.0Hz, 1H), 2.31 (qd, J = 14.6, 13.8, 5.2 Hz, 1H), 2.01 – 1.96 (m, 1H).

[0470] Example 96: Synthesis of 3-(4-(7-([1,1'-biphenyl]-2-ylmethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-2-chlorophenyl)piperidine-2,6-dione (LTL-04-098)

[0471] The target product was synthesized according to steps 6-7 of Example 89. LCMS (ESI) C 30 H 27 ClN4O2 + [M+H] + Calculated value: 511.18; Measured value: 511.27.

[0472] Example 97: Synthesis of (S)-3-(6-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-3-oxo-1,3-dihydro-2H-indazol-2-yl)piperidine-2,6-dione (LC-01-300)

[0473]

[0474] Step 1: Methyl 4-bromo-2-nitrobenzoate (1 eq) and pinacol diboronate (1.5 eq) were dissolved in 1,4-dioxane, and potassium acetate (3 eq) was added. The mixture was purged with nitrogen three times, and Pd(dppf)Cl2 (0.03 eq) was added. The mixture was then purged with nitrogen three times again. The reaction mixture was heated to 100 °C and stirred for 12 h. The reaction was monitored by LC-MS to confirm completion, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain a white solid, LC-01-294 (5.50 g, yield 93.14%). LC-MS (ESI) C 14 H 19 BNO6 + [M+H] + Calculated value: 308.12; Measured value: 308.10.

[0475] Step 2: LC-01-294 (1 eq) and 3-bromo-5,6-dihydroimidazolo[1,2-a]pyrazine-7(8H)-carboxylic acid tert-butyl ester (1 eq) were dissolved in 1,4-dioxane, and 2 M sodium carbonate solution (2 eq) was added. The mixture was purged with nitrogen three times, and Pd(PPh3)4 (0.04 eq) was added. The mixture was then purged with nitrogen three more times. The reaction mixture was heated to 80 °C and stirred for 12 h. The reaction was monitored by LC-MS until complete, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid LC-01-295 (5.60 g, yield 85.48%). LC-MS (ESI) C 19 H 23 N4O6 + [M+H] + Calculated value: 403.15; Measured value: 403.20.

[0476] Step 3: Add LC-01-295 (1 eq) to tetrahydrofuran (30 mL), then add an aqueous solution of lithium hydroxide monohydrate (1.5 eq) (30 mL). Stir overnight at room temperature. Concentrate the reaction solution under reduced pressure and dry to obtain LC-01-296 (4.80 g, yield 99.47%), which was used directly in the next reaction without purification. LC-MS (ESI) C 18 H 21 N4O6 + [M+H] + Calculated value: 389.14; Measured value: 389.20.

[0477] Step 4: LC-01-296 (1 eq), (S)-4,5-diamino-5-oxovalerate tert-butyl hydrochloride (1 eq), and HATU (1 eq) were added to N,N-dimethylformamide (60 mL), followed by dropwise addition of N,N-diisopropylethylamine (3 eq). The reaction mixture was allowed to react at room temperature for 12 h. The reaction solution was diluted with ethyl acetate, washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The remaining residue was subjected to column chromatography to obtain LC-01-297 (6.00 g, yield 84.78%) as a white solid. LC-MS (ESI) C 27 H 37 N6O8 + [M+H] + Calculated value: 573.26; Measured value: 573.30.

[0478] Step 5: Add LC-01-297 (1 eq) to methanol (30 mL), add tetrahydroxydiboron (5 eq) at 0 °C, then add a methanol solution of sodium hydroxide (10 eq) (30 mL), react at 0 °C for 10 minutes, then raise the temperature to 40 °C and react overnight. After the reaction is complete as monitored by TLC, quench the reaction with saturated ammonium chloride solution, extract with ethyl acetate, separate the layers, wash the organic phase with water and saturated sodium chloride solution, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and column chromatography the remaining residue to obtain LC-01-298 (3.50 g, yield 61.79%) as a white solid. LC-MS (ESI) C 27 H 37 N6O6 + [M+H] + Calculated value: 541.27; Measured value: 541.30.

[0479] Step 6: Dissolve LC-01-298 (1 eq) in acetonitrile, add benzenesulfonic acid (3 eq), heat the reaction solution to 100℃, and stir for 12 h. Detect the completion of the reaction using LC-MS. Cool the reaction solution to room temperature, concentrate under reduced pressure to remove the solvent, add ethyl acetate and petroleum ether to the residue, and slurry. Filter to obtain a white solid LC-01-299 (2.00 g, yield 84.32%). LC-MS (ESI) C 18 H 18 N6O3 + [M+H] + Calculated value: 367.14; Measured value: 367.20.

[0480] Step 7: Dissolve LC-01-299 (1 eq) and 2'-chloro-[1,1'-biphenyl]-2-carboxaldehyde (1.2 eq) in N,N-dimethylformimide, and add a catalytic amount of acetic acid. Stir the reaction mixture at room temperature for 30 minutes, then add sodium borohydride acetate (3 eq), and continue stirring for 12 h. Detect the completion of the reaction using LCMS, and quench the reaction mixture with sodium bicarbonate solution. Extract the reaction mixture with ethyl acetate, repeating the extraction three times until complete. Wash the organic phase with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate, and purify using silica gel filtration to obtain a white solid LC-01-300 (32.00 mg, yield 41.35%). LCMS (ESI) C 31 H 28 ClN6O3 + [M+H] + Calculated value: 567.18; Measured value: 567.20.

