A small molecule degrader of arylformamidines and uses thereof

Abstract

The application discloses a small molecule degrading agent of aryl formamides and application thereof, and has a structure shown in general formula I or a pharmaceutically acceptable salt, an isotopically labeled compound thereof. The compound of the application has a targeted degradation activity on MYC protein, and has a significant proliferation inhibiting activity on various solid tumor cells such as neuroblastoma and rhabdomyosarcoma. The compound of the application can be used for preparing a degrading agent targeting MYC protein, and can also be used for preparing a drug for treating diseases related to MYC gene amplification, fusion, mutation, expression imbalance, conformational change and function abnormality.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, specifically to MYC-targeting small molecule degraders, pharmaceutical compositions thereof, and their uses. Background Technology

[0002] MYC proteins, as key oncogenic transcription factors, are classified into N-Myc, C-Myc, and L-Myc. MYC proteins participate extensively in cell proliferation, cell cycle, angiogenesis, and anti-tumor immunity by regulating the expression of downstream target genes. They play a crucial role in the occurrence and development of various malignant tumors, such as neuroblastoma and rhabdomyosarcoma. Therefore, targeting MYC is considered an important strategy for cancer treatment; however, drug development has been slow, and it has long been considered an "untreatable" target. On the one hand, due to its highly disordered nature and lack of a stable three-dimensional conformation, no experimentally resolved structural information has been obtained for MYC proteins, limiting the rational design of targeted small molecules. On the other hand, other indirect intervention methods, such as disrupting the function of the MYC transcription complex, have limitations such as poor selectivity, significant toxicity, and poor efficacy; currently, no effective drugs have been marketed. Furthermore, studies have found that KLHL37 binding to N-Myc prevents the degradation of MYC proteins mediated by E3 ubiquitinase, leading to increased stability and abnormal activation of N-Myc proteins, which is also a major reason for the malignant progression and poor prognosis of tumors such as neuroblastoma. Therefore, there is an urgent need to develop effective small molecules that can directly target the MYC protein.

[0003] MYC protein is mainly degraded in cells through the ubiquitin-proteasome pathway. Therefore, intervening in MYC protein stability has become another potential intervention method, and it is hoped that new intervention molecules can be developed to treat diseases related to MYC gene amplification, fusion, mutation, expression imbalance, allosteric changes and functional abnormalities. Summary of the Invention

[0004] This invention provides a compound with targeted degradation activity against MYC protein and significant anti-proliferative inhibitory activity against various solid tumor cells, including neuroblastoma and rhabdomyosarcoma. The compound of this invention can be used to prepare MYC protein-targeting degradative agents, and also to prepare drugs for treating diseases related to MYC gene amplification, fusion, mutation, expression imbalance, allosteric changes, and functional abnormalities.

[0005] On one hand, the present invention provides a compound having the structure shown in general formula I or a pharmaceutically acceptable salt or isotope-labeled compound thereof.

[0006]

[0007] A represents a substituted or unsubstituted acetamide group, or a cyclized amide or amino group;

[0008] B is an unsubstituted, substituted acetamide subunit, or absent;

[0009] M is a carbon or nitrogen atom;

[0010] C is hydrogen, halogen, heterocyclic alkyl, aryl, heteroaryl, or absent;

[0011] R1 and R2 are each independently hydrogen, alkyl, or R1 and R2 together with the atoms they are attached to form a 5-7 membered ring; the 5-7 membered ring is optionally substituted by one or more substituents selected from alkyl groups or heteroatom-containing alkyl groups.

[0012] D represents a structure that binds to E3 ubiquitin ligase or heat shock protein, or it may not exist.

[0013] For substituent A, when substitution is present, the substituent is hydrogen, C1-C5 alkyl, C1-C5 heterocyclic alkyl, C6-C10 aryl, C1-C3 alkoxy, or halogen; the substituent is further preferably phenyl, piperazine, etc.; the cyclized amide group is preferably 1-isoindololinone-2-yl.

[0014] For substituent B, when substitution is present, the substituent is C1-C5 alkyl, C6-C10 aryl, or C1-C3 alkoxy; the substituent is further preferably methyl, etc.

[0015] For the substituent C, the heterocyclic alkyl group is preferably a 3- to 8-membered heterocyclic alkyl group; the aryl group is preferably a C6- to C10 aryl group; the heterocyclic alkyl group, aryl group, and heteroaryl group may be further substituted by C1- to C3 alkyl groups or C1- to C3 alkoxy groups. The heterocyclic alkyl group is preferably tetrahydropyridyl, and the aryl group is preferably pyridyl or phenyl; wherein the substituent is preferably methyl, methoxy, etc.

[0016] In R1 and R2, the alkyl group is a C1 to C5 alkyl group; the alkyl group may be further replaced by a 3 to 8-membered heterocyclic alkyl group or an amino group; the 5-7-membered ring is preferably a piperazine ring, a piperidine ring, etc.; the substituent is preferably methyl or morpholino.

[0017] In some embodiments,

[0018] A is Wherein, R3 is hydrogen, C1-C5 alkyl, C1-C5 alkylhexyl, C6-C10 aryl; E is C6-C10 aromatic ring, 5-6 membered heteroaromatic ring or absent; R4 is hydrogen, C1-C3 alkyl, C1-C3 alkoxy or halogen; n = 1-2 (n is 1 or 2).

[0019] B is Or not present; wherein, R5 is C1-C5 alkyl, C6-C10 aryl, C1-C3 alkoxy; m = 1-5 (m is 1, 2, 3, 4 or 5); *the corresponding C atom is S configuration or R configuration or a mixture of S configuration and R configuration.

[0020] C is hydrogen, halogen, C4-C6 heterocyclic alkyl, C6-C10 aryl, 5-6 heteroaryl or absent; wherein, the C6-C10 aryl or 5-6 heteroaryl is independently substituted by one or more substituents selected from halogen, hydroxyl, C1-C3 alkyl or C1-C3 alkoxy.

[0021] for Among them, R1 and R6 are each independently hydrogen and C1 to C5 alkyl groups; R7 and R8 are each independently -CH2-, -NH-, or -NR. 11 -, O, S; R9 is H, C1-C5 alkyl, and there can be two or more R9s; Q is -CH- or -N-; x = 1-5; y = 1-4; z = 1-4 (z is 1, 2, 3 or 4); R 11 It is a C1 to C3 alkyl group.

[0022] D represents a small ligand that binds to CRBN; a small ligand that binds to VHL; a covalent fragment that binds to other E3 ubiquitin ligases; and a small ligand that binds to HSP70.

[0023] In some embodiments, the compounds of the present invention have a structure shown in any one of general formulas I-1 to I-16, or a pharmaceutically acceptable salt or isotope-labeled compound thereof:

[0024]

[0025] R1, R3, R5, R6, R9, x, Same as the definition in general formula I; x a x b x c x d x e Each of the following groups is independently selected from hydrogen, halogen, cyclopropyl, cyano, carbonyl, carbamoyl, and C6-C10 arcarbamoyl; wherein each of the C6-C10 arcarbamoyl groups is independently substituted by one or more substituents selected from halogen, hydroxyl, C1-C3 alkyl, or C1-C3 alkoxy.

[0026] Preferably, the compound has the structure shown in the following general formula:

[0027] Where R 22 Selected from H,

[0028] As a preferred choice, A is selected from Or amino.

[0029] As a preferred choice, B is selected from Or it may not exist.

[0030] As a preferred option, C is... H or Br.

[0031] As a preferred option, D is H.

[0032] As a preferred option Selected from -NHMe,

[0033] In some embodiments, the compound has one of the following structures or a pharmaceutically acceptable salt or isotope-labeled compound thereof;

[0034]

[0035]

[0036]

[0037] On the other hand, the present invention provides a compound for preparing a degrader targeting MYC protein, and can also be used to prepare a drug for treating diseases related to MYC gene amplification, fusion, mutation, expression imbalance, allosteric changes and functional abnormalities.

[0038] The compounds of this invention exhibit targeted degradation activity against MYC protein and significant inhibitory activity against the proliferation of various solid tumor cells, including neuroblastoma and rhabdomyosarcoma. These compounds can be used to prepare MYC protein-targeting degradative agents and drugs for treating diseases related to MYC gene amplification, fusion, mutation, expression imbalance, allosteric changes, and functional abnormalities. Attached Figure Description

[0039] Figure 1 The affinity of the compounds provided in this invention for N-Myc and C-Myc proteins was determined by the MST method.

[0040] Figure 2 Changes in N-Myc protein levels in neuroblastoma amplified by MYCN after compound treatment.

[0041] Figure 3Tumor inhibitory effect of compound A12 in the RH30 xenograft tumor model. A: RH30 tumor volume change over time; B: Photographs of tumors in each group at the experimental endpoint; C: Statistical analysis of tumor weight in each group.

[0042] Figure 4 Tumor inhibitory effect of compound A12 in the SK-B-DZ xenograft model. A: SK-N-DZ tumor volume change over time; B: Photographs of tumors in each group at the experimental endpoint; C: Statistical analysis of tumor weight in each group.

[0043] Figure 5 The tumor-suppressive effect of compound A12 in the SK-B-BE(2) xenograft tumor model. A: SK-N-BE(2) tumor volume change over time curve; B: Photographs of tumors in each group at the experimental endpoint; C: Statistical analysis of tumor weight in each group.

[0044] Figure 6 Tumor inhibitory effect of compound A12 in a patient-derived xenograft model of neuroblastoma. A: Curve of tumor volume change over time in patient-derived xenografts; B: Photographs of tumors in each group at the experimental endpoint; C: Statistical analysis of tumor weight in each group. Detailed Implementation

[0045] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. However, the embodiments of the present invention are not limited thereto. Various substitutions and modifications can be made based on ordinary technical knowledge and common practices in the art without departing from the above-described technical concept of the present invention, and all such substitutions and modifications should be included within the scope of the present invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in the art, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified can be purchased from legitimate channels as conventional products.

[0046] In the examples, the structure of the compounds was determined by nuclear magnetic resonance and high-resolution mass spectrometry. 1 H-NMR and 13 C-NMR was performed using a Bruker 500MHz NMR spectrometer with TMS as an internal standard. LC-HRMS was performed using an Agilent 1290-HPLC 6224 LC-MS system. Melting points were determined using a Buchi M565 melting point apparatus. Column chromatography was performed using 200-300 mesh silica gel.