[0481] The aromatic aldehyde derivatives and aromatic carboxylic acid derivatives used in the embodiments of this invention are synthesized using the following method:

[0482] Option 1: Synthesis of intermediate aromatic aldehyde derivatives (structure shown in Table 1)

[0483] Synthetic Method A: The halogenated substrate (1 eq) and aromatic boric acid (1.2 eq) were dissolved in ethanol, and Na₂CO₃ (1 eq) and Pd(AcO)₂ (0.1 eq) were added to the solution. The reaction system was stirred at room temperature for 12 h under nitrogen protection. The reaction was monitored by thin-layer chromatography. The solid was removed by filtration with diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the intermediate biphenylaldehyde derivative.

[0484] Synthetic Method B: The halogenated substrate (1 eq) and aromatic boric acid (1.05 eq) were dissolved in a mixed solvent of ethanol or methanol, toluene, and aqueous Na₂CO₃. Pd(PPh₃)₄ (0.05 eq) was added to the solvent. Under nitrogen protection, the reaction mixture was heated to 90 °C and stirred for 6 h. The reaction was detected by thin-layer chromatography, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate biphenylaldehyde derivative.

[0485] Synthetic Method C: 2-Bromo-3-pyridinecarboxaldehyde (1 eq) was dissolved in DMF solvent, and K2CO3 (2.5 eq) and 3-methyl-1H-1,2,4-triazole (1 eq) were added to the solvent. The reaction solution was heated to 100 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate aromatic aldehyde derivative.

[0486] Synthetic Method D: 2-Bromobenzaldehyde (1 eq) and piperidin-2-one (1 eq) were dissolved in a dioxane solvent. K3PO4 (2.5 eq), Pd2(dba)3 (0.05 eq), and 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (0.1 eq) were added to the solvent. Nitrogen gas was purged for 5 minutes, and the reaction mixture was heated to 100 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate aromatic aldehyde derivative.

[0487] Synthetic Method E: 2-Bromo-3-pyridinecarboxaldehyde (1 eq) and piperidin-2-one (1 eq) were dissolved in toluene. K3PO4 (2.5 eq), CuI (0.05 eq), and N1,N2-dimethylethane-1,2-diamine (0.1 eq) were added to the solvent. Nitrogen gas was purged for 5 minutes, and the reaction mixture was heated to 100 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the reaction mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate aromatic aldehyde derivative.

[0488] Synthetic Method F: The halogenated substrate (1 eq) and aromatic boric acid (1.2 eq) were dissolved in a mixed solvent of 1,4-dioxane and water. KF (3 eq) and Pd(PPh3)4 (0.05 eq) were added to the solvent. Under nitrogen protection, the reaction solution was heated to 100 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate biphenylaldehyde derivative.

[0489] Synthetic Method G: The halogenated substrate (1 eq) and aromatic boric acid (1.5 eq) were dissolved in N,N-dimethylformimide solution. NaHCO3 (2.00 eq), PPh3 (0.10 eq), and Pd(AcO)2 (0.05 eq) were added to the solvent. Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred for 14 h. The reaction was detected by thin-layer chromatography, and the mixture was cooled to room temperature. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate biphenylaldehyde derivative.

[0490] Synthetic Method H: The halogenated substrate (1 eq) and aromatic boric acid (1.2 eq) were dissolved in a mixed solvent of dimethyl ethylene glycol and water. K₂CO₃ (2.00 eq) and Pd(PPh₃)₄ (0.05 eq) were added to the solvent. Under nitrogen protection, the reaction solution was heated to 85 °C and stirred for 3 h. The reaction was detected by thin-layer chromatography, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the intermediate biphenylaldehyde derivative.

[0491] Table 1: Aromatic aldehyde derivatives synthesized using Scheme 1 in the embodiments of the present invention

[0492]

[0493]

[0494]

[0495] Option 2: Synthesis of intermediate 2'-chloro-[1,1'-biphenyl]-2-carboxylic acid

[0496]

[0497] Step 1: The halogenated substrate (1 eq) and aromatic boric acid (1.05 eq) were dissolved in N,N-dimethylformamide. Pd(AcO)3 (0.05 eq), triphenylphosphine (0.1 eq), and sodium bicarbonate were added to the solvent. Under nitrogen protection, the reaction solution was heated to 110 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain methyl 3-(2-chlorophenyl)pyridinecarboxylate.

[0498] Step 2: Biphenyl carboxylate (1 eq) was dissolved in a mixed solvent of tetrahydrofuran, methanol, and water. Lithium hydroxide (1 eq) was added to the solution at low temperature. The reaction system was stirred at room temperature for 3 h. The reaction was monitored by LCMS to confirm completion. The organic solvent was removed by vacuum concentration. Citric acid was added to the remaining solution to adjust the pH to acidity, resulting in the precipitation of a white solid. The solid was filtered, and the filter cake was concentrated under vacuum to obtain 3-(2-chlorophenyl)pyridinecarboxylic acid.

[0499] Option 3: Synthesis of intermediate 3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde

[0500]

[0501] Step 1: N,N-dimethylformimide (3.00 eq) was dissolved in dichloromethane, and phosphorus tribromide (2.70 eq) was added to the solution at 0 °C. The reaction system was stirred at low temperature for 1 h. Cyclohexanone was added dropwise to the reaction system, and stirring was continued at room temperature for 8 h. The reaction was detected by LCMS to indicate completion. Saturated sodium bicarbonate solution was added to the reaction system for acid-base neutralization, followed by dichloromethane extraction. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2-bromocyclohexyl-1-ene-1-carboxaldehyde.

[0502] Step 2: Dissolve 2-bromocyclohexyl-1-ene-1-carboxaldehyde (1.00 eq) and phenylboronic acid (1.50 eq) in a mixed solvent of 1,4-dioxane and water. Add K₂CO₃ (2.50 eq) and Pd(PPh₃)₄ (0.05 eq) to the solvent. Under nitrogen protection, heat the reaction solution to 100 °C and stir for 12 h. Detect the completion of the reaction by thin-layer chromatography and cool the reaction solution to room temperature. Add water to the reaction solution, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain 3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde.

[0503] Option 4: Synthesis of intermediate 2-(1-bromoethyl)-2'-chloro-1,1'-biphenyl

[0504]

[0505] Step 1: Dissolve 1-(2-bromophenyl)ethyl-1-one (1 eq) and 2-chlorophenylboronic acid (1.05 eq) in a mixed solvent of methanol, toluene, and Na₂CO₃ aqueous solution. Add Pd(PPh₃)₄ (0.05 eq) to the solvent. Under nitrogen protection, heat the reaction solution to 90 °C and stir for 6 h. Detect the completion of the reaction by thin-layer chromatography. Cool the reaction solution to room temperature. Add water to the reaction solution, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain 1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethyl-1-one.

[0506] Step 2: Dissolve 1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethanol-1-one (1.00 eq) in methanol, and add sodium borohydride (2.00 eq) to the solution at 0 °C. Stir the reaction system at low temperature for 6 h. The reaction was confirmed by LCMS. Water was added to the reaction system, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethanol-1-ol. This crude product was directly added to the next step without purification.

[0507] Step 3: Dissolve 1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethanol-1-ol (1.00 eq) in dichloromethane, and add dibromo sulfoxide (3.00 eq) to the solution at 0 °C. Stir the reaction system at room temperature for 8 h. The reaction is complete as detected by LCMS. Add water to the reaction system, extract with ethyl acetate, wash the organic phase with saturated brine, combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography to obtain 2-(1-bromoethyl)-2'-chloro-1,1'-biphenyl.

[0508] Option 5: Synthesis of intermediate 2-(1-methyl-2-oxo-1,2-dihydropyridin-3-yl)benzaldehyde

[0509]

[0510] Step 1: Dissolve 1.00 eq of 3-bromopyridin-2(1H)-one in N,N-dimethylformimide, and add 3.00 eq of CH3I and K2CO3 to the reaction system. Stir the reaction system at 80 °C for 2 h. The reaction was confirmed by LCMS. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, and the two organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 3-bromo-1-methylpyridin-2(1H)-one.

[0511] Step 2: 3-Bromo-1-methylpyridin-2(1H)-one (1.00 eq) and phenylboronic acid (1.20 eq) were dissolved in a mixed solvent of dimethyl ethylene glycol and Na₂CO₃ aqueous solution. Pd(PPh₃)₄ (0.05 eq) was added to the solvent. Under nitrogen protection, the reaction solution was heated to 100 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2-(1-methyl-2-oxo-1,2-dihydropyridin-3-yl)benzaldehyde.

[0512] Option 6: Synthesis of intermediate 4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde

[0513]

[0514] Step 1: N,N-dimethylformimide (3.00 eq) was dissolved in dichloromethane, and phosphorus tribromide (2.70 eq) was added to the solution at 0 °C. The reaction system was stirred at low temperature for 1 h. 4,4-Dimethylcyclohexanone was added dropwise to the reaction system, and stirring was continued at room temperature for 8 h. The reaction was detected by LCMS to indicate completion. Saturated sodium bicarbonate solution was added to the reaction system for acid-base neutralization, followed by dichloromethane extraction. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2-bromo-5,5-dimethylcyclohex-1-ene-1-carboxaldehyde.

[0515] Step 2: 2-Bromo-5,5-dimethylcyclohexyl-1-en-1-carboxaldehyde (1.00 eq) and 4-chlorophenylboronic acid (1.50 eq) were dissolved in a mixed solvent of 1,4-dioxane and water. K₂CO₃ (2.50 eq) and Pd(PPh₃)₄ (0.05 eq) were added to the solvent. Under nitrogen protection, the reaction solution was heated to 100 °C and stirred for 12 h. The reaction was detected by thin-layer chromatography, and the reaction solution was cooled to room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-carboxaldehyde.

[0516] Option 7: Synthesis of intermediate 5-(2-chlorophenyl)pyrimidine-4-carboxaldehyde

[0517]

[0518] Step 1: 5-Bromo-4-methylpyrimidine (1 eq) and (2-chlorophenyl)boronic acid (1.2 eq) were dissolved in a mixed solution of toluene and methanol, and 2 M sodium carbonate solution (2 eq) was added. The mixture was purged three times with nitrogen, and Pd(PPh3)4 (0.04 eq) was added. The mixture was then purged three more times with nitrogen. The reaction mixture was heated to 85 °C and stirred for 12 h. The reaction was monitored by LC-MS until complete, and the mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, and the two phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 5-(2-chlorophenyl)-4-methylpyrimidine (500 mg, yield 42.37%). LC-MS (ESI) C 11 H7ClN2O + [M+H] + Calculated value: 204.66; Measured value: 205.05.