[0047] Example 1

[0048]

[0049] Synthesis 1-1: 4.423 g of o-phthalaldehyde (33 mmol, 1.1 eq) and 2.251 g of glycine (30 mmol, 1 eq) were added to a 150 mL three-necked flask, dissolved in 60 mL of acetonitrile, and stirred under reflux overnight. LC-MS was used to monitor the reaction until complete. The mixture was filtered, and the filter cake was washed three times with 5 mL of acetonitrile. The solid was dried to obtain 4.083 g of a yellow-green solid, yield 71.2%. MS (ESI) + m / z measured value: 192.0646, calculated value: 192.0655 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.94(s,1H),7.71(d,J=7.5Hz,1H),7.64-7.59(m,2H),7.53-7.48(m,1H),4.52(s,2H),4.28(s,2H).

[0050] Synthesis of 1-2: 229 mg of compound 1-1 (1.2 mmol, 1.2 eq) and 569 mg of HBTU (1.5 mmol, 1.5 eq) were added to a 25 mL three-necked flask. 10 mL of DCM was added to dissolve the compound, and the mixture was stirred until homogeneous. Then, 349 μL of DIPEA (2 mmol, 2 eq) was added dropwise, and the mixture was stirred for 15 minutes. 229 mg of 3-amino-5-bromo-N-methylbenzamide (1 mmol, 1 eq) was added to the mixture, and the mixture was stirred for 2 hours. The reaction was then monitored by LC-MS to indicate completion. The mixture was diluted with 10 mL of DCM, washed three times with 10 mL of saturated NaHCO3 solution, and the organic phase was extracted and separated. The organic phase was then washed three times with 10 mL of saturated NH4Cl, extracted, and dried over anhydrous Na2SO4. The dried phase yielded 321 mg of a yellow solid, requiring no further purification, with a yield of 79.9%. MS (ESI) + m / z measured value: 402.0456, calculated value: 402.0448 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.89(s,1H),8.60(d,J=4.5Hz,1H),8.09(d,J=1.6Hz,1H),8.03(s,1H),7.75– 7.69(m,2H),7.63(d,J=4.2Hz,2H),7.54–7.47(m,1H),4.58(s,2H),4.45(s,2H),2.75(d,J=4.5Hz,3H).

[0051] Synthesis of A00: 281 mg of compounds 1-2 (0.7 mmol, 1 eq), 176 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.805 mmol, 1.15 eq), 342 mg of cesium carbonate (1.05 mmol, 1.5 eq), and 102 mg (0.14 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 150:1) to give 145 mg of white solid, yield 50.0%. mp 150.4-150.9℃. MS (ESI) + m / z measured value: 415.1762, calculated value: 415.1765 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.71(s,1H),8.85-8.76(m,2H),8.59(d,J=4.3Hz,1H),8.12(s,1H),8.02-7.94(m,2H),7.72 (d,J=7.5Hz,1H),7.67-7.61(m,3H),7.53-7.48(m,1H),4.59(s,2H),4.46(s,2H),2.78(d,J=4.4Hz,3H),2.50(s,3H).

[0052] Example 2

[0053] Referring to Example 1, glycine was replaced with D-alanine.

[0054]

[0055] Synthesis 2-1: 4.423 g of o-phthalaldehyde (33 mmol, 1.1 eq) and 2.673 g of D-alanine (30 mmol, 1 eq) were added to a 150 mL three-necked flask, dissolved in 60 mL of acetonitrile, and stirred under reflux overnight. LC-MS was used to monitor the reaction until complete. The mixture was filtered, and the filter cake was washed three times with 5 mL of acetonitrile. The solid was dried to obtain 4.566 g of a yellow-green solid, yield 75.0%. MS (ESI) + m / z measured value: 206.0810, calculated value: 206.0812 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.93(s,1H),7.71(d,J=7.5Hz,1H),7.64-7.60(m,2H),7.52-7.47(m,1H),4.52(s,2H),4.28(s,2H).

[0056] Synthesis of 2-2: 246 mg of compound 2-1 (1.2 mmol, 1.2 eq) and 569 mg of HBTU (1.5 mmol, 1.5 eq) were added to a 25 mL three-necked flask. 10 mL of LCM was added to dissolve the compound, and the mixture was stirred until homogeneous. Then, 349 μL of DIPEA (2 mmol, 2 eq) was added dropwise, and the mixture was stirred for 15 minutes. Next, 229 mg of 3-amino-5-bromo-N-methylbenzamide (1 mmol, 1 eq) was added, and the mixture was stirred for 2 hours. The reaction was then monitored by LC-MS to indicate completion. The mixture was diluted with 10 mL of LCM, washed three times with 10 mL of saturated NaHCO3 solution, and the organic phase was extracted and separated. The organic phase was then washed three times with 10 mL of saturated NH4Cl, extracted, and dried over anhydrous Na2SO4. The resulting 365 mg yellow solid was evaporated to dryness and used in the next step of the reaction without further purification. MS (ESI) + m / z measured value: 416.0612, calculated value: 416.0605 [M+H] + .

[0057] Synthesis of A01: 291 mg of compound 2-2 (0.7 mmol, 1 eq), 176 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.805 mmol, 1.15 eq), 342 mg of cesium carbonate (1.05 mmol, 1.5 eq), and 102 mg (0.14 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 150:1) to give 166 mg of white solid, yield 55.5%. mp 153.2-154.0℃. MS (ESI) + m / z measured value: 428.1854, calculated value: 428.1848 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ10.47(s,1H),8.47(dd,J=17.3,4.0Hz,2H),8.39(s,1H ),8.09(s,1H),7.83(s,1H),7.71(d,J=7.4Hz,1H),7.63(s,2H),7.52(d,J=18.2 Hz,2H),7.35(d,J=4.4Hz,1H),5.02(d,J=7.0Hz,1H),4.73(d,J=17.5Hz,1H),4. 62(d,J=17.5Hz,1H),2.77(d,J=3.7Hz,3H),2.26(s,3H),1.57(d,J=7.0Hz,3H).

[0058] Example 3

[0059] Referring to Example 1, compound 1-1 was replaced with phenylacetic acid.

[0060]

[0061] Synthesis of 3-1: 163 mg phenylacetic acid (1.2 mmol, 1.2 eq) and 569 mg HBTU (1.5 mmol, 1.2 eq) were prepared.

[0062] 1.5 eq) was added to a 25 mL three-necked flask, dissolved in 10 mL of DCM, and stirred until homogeneous. Then, 349 μL of DIPEA (2 mmol, 2 eq) was added dropwise, and the mixture was stirred for 15 minutes. Next, 229 mg of 3-amino-5-bromo-N-methylbenzamide (1 mmol, 1 eq) was added, and the mixture was stirred for 2 hours. The reaction was then monitored by LC-MS to indicate completion. The mixture was diluted with 10 mL of DCM, washed three times with 10 mL of saturated NaHCO3 solution, and the organic phase was extracted and separated. The organic phase was then washed three times with 10 mL of saturated NH4Cl, extracted, and dried over anhydrous Na2SO4. The resulting solid was evaporated to dryness to obtain 260 mg of a yellow solid, yielding 75.1% without further purification. MS (ESI) + m / z measured value: 347.0398, calculated value: 347.0390 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.51(s,1H),8.54(d,J=4.4Hz,1H),8.10(s,1H),7.94(s,1H),7.68( s,1H),7.32(t,J=6.4Hz,4H),7.25(dt,J=8.7,4.4Hz,1H),3.65(s,2H),2.76(d,J=4.5Hz,3H).

[0063] Synthesis of A02: 242 mg of compound 3-1 (0.7 mmol, 1 eq), 176 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.805 mmol, 1.15 eq), 342 mg of cesium carbonate (1.05 mmol, 1.5 eq), and 102 mg (0.14 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 180:1) to give 121 mg of white solid, with a yield of 48.1%. mp 205.1-205.5℃. MS (ESI) + m / z measured value: 360.1710, calculated value: 360.1707 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.46(s,1H),8.47(dd,J=14.7,4.5Hz,2H),8.40(s,1H),8.07(s,1H),7.85(s,1H),7.5 2(s,1H),7.33(dd,J=15.8,8.1Hz,5H),7.24(t,J=6.7Hz,1H),3.67(s,2H),2.78(d,J=4.3Hz,3H),2.26(s,3H).

[0064] Example 4

[0065] Referring to Example 3, phenylacetic acid was replaced with acetyl chloride.

[0066]

[0067] Synthesis of 4-1: 342 mg of 3-amino-5-bromo-N-methylbenzamide (1.5 mmol, 1 eq) was added to a 50 mL three-necked flask, and 15 mL of anhydrous DCM was added and stirred. 325 μL of triethylamine (2.25 mmol, 1.5 eq) was added and stirred for 5 minutes. 111 μL of acetyl chloride (1.575 mmol, 1.05 eq) was added dropwise to the system, and the reaction was stirred overnight. The reaction was monitored by TLC until completion. The mixture was filtered, and the filter cake was washed three times with 5 mL of DCM and dried to give 270 mg of white solid, yield 66.7%. MS (ESI) + m / z measured value: 271.0080, calculated value: 271.0077 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ10.30(s,1H),8.52(d,J=4.3Hz,1H),8.09(t,J=1.8Hz,1H) ,7.92(d,J=1.4Hz,1H),7.66(t,J=1.6Hz,1H),2.76(d,J=4.5Hz,3H),2.06(s,3H).

[0068] Synthesis of A03: 108 mg of compound 4-1 (0.4 mmol, 1 eq), 101 mg of pinacol 4-methylpyridine-3-boronate (0.46 mmol, 1.15 eq), 195 mg of cesium carbonate (0.6 mmol, 1.5 eq), and 58 mg (0.08 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged three times with N2. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was complete as monitored by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 40:1) to give 29 mg of white solid, yield 25.6%. mp 165.2-165.8 °C. MS(ESI + m / z measured value: 284.1404, calculated value: 284.1394 [M+H] + . 1 H NMR (500MHz, CDCl3) δ9.38 (s, 1H), 8.37 (d, J = 5.0Hz, 1H), 8.28 (s, 1H), 8.05 (s, 1H), 7.71 (s, 1H), 7.42 (s,1H),7.23(d,J=4.5Hz,1H),7.16(d,J=5.0Hz,1H),2.95(d,J=4.5Hz,3H),2.24(s,3H),2.18(s,3H).