[0519] Step 2: 5-Bromo-4-methylpyrimidine (1 eq) was dissolved in 1,4-dioxane, and selenium dioxide (5 eq) was added. The reaction mixture was heated to 110 °C and stirred for 12 h. The reaction was monitored by LC-MS until completion, and the reaction mixture was cooled to room temperature. Water and ethyl acetate were added to the reaction mixture for extraction, and the extraction was repeated three times until complete. The organic phase was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give 5-(2-chlorophenyl)pyrimidine-4-carboxaldehyde (320 mg, yield 59.93%). LC-MS (ESI) C 11 H9ClN2 + [M+H] + Calculated value: 218.64; Measured value: 219.03.

[0520] Example 98: Construction and testing of the NEK7-N-HiBiT cell line

[0521] 1. Construction of the NEK7-N-HiBiT cell line

[0522] Combine 2 μg NLS-Cas9-EGFP nuclease (Novoprotein, #E379-01A), 50 pmole sgRNA (5'-mG*mU*mU*CCUGU UGCUU CAGAC AAGUU UUAGA GCUAG AAAUA GCAAG UUAAA AUAAG GCUAGUCCGU UAUCA ACUUG AAAAA GUGGC ACCGA GUCGG UGCU*mU*mU*mU-3' (SEQ ID NO: 1)) and 50pmole Donor DNA (5'- TTACA TTTTT GCAGA GTTCT AAAGT TCCTG TTGCT TCAGA CAATGGTCTC CGTGA GCGGC TGGCG GCTGT TCAAG AAGAT TAGCG ATGAG CAATC ACAAG GAATG CAAGGGCCAC CTGTT CCTC-3' (SEQ ID NO: 2)) Add 10μL electroporation buffer. After 10 minutes of incubation, 2 million MOLT4 cells were resuspended in 10 μL of electroporation buffer and then mixed with 10 μL of ribonucleoprotein (RNP) complex. The RNP complex was then electroporated into the cells using a Celestix SP100 electroporator. The cells were then transferred to antibiotic-free 1640 complete medium and incubated overnight. EGFP-positive cells were sorted using a Sony MA900 cell sorter, and positive single-cell clones were selected using a Nano-Glo HiBiT lysis detection kit (Promega, #N3030).

[0523] 2. Compound testing

[0524] Dilute positive cells to 1×10⁻⁶ 5 The concentration of the compound was 100 μL / well in a 96-well plate, followed by the addition of diluted compound. The control group received 0.1% DMSO. The plates were incubated for 6 h. The lysis was detected using a Nano-Glo HiBiT lysis detection kit, and data were analyzed using Graphpad Pism 9.5. Results are shown in Table 2. Figure 1 .

[0525] Table 2: Degradation bioactivity of the compounds of the present invention in Molt4-NEK7-Hibit cells

[0526]

[0527]

[0528]

[0529]

[0530] In Table 2, A represents D. max ≤30%; B represents 30%. <D max ≤70%; C represents D max >70%; + indicates DC 50 >500nM;++ indicates 200nM < DC 50 ≤500nM; +++ indicates DC 50 ≤200nM; - indicates not detected.

[0531] 3. Phenotypic verification of NEK7 degrading agent

[0532] 3.1 Construction of Inflammation Model

[0533] Human peripheral blood mononuclear cells (PBMCs) were isolated from fresh human whole blood, and further isolated into monocytes. The cells were resuspended in RPMI-1640 complete medium and the density was adjusted to 1×10⁶ cells / mL. 6 Macrophages were seeded at a density of 1 / mL into 10 cm culture dishes and M-CSF was added. Induction was performed for 6 days. On the sixth day, macrophages were collected by trypsin digestion and seeded into 96-well plates. After overnight cell adhesion, the compound and MCC950 were added to the wells, while 0.1% DMSO in 1640 medium was added to the control wells. The plates were incubated for 12 h. 500 ng / mL LPS was added for 3 h, followed by Nigericin (final concentration 5 μM) for 1 h. The supernatant was collected for the determination of IL-1β and IL-18 secretion levels.

[0534] 3.2 ELISA detection of IL-1β secretion

[0535] The secretion of IL-1β was detected using the Human IL-1β ELISA Kit (EK101B) from Linko Biotechnology.

[0536] (1) Experimental preparation

[0537] Before testing, please bring all reagents and samples to room temperature (25℃±3℃). If concentrated reagents crystallize, incubate at 37℃ until all crystals dissolve.

[0538] 1× Washing Solution: Pipette 50 mL of 20× concentrated washing solution into a 1 L graduated cylinder, add distilled water to 1,000 mL, and mix gently, avoiding foaming. Transfer to a clean bottle and store at 2-8℃. 1× washing solution is stable for 30 days.

[0539] 1× Detection Buffer: Pipette 5 mL of 10× concentrated detection buffer into a 100 mL graduated cylinder, add distilled water to 50 mL, mix gently to avoid foaming, and store at 2-8℃. 1× detection buffer can be stably stored for 30 days.

[0540] Antibody for testing: Mix thoroughly before dilution. Dilute the concentrated antibody 1:100 with 1× test buffer, depending on the number of standards and samples to be tested (use within 30 minutes).

[0541] Horseradish peroxidase-labeled streptavidin: Mix thoroughly before dilution. Dilute the concentrated horseradish peroxidase-labeled streptavidin 1:100 with 1× detection buffer according to the number of standards and test samples (use within 30 min).

[0542] Standards: Briefly centrifuge before opening, then reconstitute the standard with distilled water, labeling the reconstituted volume on the standard label. Gently vortex to ensure thorough mixing; the reconstituted standard concentration is 500 pg / mL. Let stand for 10-30 minutes after reconstitution. Mix thoroughly before dilution. Take 200 μL of concentrated human IL-1β standard and add 200 μL of standard diluent to obtain the highest concentration (250 pg / mL) of the standard curve. Add 200 μL of standard diluent to each test tube. Perform 1:1 serial dilutions using the high-concentration standard. Ensure thorough mixing after each pipetting. Use the standard diluent as the zero concentration of the standard curve. Take 200 μL of concentrated human IL-1β standard and add 200 μL of cell culture medium to obtain the highest concentration (IL-1β: 250 pg / mL) of the standard curve. Add 200 μL of cell culture medium to each test tube. Perform 1:1 serial dilutions using the high-concentration standard. Ensure thorough mixing with each pipette. Use cell culture medium as the zero concentration for the standard curve.

[0543] (2) Formal testing

[0544] Before testing, equilibrate all reagents and samples to room temperature (25℃±3℃).

[0545] 1) Prepare all necessary reagents and working concentration standards.

[0546] 2) Remove the unwanted slats, put them back into the aluminum foil bag containing the desiccant, and reseal the bag.

[0547] 3) Add 300 μL of 1× washing buffer and let it stand for 30 seconds. Soaking is necessary to obtain ideal experimental results. After discarding the washing buffer, pat the microplate dry on absorbent paper. Use the microplate immediately after washing; do not allow it to dry.

[0548] 4) Add 100 μL of 2-fold serially diluted standard to the standard wells. Add 100 μL of standard dilution buffer (serum / plasma sample) or culture medium (cell culture supernatant sample) to the blank wells.

[0549] 5) Add 100 μL of cell culture supernatant to each sample well.

[0550] 6) Add 50 μL of diluted detection antibody (1:100) to each well. Ensure that steps 4, 5, and 6 are performed consecutively without interruption. The sample addition process should be completed within 15 minutes.

[0551] 7) Seal the plate using a sealing film. Shake at 100-300 rpm (ensure that the solution in each well is not spilled and is thoroughly mixed), and incubate at room temperature (25℃±3℃) for 2 hours.

[0552] 8) Discard the liquid, add 300 μL of washing buffer to each well, and wash the plate 6 times. After each wash, pat the plate dry on absorbent paper. To obtain ideal experimental performance, residual liquid must be completely removed.

[0553] 9) Add 100 μL of diluted horseradish peroxidase-labeled streptavidin (1:100) to each well.

[0554] 10) Seal the plate using a new sealing film. Shake at 100-300 rpm (ensure that the solution in each well is not spilled and is thoroughly mixed), and incubate at room temperature (25℃±3℃) for 45 minutes.

[0555] 11) Repeat step 8).

[0556] 12) Add 100 μL of chromogenic substrate TMB to each well, protect from light, and incubate at room temperature (25℃±3℃) for 5-30 minutes.

[0557] 13) Add 100 μL of stop solution to each well. The color will change from blue to yellow. If the color is green or the color change is significantly uneven, gently tap the plate frame to mix thoroughly.

[0558] 14) Within 30 minutes, perform dual-wavelength detection using a microplate reader, measuring the OD value at the maximum absorption wavelength of 450 nm and the reference wavelength of 570 nm or 630 nm. The calibrated OD value is the 450 nm measurement value minus the 570 nm or 630 nm measurement value. Using only 450 nm for measurement will result in an overestimation of the OD value and reduced accuracy.

[0559] 15) First, fit the standard curve, then generate the sample concentration. Data analysis was performed using Graphpad Pism 9.5; the results are shown in Table 3 and... Figure 2 .

[0560] 3.3 ELISA detection of IL-18 secretion

[0561] The amount of IL-18 secreted was detected using the Human IL-18 ELISA Kit (EK118 / 2) from Linko Biotechnology.

[0562] (1) Experimental preparation

[0563] Before testing, please bring all reagents and samples to room temperature (25℃±3℃). If concentrated reagents crystallize, incubate at 37℃ until all crystals dissolve.

[0564] 1× Washing Solution: Pipette 50 mL of 20× concentrated washing solution into a 1 L graduated cylinder, add distilled water to 1,000 mL, and mix gently, avoiding foaming. Transfer to a clean bottle and store at 2-8°C. 1× washing solution is stable for 30 days.

[0565] 1× Detection Buffer: Pipette 5 mL of 10× concentrated detection buffer into a 100 mL graduated cylinder, add distilled water to 50 mL, mix gently to avoid foaming, and store at 2-8℃. 1× detection buffer can be stably stored for 30 days.

[0566] Antibody for testing: Mix thoroughly before dilution. Dilute the concentrated antibody 1:100 with 1× test buffer according to the number of standards and samples to be tested (use within 30 minutes).