[0069] Example 5

[0070]

[0071] Synthesis of 5-1: 228 mg of 3-amino-5-bromo-N-methylbenzamide (1 mmol, 1 eq) was added to a 50 mL three-necked flask, and 15 mL of anhydrous DCM was added with stirring. 300 μL of triethylamine (2 mmol, 2 eq) was added and stirred for 5 min. 119 μL of chloroacetyl chloride (1.5 mmol, 1.5 eq) was added dropwise to the system, and the reaction was stirred for 1 h. The reaction was monitored by TLC until completion. The mixture was filtered, and the filter cake was washed three times with 5 mL of DCM and dried to give 138 mg of gray solid, yield 45.4%. MS (ESI) + m / z measured value: 304.9688, calculated value: 304.9687 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.69(s,1H),8.57(d,J=4.5Hz,1H),8.07(t,J=2.0Hz,1H) ,7.96(d,J=1.5Hz,1H),7.73(t,J=1.5Hz,1H),4.29(s,2H),2.77(d,J=4.5Hz,3H).

[0072] Synthesis of 5-2: 121 mg of compound 5-1 (0.4 mmol, 1 eq) and 75 mg of N-Boc piperazine (0.4 mmol, 1 eq) were added to a 25 mL three-necked flask, and 4 mL of acetonitrile was added and stirred to dissolve. 83 mg of potassium carbonate (0.6 mmol, 1.5 eq) and 2 mg of potassium iodide (0.008 mmol, 0.02 eq) were added to the system, and the mixture was slowly heated to reflux and reacted overnight. The reaction was monitored by TLC until complete. The solvent was evaporated, the system was diluted with 10 mL of DCM, washed three times with 5 mL of 1 M hydrochloric acid, extracted, and the organic phase was collected. After drying in anhydrous Na₂SO₄, the organic phase was evaporated to dryness to give 64 mg of a yellow solid, with a yield of 35.2%. No further purification was required. MS (ESI) + m / z measured value: 455.1285, calculated value: 455.1289 [M+H] + . 1 H NMR (500MHz, CDCl3) δ10.91(s,1H),7.79(s,1H),7.63(s,1H),7.45(s,1H),7.40(s,1 H),4.34(s,2H),3.77-3.60(m,4H),2.94(d,J=4.0Hz,3H),2.18(s,4H),1.47(s,9H).

[0073] Synthesis of 5-3: 60 mg of compound 5-2 (0.1321 mmol, 1 eq), 33 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.1519 mmol, 1.15 eq), 65 mg of cesium carbonate (0.1982 mmol, 1.5 eq), and 19 mg (0.0264 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 4 mL of 1,4-dioxane and 1 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness to obtain a black solid, which was used for the next reaction without further purification.

[0074] Synthesis of A04: 0.1321 mmol of compound 5-3 was added to a 4 mL test tube, followed by 0.34 mL of THF and stirring to dissolve. 0.34 mL of a 4M solution of 1,4-dioxane hydrogen chloride was added dropwise, and the mixture was stirred for 2 h. The reaction was monitored by TLC until completion. The solid was filtered, washed three times with 1 mL of THF, and dried to obtain a brown solid. mp > 250 °C. MS (ESI) + m / z measured value: 368.2076, calculated value: 368.2081 [M+H] + . 1 H NMR (500MHz, DMSO) δ11.53(s,1H),10.05(s,2H),8.87(s,1H),8.85(d,J=5.9Hz,1H),8.79–8.72(m,1H),8.24(s,1H),8.0 8(d,J=5.8Hz,1H),7.94(s,1H),7.78(s,1H),4.34(s,2H),3.66(s,4H),3.47(s,4H),2.78(d,J=4.3Hz,3H),2.55(s,3H).

[0075] Example 6

[0076] Referring to Example 1, 4-methylpyridine-3-boronic acid pinacol ester was replaced with 4-methoxypyridine-3-boronic acid pinacol ester.

[0077]

[0078] Synthesis of A05: 281 mg of compounds 1-2 (0.7 mmol, 1 eq), 189 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.805 mmol, 1.15 eq), 342 mg of cesium carbonate (1.05 mmol, 1.5 eq), and 102 mg (0.14 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged three times with N2. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was complete as monitored by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 150:1) to give 233 mg of a white solid, yield 54.1%. mp>250 ​​°C. MS(ESI + m / z measured value: 431.1718, calculated value: 431.1714 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ10.46(s,1H),8.54(s,1H),8.48(s,1H),7.99-8.04(m,3H),7.80(s,1H),7.73(d,J=6. 5Hz,1H),7.63(s,2H),7.52(s,1H),6.95(d,J=8.5Hz,1H),4.61(s,2H),4.44(s,2H),3.90(s,3H),2.80(s,3H).

[0079] Example 7

[0080]

[0081] Synthesis of 7-1: 201 mg of compound 1-2 (0.5 mmol, 1 eq), 178 mg of N-tert-butoxycarbonyl-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (0.575 mmol, 1.15 eq), 244 mg of cesium carbonate (0.75 mmol, 1.5 eq), and 73 mg (0.1 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness, diluted with 10 mL of EtOAc and 10 mL of water, filtered with diatomaceous earth, and the filtrate was extracted and separated to obtain the organic phase. The organic phase was evaporated to dryness, recrystallized with a mixed solvent of MeOH / Et₂O, filtered, and the filter cake was dried to give 178 mg of a brown solid, with a yield of 70.6%. MS (ESI)+ m / z measured value: 505.2441, calculated value: 505.2446 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ10.51(s,1H),8.59–8.44(m,1H),7.93(s,1H),7.88(s,1H),7.72(d,J=7.5Hz,1H),7.65–7.61(m,3H),7.52(dd,J=8 .0,3.5Hz,1H),6.19(s,1H),4.59(s,2H),4.42(s,2H),4.00(s,2H),3.54(t,J=5.5Hz,2H),2.76(d,J=4.5Hz,3H),2.45(s,2H),1.42(s,9H).

[0082] Synthesis of A06: 33 mg of compound 7-1 (0.065 mmol, 1 eq) was added to a 4 mL test tube, followed by 0.16 mL of tetrahydrofuran. After stirring to dissolve, 0.16 mL of 4M hydrogen chloride solution in 1,4-dioxane was added dropwise. The reaction was stirred for 1 h, and the reaction was monitored by TLC until completion. The filter cake was collected, washed three times with 0.2 mL of tetrahydrofuran, and dried to give 19 mg of white solid, yield 66.0%. mp > 250 °C. MS (ESI) + m / z measured value: 405.1921, calculated value: 405.1921 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ10.81(s,1H),9.75(s,2H),8.67(d,J=3.0Hz,1H),8.00(s,1H),7.96(s,1H),7.75–7.68(m,2H),7.62(s ,2H),7.53–7.49(m,1H),6.24(s,1H),4.58(s,2H),4.45(s,2H),3.71(s,2H),3.26(s,2H),2.76(d,J=3.5Hz,3H),2.70(s,2H).

[0083] Example 8

[0084] Referring to Example 1, 4-methylpyridine-3-boronic acid pinacol ester was replaced with 2-methylphenylboronic acid pinacol ester.

[0085]

[0086] Synthesis of A07: 281 mg of compounds 1-2 (0.7 mmol, 1 eq), 189 mg of pinacol 2-methylphenylboronic acid (0.805 mmol, 1.15 eq), 342 mg of cesium carbonate (1.05 mmol, 1.5 eq), and 102 mg (0.14 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged three times with N2. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was complete as monitored by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 150:1) to give 233 mg of a white solid, yield 54.1%. mp>250 ​​°C. MS(ESI) + m / z measured value: 414.1825, calculated value: 414.1822 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ10.44(s,1H),8.47(d,J=4.5Hz,1H),8.03(s,1H),7.78(s,1H),7.73(d,J=7.5Hz,1H),7.63( d,J=4.0Hz,2H),7.55–7.47(m,2H),7.33–7.20(m,4H),4.59(s,2H),4.43(s,2H),2.77(d,J=4.5Hz,3H),2.23(s,3H).

[0087] Example 9

[0088]

[0089] Synthesis of A08: 263 mg of compound 1-1 (1.2 mmol, 1.2 eq) was added to a 25 mL three-necked flask, followed by 568 mg of HBTU (1.5 mmol, 1.5 eq). 4 mL of dichloromethane was added and stirred until dissolved. 273 μL of triethylamine was added dropwise, and the mixture was stirred for 15 minutes. 150 mg of 3-amino-N-methylbenzamide was added to the system, and the mixture was stirred for 2 hours. The reaction was monitored by LC-MS until complete. 5 mL of dichloromethane was added for dispersion and dilution. The filter cake was collected by vacuum filtration and washed three times with 4 mL of dichloromethane. Methanol was added to the filter cake until it could just be stirred and slurried. The filter cake was collected by vacuum filtration and dried to give 201 mg of a white solid, yield 62.2%. mp > 250 °C. MS (ESI) + m / z measured value: 324.1335, calculated value: 324.1343 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ10.35(s,1H),8.39(d,J=4.0Hz,1H),8.04(s,1H),7.73(t,J=7.0Hz,2H),7.63(d, J=4.0Hz,2H),7.53-7.50(m,,2H),7.38(d,J=8.0Hz,1H),4.59(s,2H),4.42(s,2H),2.76(d,J=4.5Hz,3H).

[0090] Example 10

[0091]

[0092] Synthesis of 10-1: 2.15 g of 3-amino-5-bromobenzoic acid (10 mmol, 1 eq), 1.31 mL of N-aminoethylmorpholine (10 mmol, 1 eq), and 4.56 g of HATU (12 mmol, 1.2 eq) were added to a 100 mL three-necked flask. 40 mL of dichloromethane was added and stirred to dissolve the mixture. 833 μL of DIPEA (20 mmol, 2 eq) was added dropwise, and the mixture was stirred for 5 hours. The reaction was monitored by TLC until completion. 40 mL of saturated NaHCO3 solution was added to the mixture, and the organic phase was extracted and separated. The mixture was cooled to 0 °C and recrystallized. The filter cake was collected by suction filtration, washed with cold DCM, and dried to give 2.859 g of a flesh-colored solid, yield 88.9%. MS (ESI) + m / z measured value: 328.0656, calculated value: 328.0655 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ8.48(s,1H),7.08(s,1H),7.01(s,1H),6.87(s,1H),5.63(s,2H),3.73(s,4H),3.48(s,2H),3.35(s,2H),2.99(s,4H).