[0567] Standards: Briefly centrifuge before opening, then reconstitute the standard with distilled water, labeling the reconstituted volume on the standard label. Gently vortex to ensure thorough mixing; the reconstituted standard concentration is 20 ng / mL. Let stand for 10-30 minutes after reconstitution. Mix thoroughly before dilution. Take 200 μL of concentrated human IL-18 standard and add 200 μL of standard diluent to obtain the highest concentration of the standard curve (IL-18: 10 ng / mL). Add 200 μL of standard diluent to each test tube. Perform 1:1 serial dilutions using the high-concentration standard. Ensure thorough mixing after each pipetting. Use the standard diluent as the zero concentration of the standard curve. Take 200 μL of concentrated human IL-18 standard and add 200 μL of cell culture medium to obtain the highest concentration of the standard curve (IL-18: 10 ng / mL). Add 200 μL of cell culture medium to each test tube. Perform 1:1 serial dilutions using the high-concentration standard. Ensure thorough mixing with each pipette. Use cell culture medium as the zero concentration for the standard curve.

[0568] (2) Formal testing

[0569] Before testing, equilibrate all reagents and samples to room temperature (25℃±3℃).

[0570] 1) Prepare all necessary reagents and working concentration standards.

[0571] 2) Remove the unwanted slats, put them back into the aluminum foil bag containing the desiccant, and reseal the bag.

[0572] 3) Add 300 μL of 1× washing buffer and let it stand for 30 seconds. Soaking is necessary to obtain ideal experimental results. After discarding the washing buffer, pat the microplate dry on absorbent paper. Use the microplate immediately after washing; do not allow it to dry.

[0573] 4) Add 100 μL of 2-fold serially diluted standard to the standard wells. Add 100 μL of standard dilution buffer (serum / plasma sample) or culture medium (cell culture supernatant sample) to the blank wells.

[0574] 5) Add 100 μL of cell culture supernatant to each sample well.

[0575] 6) Add 50 μl of diluted detection antibody (1:100) to each well. Ensure that steps 4, 5, and 6 are performed consecutively without interruption. The sample addition process should be completed within 15 minutes.

[0576] 7) Seal the plate using a sealing film. Shake at 100-300 rpm (ensure that the solution in each well is not spilled and is thoroughly mixed), and incubate at room temperature (25℃±3℃) for 2 h.

[0577] 8) Discard the liquid, add 300 μL of washing buffer to each well, and wash the plate 6 times. After each wash, pat the plate dry on absorbent paper. To obtain ideal experimental performance, residual liquid must be completely removed.

[0578] 9) Add 100 μL of chromogenic substrate TMB to each well, protect from light, and incubate at room temperature (25℃±3℃) for 5-30 minutes.

[0579] 10) Add 100 μL of stop solution to each well. The color will change from blue to yellow. If the color is green or the color change is significantly uneven, gently tap the plate frame to mix thoroughly.

[0580] 11) Within 30 minutes, perform dual-wavelength detection using a microplate reader, measuring the OD value at the maximum absorption wavelength of 450 nm and the reference wavelength of 570 nm or 630 nm. The calibrated OD value is the 450 nm measurement value minus the 570 nm or 630 nm measurement value. Using only 450 nm for measurement will result in an overestimation of the OD value and reduced accuracy.

[0581] 12) First, fit the standard curve, then generate the sample concentrations. Data analysis was performed using Graphpad Pism 9.5; the results are shown in Table 3 and... Figure 3.

[0582] Table 3: Degradation bioactivity of the compounds of the present invention in Molt4-NEK7-Hibit cells

[0583]

[0584] As shown in Table 3, compound LC-06-014 significantly downregulated the levels of cytokines IL-1β and IL-18 in human primary peripheral blood mononuclear cells (hPBMCs).

[0585] Figures 1-3 The results of HiBiT detection of the compounds of the present invention, as well as the detection results of IL-1β and IL-18, are shown.

[0586] The above results indicate that the compounds of the present invention can inhibit the activation of the NLRP3 inflammasome complex by targeting and degrading NEK7, thereby regulating the secretion of IL-1β and IL-18, and thus can be used to prevent and / or treat diseases related to NEK7 protein function or IL-1β / IL-18-mediated diseases.

[0587] The above detailed embodiments describe the implementation of the present invention; however, the present invention is not limited to the specific details described in the above embodiments. Within the scope of the claims and technical concept of the present invention, various simple modifications and changes can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. An aromatic heterocyclic compound based on glutarimide, characterized in that, It has the structure shown in equation (I): ； In formula (I): R 1a R 1b R 1c R 1d Each is independently selected from hydrogen, deuterium, halogen, and halogenated carbon. 1-10 Alkyl, deuterated C 1-10 Alkyl, C 1-10 Alkoxy, deuterated C 1-10 alkoxy or halogenated C 1-10 Alkoxy; Y is selected from CH, CD, CF, CMe, or N; X is a chemical bond; Cy1 is selected from a monocyclic, bicyclic, or tricyclic skeleton having n substituents R4 or nitrogen atom substitutions. Each substituent R4 may be the same or different and is independently selected from H, deuterium, halogen, -CH3, -CF3, alkynyl, -OH, -SH, -NO2, -NH2, -CN, oxo, or C. 1-6 alkyl; Cy2 is selected from unsubstituted or pentanehexasaturated bicyclic or pentanehexasaturated bicyclic with m substituents; each substituent may be the same or different and is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -NO2, -CF2H, oxo, aromatic benzyl, aryl, heteroaromatic or C 1-10 alkyl; R w Selected from one or more of H, -CO-W, -CH(CH3)-W, -CH2-W, and -SO2-W; W is a substituted or unsubstituted cyclic hydrocarbon group, saturated heterocyclic group, aryl, heteroaryl, naphthyl, biphenyl, etc. , , , , , , Benzenecycloaryl, thiazolyl, or benzoxane; T1, T2, T3, and T4 are each independently selected from CH or N; R x Selected from H, halogen, -CF3, C 1-7 Alkyl and R y C with substituent group 1-7 Alkyl or aryl; R y Selected from H, F, Cl, -OCH3, -CN, -CF2H, -OCF3, -OPh, -NHPh, -OCH2Ph, -CH3, or -CF3.