[0093] Synthesis of 10-2: 573 mg of compound 1-1 (3 mmol, 1 eq) and 1.22 g of HATU (3.6 mmol, 1.2 eq) were added to a 50 mL three-necked flask. 15 mL of dichloromethane was added and stirred to dissolve. 250 μL of LDIPEA was added dropwise, and the mixture was stirred for 15 min. 981 mg of compound 10-1 (3 mmol, 1 eq) was added to the mixture, and the mixture was stirred overnight. The reaction was monitored by TLC until complete. The mixture was filtered, and the filter cake was washed three times with 15 mL of DCM and dried. The mixture was purified by column chromatography (DCM:MeOH = 30:1) to give 784 mg of a white solid, with a yield of 52.3%. MS (ESI) +m / z measured value: 501.1134, calculated value: 501.1132 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ10.55 (s, 1H), 8.80 (t, J = 5.4Hz, 1H), 8.07 (s, 1H), 8. 06(d,J=1.6Hz,1H),7.75(s,1H),7.73(d,J=7.6Hz,1H),7.64(d,J=1.5Hz,1H ),7.64(s,1H),7.55–7.47(m,1H),4.59(s,2H),4.43(s,2H),4.00(d,J=10.5 Hz,2H),3.74–3.58(m,4H),3.54(d,J=9.4Hz,2H),3.36(s,2H),3.13(s,2H).

[0094] Synthesis of A09: 102 mg of compound 10⁻² (0.216 mmol, 1 eq), 57 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.259 mmol, 1.15 eq), 106 mg of cesium carbonate (0.324 mmol, 1.5 eq), and 31 mg (0.043 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N₂ three times. A mixed solvent of 8 mL of 1,4-dioxane and 2 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness, diluted with 10 mL of DCM and 10 mL of water, extracted, and the organic phase was collected. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 20:1) to obtain 21 mg of orange solid, with a yield of 18.9%. mp146.9-147.1℃. MS(ESI) + m / z measured value: 514.2451, calculated value: 514.2451 [M+H] + . 1H NMR (500MHz, CDCl3) δ9.79 (s, 1H), 8.40 (d, J = 4.9Hz, 1H), 8.30 (s, 1H), 7.93 (s, 1H), 7.81(d,J=7.5Hz,1H),7.70(s,1H),7.55(t,J=7.4Hz,1H),7.46(s,1H),7.45(s,1H) ,7.43(s,1H),7.17(s,1H),7.13(d,J=4.9Hz,1H),4.63(s,2H),4.51(s,2H),3.66(s ,4H),3.49(dd,J=11.0,5.6Hz,2H),2.53(t,J=6.1Hz,2H),2.44(s,4H),2.20(s,3H).

[0095] Example 11

[0096]

[0097] Synthesis of 11-1: 2.15 g of 3-amino-5-bromobenzoic acid (10 mmol, 1 eq), 1.88 g of N-Boc piperazine (10.1 mmol, 1.01 eq), and 4.56 g of HATU (12 mmol, 1.2 eq) were added to a 100 mL three-necked flask. 40 mL of dichloromethane was added and stirred to dissolve the flask. 833 μL of DIPEA (20 mmol, 2 eq) was added dropwise, and the mixture was stirred for 3 hours. The reaction was monitored by TLC until completion. The filter cake was collected, washed three times with 10 mL of cold DCM, and dried to give 2.483 g of a white solid, yield 64.8%. MS (ESI) + m / z measured value: 384.0923, calculated value: 384.0918 [M+H] + . 1 H NMR (500MHz, CDCl3) δ6.85(t,J=1.9Hz,1H),6.80(t,J=1.5Hz,1H),6.58(dd,J=2.0,1.5Hz,1H),3.48(s,2H),2.79(s,8H),1.45(s,9H).

[0098] Synthesis of 11-2: 1.25 g of compound 1-1 (6.546 mmol, 1.01 eq) and 2.487 g of HATU (6.546 mmol, 1.01 eq) were added to a 50 mL three-necked flask. 25 mL of dichloromethane was added and stirred to dissolve. 543 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 2.483 g of compound 11-1 (6.482 mmol, 1 eq) was added to the mixture, and the mixture was stirred overnight. The reaction was monitored by TLC until complete. The mixture was washed three times with 25 mL of saturated NaHCO3 solution, and the organic phase was extracted and separated. The organic phase was then washed three times with 25 mL of saturated NH4Cl solution, extracted, and dried over anhydrous Na2SO4. The mixture was purified by column chromatography (DCM:MeOH = 50:1) to give 2.001 g of a white solid, yield 55.5%. MS (ESI) + m / z measured value: 557.1396, calculated value: 557.1394 [M+H] + . 1 HNMR(500MHz, CDCl3)δ9.25(s,1H),7.84(t,J=1.8Hz,1H),7.83–7.80(m,1H),7.59(td,J=7.6,1.1Hz,1H),7.48(d,J=1.4Hz ,2H),7.19(t,J=1.5Hz,1H),4.63(s,2H),4.45(s,2H),3.69(ddd,J=13.3,6.7,4.0Hz,3H),3.35-3.45(m,6H),1.45(s,9H).

[0099] Synthesis of 11-3: 222 mg of compound 11-2 (0.4 mmol, 1 eq), 105 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.48 mmol, 1.2 eq), 196 mg of cesium carbonate (0.6 mmol, 1.5 eq), and 58 mg (0.08 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 16 mL of 1,4-dioxane and 4 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness. The solution was diluted with 20 mL of DCM and 20 mL of water, extracted, and the organic phase was collected. The solution was dried over anhydrous Na2SO4. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 40:1) to give 122 mg of orange solid, yield 53.6%. MS(ESI + m / z measured value: 570.2705, calculated value: 570.2711 [M+H] + . 1H NMR(500MHz, CDCl3)δ9.51(s,1H),8.43(d,J=4.9Hz,1H),8.35(s,1H),7.83(d,J=7.4Hz,1H),7.71(s,1H),7.59–7.55(m,2H),7.46(t,J=6 .9Hz,2H),7.16(d,J=5.0Hz,1H),7.05(s,1H),4.63(s,2H),4.45(s,2H),3.59(d,J=110.6Hz,4H),3.41(s,3H),2.24(s,4H),1.45(s,9H).

[0100] Synthesis of A10: 117 mg of compound 11-3 was added to a 10 mL two-necked flask, and 2.5 mL of dichloromethane was added and stirred to dissolve. 0.4 mL of trifluoroacetic acid was added to the system, and the reaction was stirred for 1 h. The reaction was monitored by TLC until complete. The system was diluted with 5 mL of DCM, and 1 M NaOH solution was added dropwise to the system until pH = 12. The organic phase was extracted and separated, dried over anhydrous Na₂SO₄, and evaporated to dryness. No further purification was required to obtain 91 mg of the product as a yellow solid, yield 94.1%. mp > 250 °C. MS (ESI) + m / z measured value: 470.2182, calculated value: 470.2187 [M+H] + . 1 H NMR (500MHz, CDCl3) δ9.53(s,1H),8.42(d,J=4.9Hz,1H),8.35(s,1H),7.83(d,J=7.8Hz,1H),7.64(d,J=8.0Hz,2H),7.56(d,J=7.5Hz,1H),7.47(dd ,J=6.7,5.0Hz,2H),7.15(d,J=4.9Hz,1H),7.04(s,1H),4.61(s,2H),4.4 4(s,2H),3.96-3.26(m,4H),3.13-2.67(m,3H),2.24(s,1H),2.20(s,4H).

[0101] Example 12

[0102]

[0103] Synthesis of 12-1: 420 mg of compound 1-1 (2.2 mmol, 1.1 eq) and 988 mg of HATU (2.6 mmol, 1.3 eq) were added to a 50 mL three-necked flask. 20 mL of dichloromethane was added and stirred to dissolve. 400 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 430 mg of 3-amino-5-bromobenzoic acid (2 mmol, 1 eq) was added, and the mixture was stirred overnight. The reaction was monitored by TLC until complete. The mixture was filtered, and the filter cake was washed three times with 15 mL of DCM. The mixture was then slurried with a small amount of methanol to give 665 g of white solid, with a yield of 85.7%. MS (ESI) + m / z measured value: 389.0129, calculated value: 389.0132 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ13.37(s,1H),10.55(s,1H),8.14–8.13(m,2H),7.74– 7.72(m,2H),7.66–7.59(m,2H),7.55–7.49(m,1H),4.58(s,2H),4.42(s,2H).

[0104] Synthesis of 12-2: 388 mg of compound 12-1 (1 mmol, 1 eq), 252 mg of 4-methylpyridine-3-boronic acid pinacol ester (1.15 mmol, 1.15 eq), 489 mg of cesium carbonate (1.5 mmol, 1.5 eq), and 145 mg (0.2 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged with N2 three times. A mixed solvent of 30 mL of 1,4-dioxane and 7.5 mL of water was added to the system. After stirring until homogeneous, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed as monitored by LC-MS, the system was evaporated to dryness, diluted with 20 mL of DCM and 20 mL of water, and the aqueous phase was collected. The pH of the aqueous phase was adjusted to 2 with 1 M hydrochloric acid, and a large amount of solid precipitated. The solid was filtered, washed with 3 mL of 1 M hydrochloric acid, and dried to obtain 346 mg of white solid, with a yield of 86.3%. MS(ESI + m / z measured value: 402.1451, calculated value: 402.1449 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ13.20(s,1H),10.77(s,1H),8.60(d,J=5.5Hz,1H),8.57(s,1H),8.29(s,1H),7.96(t,J=2.0Hz,1H),7.72(d ,J=7.5Hz,1H),7.65(t,J=1.5Hz,1H),7.64–7.61(m,3H),7.51(ddd,J=8.0,6.5,4.0Hz,1H),4.59(s,2H),4.46(s,2H),2.35(s,3H).

[0105] Synthesis of A11: 100 mg of compound 12-2 (0.25 mmol, 1 eq) and 124 mg of HATU (0.325 mmol, 1.3 eq) were added to a 25 mL three-necked flask. 5 mL of dichloromethane was added and stirred to dissolve the compound. 50 μL of DIPEA was added dropwise and stirred for 15 min. 31 mg of cis-3,5-dimethylpiperidine (0.275 mmol, 1.1 eq) was added to the system, and the reaction was carried out for 2 hours. The reaction was monitored by TLC until completion. 5 mL of DCM was added for dilution. 10 mL of saturated NH4Cl solution was added to the system, and the mixture was extracted and separated. The organic phase was dried over anhydrous Na2SO4 and purified by column chromatography (DCM:MeOH = 40:1) to give 46 mg of a white solid, yield 37.1%. mp 145.1-145.7℃. MS (ESI) + m / z measured value: 497.2546, calculated value: 497.2547 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.45(s,1H),8.45(s,1H),8.37(s,1H),7.72(d,J=7.5Hz,1H), 7.66(d,J=11.5Hz,2H),7.62(d,J=3.0Hz,2H),7.51(dd,J=7.0,3.0Hz,1H),7.34(d,J=4. 5Hz,1H),7.07(s,1H),4.59(s,2H),4.44(s,3H),3.55(d,J=8.5Hz,1H),2.71–2.52(m,1 H),2.34–2.09(m,4H),1.77(d,J=12.0Hz,1H),1.58(d,J=5.5Hz,2H),0.99–0.59(m,7H).

[0106] Example 13

[0107]

[0108] Synthesis of 13-1: 2.94 g maleic anhydride (30 mmol, 1 eq) and 7.913 g anhydrous aluminum chloride were added to a 150 mL three-necked flask. 60 mL of anhydrous DCM was added and stirred until homogeneous. Under ice bath conditions, 3.242 g anisole (30 mmol, 1 eq) was added dropwise, and the reaction was stirred for 4 h. The reaction was monitored by TLC until completion. 120 mL of ice-cold 0.2 M hydrochloric acid was added dropwise, and the mixture was stirred vigorously. The mixture was then extracted, and the organic phase was evaporated to dryness. A small amount of methanol was used to redissolve the organic phase. The mixture was placed in a -20 °C refrigerator overnight, cooled, and recrystallized. The filter cake was collected and washed three times with 5 mL of cold methanol to give 4.88 g of a yellow solid, yield 79.0%. MS (ESI) + m / z measured value: 207.0655, calculated value: 207.0652 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ13.11(s,1H),8.06–8.02(m,2H),7.89(d,J=15.5Hz,1H),7.11–7.08(m,2H),6.65(d,J=15.5Hz,1H),3.87(s,3H).

[0109] Synthesis of A12: 201 mg of compound 13-1 (0.973 mmol, 1.05 eq) and 370 mg of HATU (0.973 mmol, 1.05 eq) were added to a 50 mL three-necked flask. 15 mL of DCM was added and stirred to dissolve the compound. 88 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 435 mg of compound A10 was then added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until completion. The mixture was purified by column chromatography (DCM:MeOH = 40:1) to give 380 mg of a yellow solid, yield 62.4%. mp 148.2-148.5. MS (ESI) + m / z measured value: 658.2657, calculated value: 658.2660 [M+H] + . 1H NMR (500MHz, CDCl3) δ9.68 (s, 1H), 8.42 (d, J = 5.0Hz, 1H), 8.31 (s, 1H), 8.02 (d, J = 8.5Hz, 2 H),7.95(d,J=15.0Hz,1H),7.81(d,J=7.5Hz,1H),7.73(s,1H),7.57(d,J=4.0Hz,1H),7.5 5(d,J=7.0Hz,1H),7.46(d,J=7.5Hz,2H),7.14(d,J=5.0Hz,1H),7.07(s,1H),6.96(d,J=9 .0Hz,2H),5.28(s,1H),4.64(s,2H),4.47(s,2H),3.88(s,3H),3.63(m,8H),2.22(s,3H).

[0110] Example 14

[0111] Referring to Example 13, compound 13-1 was replaced with acrylic acid.

[0112]

[0113] Synthesis of A13: 80 mg of compound A10 (0.1705 mmol, 1 eq), 15 mg of acrylic acid (0.205 mmol, 1.2 eq), and 97 mg of HATU (0.226 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 2 mL of DMA was added and stirred to dissolve the precipitate. 41 μL of LDIPEA was added to the system, and the mixture was stirred overnight. The reaction was monitored by LC-MS until completion. The system was then added dropwise to 20 mL of vigorously stirred water, resulting in the precipitation of a solid. The solid was filtered, washed three times with 5 mL of water, and dried to give 20 mg of a yellow solid, yielding 22.4%. mp 162.2–163.2 °C. MS (ESI) + m / z measured value: 524.2288, calculated value: 524.2293 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ10.59 (s, 1H), 8.50 (d, J = 4.9Hz, 1H), 8.46 (s, 1H), 7. 81–7.68(m,3H),7.62(d,J=3.8Hz,2H),7.51(td,J=7.8,4.4Hz,1H),7.45(d, J=5.0Hz,1H),7.17(s,1H),6.79(s,1H),6.13(dd,J=16.7,2.0Hz,1H),5.70( d,J=10.0Hz,1H),4.59(s,2H),4.45(s,2H),3.79–3.47(m,8H),2.32(s,3H).

[0114] Example 15

[0115] In Example 14, acrylic acid was replaced with chloroacetyl chloride.

[0116]

[0117] Synthesis of A14: 117 mg of compound A10 (0.25 mmol, 1 eq) and 24 μL of chloroacetyl chloride (0.3 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 2 mL of DCM was added and stirred to dissolve the compound. 109 μL of DIPEA was added to the system, and the mixture was stirred for 2 h. The reaction was monitored by TLC until completion. The mixture was washed with 2 mL of 1 M hydrochloric acid, extracted, and the organic phase was collected and dried over anhydrous Na2SO4. The organic phase was purified by column chromatography (DCM:MeOH = 30:1) to give 48 mg of a yellow solid, yield 35.2%. mp 194.7-195.0℃. MS (ESI) + m / z measured value: 546.1913, calculated value: 546.1903 [M+H] + . 1 H NMR (500MHz, DMSO) 1 H NMR(500MHz,DMSO)δ11.28–10.47(m,1H),9.11–8.73(m,1H),8.48-8.43(m, 1H),7.90(t,J=24.2Hz,1H),7.83–7.73(m,1H),7.72–7.67(m,1H),7.63(s,2 H),7.51(s,1H),7.40–7.23(m,1H),4.60(s,2H),4.47(d,J=9.3Hz,2H),4.3 8(t,J=5.1Hz,1H),3.65(dd,J=30.5,27.9Hz,6H),2.51(s,2H),1.99(s,5H).

[0118] Example 16

[0119] Referring to Example 14, acrylic acid was replaced with 5-norbornene-2-carboxylic acid.

[0120]

[0121] Synthesis of A15: 80 mg of compound A10 (0.1705 mmol, 1 eq), 28 mg of 5-norbornene-2-carboxylic acid (0.205 mmol, 1.2 eq), and 97 mg of HATU (0.226 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 2 mL of DCM was added and stirred to dissolve the compound. 41 μL of DIPEA was added to the system, and the mixture was stirred overnight. The reaction was monitored by LC-MS until completion. The system was diluted with 5 mL of DCM, washed with saturated NH4Cl solution, extracted, and the organic phase was dried over anhydrous Na2SO4. Column chromatography (DCM:MeOH = 50:1) was used for separation and purification, yielding 51 mg of a white solid (50.8% yield). mp 154.6-154.8℃. MS (ESI) + m / z measured value: 590.2758, calculated value: 590.2762 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ10.49(s,1H),8.45(d,J=4.9Hz,1H),8.39(s,1H),7.72(d,J=7.7Hz ,2H),7.68(s,1H),7.62(d,J=3.9Hz,2H),7.51(td,J=7.8,4.4Hz,1H),7.35(d,J=4.9Hz,1H) ,7.14(s,1H),6.15–6.04(m,1H),5.91–5.75(m,1H),4.59(s,2H),4.44(s,2H),3.46(d,J=8 7.1Hz,8H),2.82(d,J=17.0Hz,1H),2.28(s,3H),1.89(d,J=16.5Hz,1H),1.45–0.87(m,5H).

[0122] Example 17

[0123] Referring to Example 14, acrylic acid was replaced with 1-adamantaneacetic acid.

[0124]

[0125] Synthesis of A16: 94 mg of compound A10 (0.2 mmol, 1 eq), 47 mg of 1-adamantaneacetic acid (0.24 mmol, 1.2 eq), and 114 mg of HATU (0.3 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 4 mL of DCM was added and stirred to dissolve the compound. 50 μL of DIPEA was added to the system, and the mixture was stirred overnight. The reaction was monitored by LC-MS until completion. The system was purified by column chromatography (DCM:MeOH = 45:1) to give 107 mg of a white solid, yield 82.3%. mp 184.4-185.1℃. MS (ESI) + m / z measured value: 646.3387, calculated value: 646.3388 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ10.48(s,1H),8.45(d,J=5.0Hz,1H),8.39(s,1H),7.72(dd,J=4.6,2.8Hz,2H),7.67(t,J=1.6Hz,1H),7.64–7.60(m,2H),7. 35(d,J=5.0Hz,1H),7.14(s,1H),4.59(s,2H),4.44(s,2H),3.55(d,J=3.1 Hz,8H),2.28(s,3H),2.19–2.06(m,2H),1.89(s,3H),1.65–1.54(m,12H).

[0126] Example 18

[0127]

[0128] Synthesis of 18-1: 130 mg of compound 12-1 (0.335 mmol, 1 eq) and 190 mg of HATU (0.502 mmol, 1.5 eq) were added to a 25 mL three-necked flask. 4 mL of dichloromethane was added and stirred to dissolve. 175 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 62 mg of methyl 4-aminobutyrate hydrochloride (0.402 mmol, 1.2 eq) was added, and the mixture was stirred overnight. The reaction was monitored by TLC until complete. The mixture was filtered, and the filter cake was washed three times with 5 mL of DCM and dried to give 125 mg of a white solid, yield 76.7%. MS (ESI) + m / z measured value: 488.0817, calculated value: 488.0817 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ10.51(s,1H),8.59(t,J=5.5Hz,1H),8.08(s,1H),7.94(s,1H),7.73(d,J=6.5Hz,2H),7.65–7.60(m,2H),7. 55–7.49(m,1H),4.58(s,2H),4.42(s,2H),3.58(s,3H),3.25(dd,J=12.5,6.5Hz,2H),2.36(t,J=7.5Hz,2H),1.77(p,J=7.0Hz,2H).