2. The aromatic heterocyclic compound based on glutarimide according to claim 1, characterized in that, The ring Cy1 is selected from, but is not limited to, single-ring, double-ring, or triple-ring skeletons with the following structural forms: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ; Among them, R 2a R 3a Selected from H or C 1-6 Alkyl; R 2b Selected from F or CH3; R 2c Selected from H or C 1-6 Alkyl; R 4a R 4b Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -OCH3, -OCF3, -OCF2H, alkynyl, or C. 1-6 Alkyl group; the symbol * indicates the junction of the ring Cy1 and X, and the symbol ** indicates the junction of the ring Cy1 and Cy2.

3. The aromatic heterocyclic compound based on glutarimide according to claim 1, characterized in that, The Cy2 ring is selected from, but is not limited to, pentahexasaturated bicyclic or pentahexas unsaturated bicyclic structures: , , , , , , , , , , , or ; Among them, R 5a Selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, aromatic benzyl, aryl, or C. 1-6 Alkyl; R 5b R 5c R 5d R 5e Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -OCF3, -OCH3, -CF2H, -OCF2H, or C. 1-10 Alkyl; Z and R 5f The dashed line between them indicates that there is R. 5f Replacement or no R 5f Replacement, when there is R 5f When replacing, Z is N, R 5f Selected from H, -CH3, -CF3, -CH2CF3, aromatic benzyl, aromatic group, heteroaromatic group, or C. 1-10 Alkyl; when R is absent 5f When replacing, Z is O or S; R 6a R 6b R 6c R 6d R 6e Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl; R 7a R 7b R 7c R 7d Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl; R 8a R 8b R 8c Each is independently selected from H, deuterium, halogen, -CH3, -CF3, -CN, -NH2, -OCF3, -OCH3, -NO2, -CF2H, -OCF2H, or C. 1-6 Alkyl group; the symbol * indicates the junction of the ring Cy2 and the ring Cy1, and the symbol ** indicates the junction of the ring Cy2 and the group R. w The connection point.

4. The aromatic heterocyclic compound based on glutarimide according to claim 1, characterized in that, The halogens include fluorine, chlorine, bromine, or iodine; The alkyl group includes straight-chain alkyl, branched alkyl, or cycloalkyl; The aryl group includes phenyl or fused-ring aryl; The heteroaryl group includes pyrrole, thiophene, furanyl, thiazolyl, pyrazolyl, imidazole, pyridinyl, pyridinyl, pyrazinyl, oxazolyl, or pyrimidinyl. The cyclic hydrocarbon group is a cycloalkane having 3 to 7 carbon atoms; The saturated heterocyclic group is a saturated five- to seven-membered ring group having one or two heteroatoms, and the heteroatoms are one or more of O, N, and S.

5. The aromatic heterocyclic compound based on glutarimide according to claim 4, characterized in that, The fused-ring aryl group includes fused-ring alkyl phenyl, fused-ring heteroaryl, fused-ring heteroaryl phenyl, fused-ring heteroaryl heteroaryl, fused-ring alkyl naphthyl, fused-ring heteroaryl naphthyl, fused-ring alkyl anthracel, or fused-ring heteroaryl anthracel.