[0129] Synthesis of 18-2: 168 mg of compound 18-1 (0.345 mmol, 1 eq) was added to a 25 mL three-necked flask, and 4 mL of THF was added and stirred to dissolve. 29 mg (0.69 mmol, 2 eq) of lithium hydroxide monohydrate was dissolved in 2 mL of water and added dropwise to the reaction system. The mixture was stirred for 5 h, and the reaction was monitored by LC-MS until completion. 1 M hydrochloric acid was added dropwise to the system until no more solid precipitated. The mixture was filtered, and the filter cake was washed three times with 1 mL of 1 M hydrochloric acid. The filter cake was dried to give 114 mg of an off-white solid, with a yield of 69.9%. MS (ESI) + m / z measured value: 474.0674, calculated value: 474.0669 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.09(s,1H),10.53(s,1H),8.58(t,J=5.4Hz,1H),8.09(s,1H),7.94(s,1H),7.74–7.71(m,2H),7.63(d,J= 3.1Hz,2H),7.53–7.50(m,1H),4.58(s,2H),4.42(s,2H),3.25(dd,J=12.6,6.6Hz,2H),2.27(t,J=7.4Hz,2H),1.74(p,J=7.2Hz,2H).

[0130] Synthesis of 18-3: 111 mg of compound 18-2 (0.234 mmol, 1 eq) and 67 mg of HATU (0.258 mmol, 1.25 eq) were added to a 25 mL three-necked flask. 2 mL of LDMF was added and stirred to dissolve. 61 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 67 mg of lenalidomide (0.258 mmol, 1.1 eq) was added, and the mixture was stirred overnight. The reaction was monitored by LC-MS until complete. The mixture was then added dropwise to 20 mL of vigorously stirred ice water, resulting in the precipitation of a large amount of solid. The solid was filtered, washed three times with 5 mL of water, dried, and used as feed for the next reaction without further purification. MS (ESI)+ m / z measured value: 715.1514, calculated value: 715.1511 [M+H] + .

[0131] Synthesis of A17: 71 mg of compound 18-3 (0.1 mmol, 1 eq), 25 mg of 4-methylpyridine-3-boronic acid pinacol ester (0.115 mmol, 1.15 eq), 49 mg of cesium carbonate (0.15 mmol, 1.5 eq), and 15 mg (0.02 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 10 mL two-necked flask. The mixture was purged three times with N2. A mixed solvent of 3 mL of 1,4-dioxane and 0.75 mL of water was added, and the mixture was stirred until homogeneous. The temperature was slowly increased to 65 °C and reacted for 1 h. After the reaction was complete as monitored by LC-MS, the mixture was evaporated to dryness, diluted with 2 mL of DCM and 2 mL of water, and the organic phase was collected. The crude product was purified by separation with DCM:MeOH = 40:1 to give 40 mg of white solid, yield 55.0%. MS (ESI) + m / z measured value: 728.2830, calculated value: 728.2827 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ11.00-10.48(m,2H),8.57(t,J=5.8Hz,1H),8.45(d,J=5.0Hz,1H),8.41(d,J=2.6Hz,1H),8.1 0–8.05(m,1H),7.84-7.78(m,2H),7.72(d,J=7.5Hz,1H),7.63(d,J=4.2Hz,2H),7.58(t,J=4.4Hz,1H),7.53–7.39(m,3 H),7.35(d,J=4.9Hz,1H),4.59(s,2H),4.43(s,2H),4.42–4.27(m,2H),3.31–3.22(m,2H),2.96–2.90(m,1H),2.89–2 .75(m,1H),2.65–2.56(m,1H),2.42(t,J=7.4Hz,1H),2.37–2.31(m,1H),2.26(s,3H),2.11–1.82(m,3H),1.23(s,1H).

[0132] Example 19

[0133] In Example 13, maleic anhydride was replaced with succinic anhydride.

[0134]

[0135] Synthesis of 19-1: 1.001 g succinic anhydride (10 mmol, 1 eq) and 2.902 g anhydrous aluminum chloride (22 mmol, 2.2 eq) were added to a 150 mL three-necked flask. 60 mL of anhydrous DCM was added and stirred thoroughly. Under ice bath conditions, 1.195 mL of anisole (11 mmol, 1.1 eq) was added dropwise, and the reaction was stirred for 4 h. The reaction was monitored by TLC until completion. 120 mL of ice-cold 0.2 M hydrochloric acid was added dropwise, and the mixture was stirred vigorously. The organic phase was then extracted and evaporated to dryness. A small amount of methanol was used to redissolve the solid. The mixture was placed in a -20 °C refrigerator overnight, cooled, and recrystallized. The filter cake was collected and washed three times with 5 mL of cold methanol to give 1.54 g of a yellow solid, yield 74.0%. MS (ESI) + m / z measured value: 209.0801, calculated value: 209.0809 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.08(s,1H),7.99–7.93(m,2H),7.07–7.02(m,2H),3.84(s,3H),3.21–3.17(m,2H),2.58–2.53(m,2H).

[0136] Synthesis of A18: 60 mg of compound A10 (0.128 mmol, 1 eq), 32 mg of compound 19-1 (0.1537 mmol, 1.2 eq), and 73 mg of HATU (0.192 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 4 mL of DCM was added and stirred to dissolve the mixture. 20 μL of DIPEA was added to the system, and the mixture was stirred for 4 h. The reaction was monitored by LC-MS until completion. The system was purified by column chromatography (DCM:MeOH = 40:1) to give 65 mg of a white solid, yield 65.7%. mp 175.9-176.8℃. MS (ESI) + m / z measured value: 660.2819, calculated value: 660.2817 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ10.53(s,1H),8.45(d,J=5.2Hz,1H),8.40(s,1H),7.95(d,J=8.8 Hz,2H),7.83–7.70(m,2H),7.68(s,1H),7.62(d,J=3.9Hz,2H),7.51(td,J=7.9,4.4Hz,1H ),7.35(d,J=4.8Hz,1H),7.16(s,1H),7.03(d,J=8.8Hz,2H),4.59(s,2H),4.45(s,2H),3. 83(s,3H),3.57(t,J=53.5Hz,8H),3.18(t,J=5.8Hz,2H),2.77–2.60(m,2H),2.29(s,3H).

[0137] Example 20

[0138]

[0139] Synthesis of 20-1: 573 mg of compound 1-1 (3 mmol, 1 eq) and 1.368 g of HATU (3.6 mmol, 1.2 eq) were added to a 50 mL three-necked flask. 20 mL of dichloromethane was added and stirred to dissolve. 500 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 453 mg of 3-aminobenzoic acid (3.3 mmol, 1.1 eq) was added, and the mixture was stirred overnight. The reaction was monitored by TLC until complete. The mixture was filtered, and the filter cake was washed three times with 15 mL of LCM and slurried with a small amount of methanol to give 690 g of white solid, yield 74.2%. MS (ESI) + m / z measured value: 311.1021, calculated value: 311.1027 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.95(s,1H),10.39(s,1H),8.23(s,1H),7.81(dd,J=8.0,1.0Hz,1H),7.73(d ,J=7.5Hz,1H),7.67–7.59(m,3H),7.55–7.49(m,1H),7.45(d,J=8.0Hz,1H),4.59(s,2H),4.42(s,2H).

[0140] Synthesis of 20-2: 310 mg of compound 20-1 (1 mmol, 1 eq), 224 mg of N-Boc piperazine (1.2 mmol, 1.2 eq), and 456 mg of HATU (1.2 mmol, 1.2 eq) were added to a 25 mL three-necked flask. 5 mL of dichloromethane was added and stirred to dissolve the compound. 167 μL of DIPEA (2 mmol, 2 eq) was added dropwise and the mixture was stirred for 3 hours. The reaction was monitored by TLC until completion. 5 mL of LCM was added for dilution. 10 mL of saturated NH4Cl solution was added to the mixture, and the organic phase was extracted and separated. The mixture was then washed with 10 mL of saturated NaHCO3, extracted, and separated again. The organic phase was dried over anhydrous Na2SO4. The crude product was purified by column chromatography (100:1) to give 321 mg of a white solid, with a yield of 67.2%. MS (ESI) + m / z measured value: 479.2291, calculated value: 479.2289 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.92 (s, 1H), 7.87 (d, J = 7.5Hz, 1H), 7.63 (s, 1H), 7.59 (td, J = 7.5, 1.0Hz, 1H), 7.54 (d, J = 8.0Hz, 1H), 7.50 (dd, J = 9. 5,8.0Hz,2H),7.32(t,J=8.0Hz,1H),7.10(d,J=7.5Hz,1H),4.61(s,2H),4.41(s,2H),3.76–3.63(m,2H),3.54–3.32(m,6H),1.46(s,9H).

[0141] Synthesis of 20-3: 321 mg of compound 20-2 (0.671 mmol) was added to a 25 mL two-necked flask, and 5 mL of dichloromethane was added and stirred to dissolve. 1.332 mL of trifluoroacetic acid was added to the system, and the reaction was stirred for 1 h. The reaction was monitored by TLC until complete. The system was diluted with 5 mL of DCM, and 1 M NaOH solution was added dropwise to the system until pH = 12. The organic phase was extracted and separated, dried over anhydrous Na₂SO₄, and evaporated to dryness. No further purification was required to obtain 242 mg of the product as a white solid, with a yield of 95.4%. MS (ESI) + m / z measured value: 379.1760, calculated value: 379.1765 [M+H] + . 1H NMR(500MHz, CDCl3)δ8.90(s,1H),7.88(d,J=7.5Hz,1H),7.60(dd,J=7.5,1.0Hz,1H),7.58–7.55(m,2H),7.50(dd,J=12.0 ,7.5Hz,2H),7.31(t,J=8.0Hz,1H),7.10(d,J=7.5Hz,1H),4.61(s,2H),4.40(s,2H),2.99–2.87(m,2H),2.80–2.90(m,6H).

[0142] Synthesis of A19: 48 mg of compound 13-1 (0.24 mmol, 1.2 eq) and 114 mg of HATU (0.3 mmol, 1.5 eq) were added to a 25 mL three-necked flask. 4 mL of DCM was added and stirred to dissolve the compound. 70 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 76 mg of compound 20-3 was then added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until complete. The mixture was purified by column chromatography (DCM:MeOH = 40:1) to give 27 mg of a yellow solid, yield 23.9%. mp 97.9–98.8 °C. MS (ESI) + m / z measured value: 567.2242, calculated value: 567.2238 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.89 (s, 1H), 8.04 (d, J = 9.0Hz, 2H), 7.97 (d, J = 15.0Hz ,1H),7.89(d,J=7.5Hz,1H),7.70(s,1H),7.60(td,J=7.5,1.0Hz,1H),7.53- 7.49(m,3H),7.35(t,J=8.0Hz,1H),7.14(d,J=7.5Hz,1H),6.98(d,J=9.0Hz, 2H),4.62(s,2H),4.40(s,2H),3.89(s,3H),3.90-3,50(m,8H),2.80(s,1H).

[0143] Example 21

[0144]

[0145] Synthesis of 21-1: 1.742 g of o-phthalaldehyde (13 mmol, 1.3 eq) was added to a 50 mL three-necked flask, followed by 5 mL of ethyl acetate and 2.5 mL of acetic acid. The mixture was stirred until homogeneous and then slowly heated to 45 °C. 2.15 g of 3-amino-5-bromo-benzoic acid (10 mmol, 1 eq) was dissolved in 5 mL of ethyl acetate and added dropwise. The mixture was stirred overnight, and the reaction was monitored by LC-MS until completion. The mixture was filtered, and the filter cake was washed three times with 5 mL of ethyl acetate and dried to give 2.45 g of a brown solid, yield 74.0%. MS (ESI) + m / z measured value: 331.9913, calculated value: 331.9917 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ13.46 (s, 1H), 8.50–8.46 (m, 1H), 8.43 (t, J = 2.0Hz, 1H), 7.82 (dd, J = 5. 8,4.5Hz,2H),7.71(ddd,J=21.2,10.8,4.2Hz,2H),7.56(dd,J=10.9,3.8Hz,1H),5.11(s,2H).

[0146] Synthesis of 21-2: 1.986 g of compound 21-1 (6 mmol, 1 eq), 1.34 g of N-Boc piperazine (7.2 mmol, 1.2 eq), and 2.729 g of HBTU (7.2 mmol, 1.2 eq) were added to a 150 mL three-necked flask. 40 mL of dichloromethane was added and stirred to dissolve the mixture. 1.8 mL of triethylamine was added dropwise, and the mixture was stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was filtered, and the filter cake was slurried with a small amount of methanol. The filtered cake was dried to obtain 2.707 g of a white solid, yield 90.4%. MS (ESI) + m / z measured value: 999.2290, calculated value: 999.2287 [2M+H] + . 1 H NMR (500MHz, CDCl3) δ8.04 (dd, J=4.3, 3.0Hz, 2H), 7.92 (d, J=8.0Hz, 1H), 7.64 (td, J=7.5, 1.0Hz,1H),7.56–7.51(m,2H),7.35(s,1H),4.86(s,2H),3.78–3.44(m,8H),1.47(s,9H).

[0147] Synthesis of 21-3: 2.707 g of compound 20-2 (5.42 mmol) was added to a 25 mL two-necked flask, and 8.1 mL of dichloromethane was added and stirred to dissolve. 2.7 mL of trifluoroacetic acid was added to the system, and the reaction was stirred for 1 h. The reaction was monitored by TLC until complete. The system was evaporated to dryness, and the remaining oily liquid was added dropwise to 20 mL of vigorously stirred 1 M NaOH solution, precipitating a large amount of solid. The solid was filtered, washed three times with 5 mL of cold water, and dried to give 2127 mg of white solid, yield 98.4%. MS (ESI) + m / z measured value: 400.0653, calculated value: 400.0653 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ8.28(t,J=1.9Hz,1H),7.96(s,1H),7.81(d,J=7.6Hz,1H),7.70(dt,J=18.0,7. 4Hz,2H),7.57(t,J=7.4Hz,1H),7.44(s,1H),5.08(s,2H),3.76(s,2H),3.55–3.34(m,3H),3.04(s,4H).

[0148] Synthesis of A20: 96 mg of compound 13-1 (0.48 mmol, 1.2 eq) and 228 mg of HATU (0.6 mmol, 1.5 eq) were added to a 25 mL three-necked flask. 5 mL of DCM was added and stirred to dissolve the compound. 140 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 160 mg of compound 20-3 (0.4 mmol, 1 eq) was added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until completion. The mixture was purified by column chromatography (DCM:MeOH = 60:1) to give 96 mg of a yellow solid, yield 40.9%. mp 140.5–141.5 °C. MS (ESI) + m / z measured value: 588.1129, calculated value: 588.1129 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.14(s,1H),8.06–7.97(m,3H),7.95(t,J=7.4Hz,1H),7.90(d,J=7.3Hz,1H),7.63(t,J=7.2H z,1H),7.56–7.40(m,3H),7.36(d,J=10.7Hz,1H),6.99–6.94(m,2H),4.86(s,2H),3.88(s,3H),3.87-3.58(m,8H).

[0149] Example 22

[0150] Referring to Example 21, 3-amino-5-bromo-benzoic acid was replaced with m-aminobenzoic acid.

[0151]

[0152] Synthesis of 22-1: 536 g of o-phthalaldehyde (4 mmol, 1 eq) was added to a 25 mL three-necked flask, followed by 2 mL of ethyl acetate and 1 mL of acetic acid. The mixture was stirred until homogeneous. 549 mg of m-aminobenzoic acid (4 mmol, 1 eq) was dissolved in 2 mL of ethyl acetate and added dropwise. The mixture was stirred for 1 h, and the reaction was monitored by TLC until completion. The mixture was filtered, and the filter cake was washed three times with 5 mL of ethyl acetate. The mixture was recrystallized using a mixed solvent of MeOH and Et₂O, filtered again, and the filter cake was dried to give 513 mg of a brown solid, with a yield of 50.7%. MS (ESI) + m / z measured value: 254.0810, calculated value: 254.0812 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ13.07(s,1H),8.53(s,1H),8.20–8.12(m,1H),7.81(d,J=7. 5Hz,1H),7.75(d,J=7.5Hz,1H),7.72–7.67(m,2H),7.60–7.53(m,2H),5.09(s,2H).

[0153] Synthesis of 22-2:

[0154] 430 g of compound 22-1 (1.7 mmol, 1 eq), 380 g of N-Boc piperazine (2.04 mmol, 1.2 eq), and 775 mg of HBTU (2.04 mmol, 1.2 eq) were added to a 50 mL three-necked flask. 17 mL of dichloromethane was added and stirred to dissolve the compound. 0.51 mL of DIPEA was added dropwise, and the mixture was stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was evaporated to dryness, and the crude product was recrystallized from a cooled, hot, saturated methanol solution. The product was filtered while cold, and the filter cake was dried to give 560 g of an orange solid, with a yield of 78.2%. MS (ESI) + m / z measured value: 843.4078, calculated value: 843.4077 [2M+H] + . 1 H NMR(500MHz,DMSO-d6)δ7.98(d,J=6.5Hz,2H),7.80(d,J=7.5Hz,1H),7.72–7.66(m,2H) ,7.58–7.50(m,2H),7.21(d,J=7.5Hz,1H),5.07(s,2H),3.78–3.34(m,8H),1.41(s,9H).

[0155] Synthesis of 22-3:

[0156] 522 mg (1.24 mmol) of compound 22-2 was added to a 25 mL two-necked flask, and 6.2 mL of dichloromethane was added and stirred to dissolve. 1.24 mL of trifluoroacetic acid was added to the system, and the reaction was stirred for 1 h. The reaction was monitored by TLC until complete. The system was evaporated to dryness, and the remaining oily liquid was added dropwise to 20 mL of vigorously stirred 1 M NaOH solution, resulting in the precipitation of a large amount of solid. The solid was filtered, and the filter cake was washed three times with 5 mL of cold water and dried to obtain 361 mg of orange solid, with a yield of 90.7%. MS (ESI) + m / z measured value: 322.1553, calculated value: 322.1550 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ7.99(s,1H),7.93(d,J=8.0Hz,1H),7.80(d,J=7.5Hz,1H),7.71–7.69(m,2H),7.55(t,J=7 .0Hz,1H),7.51(t,J=8.0Hz,1H),7.17(d,J=7.5Hz,1H),5.07(s,2H),3.56(s,2H),3.28(s,2H),2.77–2.62(m,5H).

[0157] Synthesis of A21: 52 mg of compound 13-1 (0.25 mmol, 1 eq) and 114 mg of HBTU (0.3 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 2.5 mL of LDCM was added and stirred to dissolve. 70 μL of LDIPEA was added dropwise, and the mixture was stirred for 15 min. 80 mg of compound 22-3 (0.25 mmol, 1 eq) was added to the mixture, and the mixture was stirred for 4 h. The reaction was monitored by TLC until complete. The mixture was recrystallized from a cooled, hot, saturated methanol solution. The filter cake was collected by suction filtration, dried, and yielded 63 mg of a flesh-colored solid, with a yield of 49.6%. mp 193.4–193.8 °C. MS (ESI) + m / z measured value: 510.2026, calculated value: 510.2024 [M+H] + . 1H NMR(500MHz, DMSO-d6)δ8.03(s,3H),7.99(d,J=8.0Hz,1H),7.85–7.75(m,2H),7.74–7.65(m,2H),7.54(dd,J=15.0,7. 0Hz,2H),7.49–7.35(m,1H),7.24(d,J=7.5Hz,1H),7.09(d,J=7.0Hz,2H),5.07(s,2H),3.86(s,3H),3.79–3.41(m,8H).

[0158] Example 23

[0159] Referring to Example 22, m-aminobenzoic acid was replaced with 5-aminonicotinic acid.

[0160]

[0161] Synthesis of 23-1: 536 g of o-phthalaldehyde (4 mmol, 1 eq) was added to a 25 mL three-necked flask, followed by 2 mL of ethyl acetate and 1 mL of acetic acid. The mixture was stirred until homogeneous. 549 mg of 5-aminonicotinic acid (4 mmol, 1 eq) was dissolved in 2 mL of ethyl acetate and added dropwise. The mixture was stirred for 1 h, and the reaction was monitored by TLC until completion. The mixture was filtered, and the filter cake was washed three times with 5 mL of ethyl acetate. The mixture was recrystallized using a mixed solvent of MeOH and Et₂O, filtered again, and the filter cake was dried to give 870 mg of a pale yellow solid, with a yield of 85.6%. MS (ESI) + m / z measured value: 255.0766, calculated value: 255.0764 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ13.53(s,1H),9.27(d,J=2.5Hz,1H),8.92–8.82(m,2H),7.8 3(d,J=7.5Hz,1H),7.71(q,J=7.5Hz,2H),7.57(dt,J=7.5,0.5Hz,1H),5.15(s,2H).