6. The aromatic heterocyclic compound based on glutarimide according to claim 1, characterized in that, The aromatic heterocyclic compounds based on glutarimide are selected from the following compounds: (S)-3-(1-oxo-5-(7-((2',3',4',5'-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidin-2,6-dione; (S)-3-(1-oxo-5-(7-((3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazol[1,2-a]pyrazin-3-yl)isoindol-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazol[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2',3'-dichloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2',4'-dichloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-3'-methoxy-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-3'-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-2-chloro-2'-((3-(2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-[1,1'-biphenyl]-3-cyano; (S)-3-(5-(7-((2'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-4'-methoxy-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazolium[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-2-chloro-2'-((3-(2-(2-(2-6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazolium[1,2-a]pyrazin-7(8H)-yl)methyl)-[1,1'-biphenyl]-4-cyano; (S)-3-(5-(7-((2'-chloro-5'-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-5'-methoxy-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-6-chloro-2'-((3-(2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-[1,1'-biphenyl]-3-cyano; (S)-3-(5-(7-((2',5'-dichloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-(2-(6-chlorobenzo[d][1,3]dioxo-5-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-3-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-4-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-5-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-6-fluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-3,4-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-3,6-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-4,5-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-4,6-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-5,6-difluoro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-8-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-(1-(2'-chloro-[1,1'-biphenyl]-2-yl)ethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-3-methyl-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-3-methyl-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-8,8-dimethyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; 5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-((3-(2-chlorophenyl)pyridin-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((4-(2-chlorophenyl)pyridin-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((3-(2-chlorophenyl)pyridin-4-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2-(2-chlorophenyl)pyridin-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((5-(2-chlorophenyl)pyrimidin-4-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-imidazol[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-2-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-2-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-6-fluoro-1-oxoisoindol-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2-(3-methyl-1H-1,2,4-triazol-1-yl)pyridin-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(1-oxo-5-(7-(2-(2-oxopiridine-1-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-(2-(1-methyl-2-oxo-1,2-dihydropyridin-3-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindol-2-yl)piperidin-2,6-dione; (S)-3-(1-oxo-5-(7-((2-oxo-2H-[1,2'-bipyridin]-3'-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-((2'-bromo-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-3-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-(3-(2-chlorophenyl)pyridinyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-(3-(2-chlorophenyl)pyridinyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-4-fluoro-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-(2'-chloro-[1,1'-biphenyl]-2-carbonyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(2-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-5-oxo-5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)piperidine-2,6-dione; (S)-3-(5-(5-(2-(benzyloxy)benzyl)-4,5,6,7-tetrahydro-1H-[1,2,3]triazolo[4,5-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(5-([1,1'-biphenyl]-2-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-1-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(5-([1,1'-biphenyl]-2-ylmethyl)-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindol-2-yl)piperidin-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(5-([1,1'-biphenyl]-2-ylmethyl)-3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(5-(3-benzyl-5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-1-methyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (3S)-3-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4-methyl-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-cyclopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(5-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-3-phenyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(6-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridin-3-yl)-1-oxoisoindololin-2-yl)piperidine-2,6-dione; (S)-3-(5-(5-benzyl-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(1-oxo-5-(7-(2-oxo-1,2-dihydropyridin-3-yl)benzyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)-7-fluoro-1-oxoisoindol-2-yl)piperidin-2,6-dione; (S)-3-(5-(7-((S)-2-(benzylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-3-carbonyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-N-(2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)phenyl)acetamide; (S)-3-(5-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)sulfonyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-N,N-dimethylbenzamide; (S)-2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)methyl)-N-methylbenzamide; (S)-3-(5-(6-benzyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(6-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(6-benzyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; (S)-3-(5-(6-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidine-2,6-dione; (S)-3-(1-oxo-5-(7-(benzenesulfonyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)isoindololin-2-yl)piperidin-2,6-dione; (S)-3-(5-(6-benzyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[2,3-c]pyridin-3-yl)-1-oxoisoindoline-2-yl)piperidin-2,6-dione; 3-{5-[7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl]-1-benzofuran-3-yl}hexahydropyridine-2,6-dione; 3-(5-{7-[(2-phenylphenyl)methyl]-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl}-1-benzofuran-3-yl)hexahydropyridine-2,6-dione; 3-{5-[5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl]-1-benzofuran-3-yl}hexahydropyridine-2,6-dione; 3-(5-{5-[(2-phenylphenyl)methyl]-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl}-1-benzofuran-3-yl)hexahydropyridine-2,6-dione; 3-{6-[7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl]-1-benzofuran-3-yl}hexahydropyridine-2,6-dione; 3-(6-{7-[(2-phenylphenyl)methyl]-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl}-1-benzofuran-3-yl)hexahydropyridine-2,6-dione; 3-{2-chloro-3-[7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl]phenyl}hexahydropyridine-2,6-dione; 3-(2-chloro-3-{7-[(2-phenylphenyl)methyl]-5,6,7,8-tetrahydroimidazo[3,2-a]pyrazin-3-yl}phenyl)hexahydropyridine-2,6-dione; 3-{2-chloro-3-[5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl]phenyl}hexahydropyridine-2,6-dione; 3-(2-chloro-3-{5-[(2-phenylphenyl)methyl]-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl}phenyl)hexahydropyridine-2,6-dione; 3-(2-chloro-4-(5-(naphth-1-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)phenyl)piperidine-2,6-dione; 3-(4-(5-([1,1'-biphenyl]-2-ylmethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-3-yl)-2-chlorophenyl)piperidine-2,6-dione; 3-(2-chloro-4-(7-(naphth-1-ylmethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)phenyl)piperidine-2,6-dione; 3-(4-(7-([1,1'-biphenyl]-2-ylmethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-2-chlorophenyl)piperidine-2,6-dione; (S)-3-(6-(7-((2'-chloro-[1,1'-biphenyl]-2-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)-3-oxo-1,3-dihydro-2H-indazole-2-yl)piperidine-2,6-dione.

7. A pharmaceutical composition, characterized in that, The compound comprises an effective amount of one or more of the aromatic heterocyclic compound based on glutarimide as described in any one of claims 1 to 6, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, deuterated compound, or polymorph thereof.

8. Use of the aromatic heterocyclic compound based on glutarimide according to any one of claims 1 to 6 in the preparation of a medicament for the prevention and / or treatment of diseases related to the NEK7 protein.

9. Use of the aromatic heterocyclic compound based on glutarimide according to any one of claims 1 to 6 in the preparation of a medicament for the prevention and / or treatment of diseases related to the NLRP3 inflammasome.

10. Use of the aromatic heterocyclic compound based on glutarimide according to any one of claims 1 to 6 in the preparation of a medicament for the prevention and / or treatment of IL-1β and / or IL-18-mediated related diseases.

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