[0162] Synthesis of 23-2: 508 mg of compound 23-1 (2 mmol, 1 eq), 447 g of N-Boc piperazine (2.4 mmol, 1.2 eq), and 910 mg of HBTU (2.4 mmol, 1.2 eq) were added to a 50 mL three-necked flask. 20 mL of dichloromethane was added and stirred to dissolve the compound. 0.6 mL of DIPEA was added dropwise, and the mixture was stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was evaporated to dryness. The crude product was recrystallized from a cooled, hot, saturated methanol solution. The residue was filtered while cold, and the filter cake was dried to give 541 g of a white-yellow solid, with a yield of 64.1%. MS (ESI) +m / z measured value: 423.2028, calculated value: 423.2027 [M+H] + . 1 H NMR(500MHz, CDCl3)δ9.00(d,J=2.0Hz,1H),8.65–8.58(m,1H),8.48(d,J=1.0Hz,1H),7.93(d,J=7.5Hz, 1H),7.66(td,J=7.5,1.0Hz,1H),7.57-7.53(m,2H),4.93(s,2H),3.77-3.71(m,2H),3.55-3.48(m,6H).

[0163] Synthesis of 23-3: 483 mg of compound 23-2 (1.15 mmol) was added to a 25 mL two-necked flask, and 5.5 mL of dichloromethane was added and stirred to dissolve. 1.2 mL of trifluoroacetic acid was added to the system, and the reaction was stirred for 1 h. The reaction was monitored by TLC until complete. The system was evaporated to dryness, and the remaining oily liquid was added dropwise to 20 mL of vigorously stirred 1 M NaOH solution, precipitating a large amount of solid. The solid was filtered, and the filter cake was washed three times with 5 mL of cold water and dried to give 288 mg of a yellow solid, with a yield of 78.0%. MS (ESI) + m / z measured value: 323.1501, calculated value: 323.1503 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ7.99(s,1H),7.93(d,J=9.0Hz,1H),7.80(d,J=7.5Hz,1H),7.72–7.66(m,2H),7.55(t,J=7.0H z,1H),7.51(t,J=8.0Hz,1H),7.17(d,J=7.5Hz,1H),5.07(s,2H),3.56(s,2H),3.28(s,2H),2.74(s,2H),2.65(s,2H).

[0164] Synthesis of A22: 74 mg of compound 13-1 (0.36 mmol, 1.2 eq) and 171 mg of HBTU (0.45 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 3 mL of LCM was added and stirred to dissolve the compound. 90 μL of LIPEA was added dropwise, and the mixture was stirred for 15 min. 97 mg of compound 23-3 (0.3 mmol, 1 eq) was added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until complete. The filter cake was collected, washed with 2 mL of LCM, then washed with 2 mL of saturated NH4Cl solution, saturated NaHCO3 solution, and water, respectively. After drying, 63 mg of a yellow solid was obtained, with a yield of 41.1%. mp 246.8–247.0 °C. MS (ESI) +m / z measured value: 511.1982, calculated value: 511.1976 [M+H] + . 1 HNMR(500MHz,DMSO-d6)δ9.20(d,J=2.0Hz,1H),8.54–8.37(m,2H),8.04(d,J=6.0Hz,2H),7.82(t,J=11.5Hz,2H),7.76–7.67(m,2H ),7.58(t,J=7.0Hz,1H),7.49–7.38(m,1H),7.09(d,J=7.5Hz,2H),5.13(s,2H),3.86(s,3H),3.80–3.60(m,6H),3.49–3.47(m,2H).

[0165] Example 24

[0166]

[0167] Synthesis of A23: 59 mg of compound A20 (0.1 mmol, 1 eq), 25 mg of 4-methylpyridine-3-boronate pinacol ester (0.115 mmol, 1.15 eq), 49 mg of cesium carbonate (0.15 mmol, 1.5 eq), and 15 mg (0.02 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged three times with N2. A mixed solvent of 4 mL of 1,4-dioxane and 1 mL of water was added to the system. After stirring, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was completed by LC-MS, the system was evaporated to dryness. The solution was diluted with 5 mL of DCM and 5 mL of water, extracted, and the organic phase was collected and dried over Na2SO4. The crude product was purified by column chromatography (DCM:MeOH = 50:1) to give 20 mg of white solid, yield 33.5%. mp113.0-113.3℃. MS(ESI) + m / z measured value: 601.2440, calculated value: 601.2446 [M+H] + . 1H NMR (500MHz, CDCl3) δ8.47(s,2H),8.12(s,1H),8.01(d,J=8.5Hz,2H),7.96(d,J= 15.0Hz,1H),7.90(d,J=7.5Hz,2H),7.61(dd,J=11.0,4.0Hz,1H),7.52(d,J=7.5H z,2H),7.46(dd,J=6.5,3.5Hz,1H),7.22(d,J=5.0Hz,1H),7.18(d,J=9.5Hz,1H), 6.95(d,J=9.0Hz,2H),4.91(s,2H),3.86-3.66(m,8H),2.35(s,3H),1.23(s,3H).

[0168] Example 25

[0169]

[0170] Synthesis of 25-1: 1.88 g of p-trifluoromethylacetophenone (10 mmol, 1 eq) and 920 mg of glyoxylic acid monohydrate (10 mmol, 1 eq) were added to a 50 mL three-necked flask. 20 mL of acetic acid was added, and the mixture was stirred and refluxed overnight. The reaction was monitored by LC-MS until complete. Acetic acid was evaporated to dryness, and the crude product was purified by column chromatography (DCM:MeOH:AcOH = 100:1:0.1) to give 1.37 g of a white-yellow solid, yield 56.1%. MS (ESI) + m / z measured value: 245.0420, calculated value: 245.0420 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.11(d,J=8.1Hz,2H),7.96(d,J=15.6Hz,1H),7.80(d,J=8.3Hz,2H),6.94(d,J=15.6Hz,1H).

[0171] Synthesis of A24: 44 mg of compound 25-1 (0.18 mmol, 1.2 eq) and 85 mg of HATU (0.225 mmol, 1.5 eq) were added to a 25 mL three-necked flask. 5 mL of DCM was added and stirred to dissolve the compound. 50 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 60 mg of compound 21-3 (0.15 mmol, 1 eq) was added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until completion. The mixture was purified by column chromatography (DCM:MeOH = 130:1) to give 38 mg of a yellow solid, yield 40.5%. mp 101.1–101.9 °C. MS (ESI) +m / z measured value: 626.0896, calculated value: 626.0897 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.19(s,1H),8.14(d,J=7.9Hz,2H),7.94(dd,J=19.6,11.3Hz,3H),7.78(d,J= 8.2Hz,2H),7.65(t,J=7.5Hz,1H),7.56–7.53(m,3H),7.39(s,1H),4.87(s,2H),3.83-3.71(m,8H).

[0172] Example 26

[0173]

[0174] Synthesis of 26-1: 99 mg of compound 21-1 (0.3 mmol, 1 eq), 62 mg of N-Boc butanediamine (0.33 mmol, 1.1 eq), and 136 mg of HATU (0.36 mmol, 1.2 eq) were added to a 10 mL two-necked flask. 3 mL of dichloromethane was added and stirred to dissolve the compound. 150 μL of triethylamine was added dropwise, and the mixture was stirred for 2 hours. The reaction was monitored by TLC until completion. The mixture was diluted with 3 mL of DCM, washed three times with 5 mL of saturated sodium bicarbonate solution, extracted, and the organic phase was washed three times with 5 mL of 1 M hydrochloric acid. The organic phase was dried over anhydrous Na₂SO₄ and allowed to proceed to the next step without further purification. MS (ESI) + m / z measured value: 1003.2598, calculated value: 1003.2599 [2M+H] + .

[0175] Synthesis of 26-2: 0.3 mmol of compound 26-1 was added to a 10 mL two-necked flask, followed by 0.75 mL of tetrahydrofuran with stirring to dissolve. 0.75 mL of a 4M solution of 1,4-dioxane hydrogen chloride was added dropwise to the system, and the reaction was stirred for 2 h. The reaction was monitored by LC-MS until completion. The mixture was filtered, and the filter cake was washed three times with 2 mL of tetrahydrofuran and dried to give 101 mg of a white solid, yield 77.0%. MS (ESI) + m / z measured value: 402.0817, calculated value: 402.0822 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ8.84(t,J=5.6Hz,1H),8.48(t,J=1.9Hz,1H),8.30–8.24(m,1H),7.99(s,3H),7.84(t,J=1.5Hz,1H),7.81(d,J=7. 6Hz,1H),7.71(ddd,J=12.5,9.6,4.1Hz,2H),7.60–7.54(m,1H),5.12(s,2H),3.30(q,J=5.8Hz,2H),2.97–2.71(m,4H),1.61–1.60(m,2H).

[0176] Synthesis of A25: 46 mg of compound 13-1 (0.22 mmol, 1.1 eq) and 90 mg of HATU (0.25 mmol, 1.25 eq) were added to a 10 mL two-necked flask. 4 mL of DCM was added and stirred to dissolve the compound. 300 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 87 mg of compound 26-2 (0.2 mmol, 1 eq) was added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until completion. The mixture was purified by column chromatography (DCM:MeOH = 130:1) to give 26 mg of a white solid, yield 22.1%. mp 135.0–135.8 °C. MS (ESI) + m / z measured value: 590.1281, calculated value: 590.1285 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.27–8.13(m,2H),7.96(d,J=8.8Hz,2H),7.92(d,J=15.0H z,1H),7.84(d,J=7.8Hz,1H),7.71(s,1H),7.57(t,J=7.3Hz,1H),7.50–7.45(m, 2H),7.33(t,J=5.1Hz,1H),7.15(d,J=5.4Hz,1H),7.01(d,J=15.0Hz,1H),6.91( d,J=8.8Hz,2H),4.81(s,2H),3.85(s,3H),3.53–3.42(m,4H),2.17–1.95(m,4H).

[0177] Example 27

[0178]

[0179] Synthesis of A26: 45 mg of cinnamic acid (0.3 mmol, 1.5 eq) and 152 mg of HATU (0.4 mmol, 2 eq) were added to a 10 mL two-necked flask. 4 mL of DCM was added and stirred to dissolve the mixture. 60 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 80 mg of compound 21-3 (0.2 mmol, 1 eq) was added, and the mixture was stirred for 4 h. The reaction was monitored by TLC until completion. The mixture was purified by column chromatography (DCM:MeOH = 80:1) to give 75 mg of a white solid, yield 70.8%. mp 129.3–129.7 °C. MS (ESI) + m / z measured value: 530.1075, calculated value: 530.1074 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.09(s,1H),8.01(s,1H),7.89(d,J=7.5Hz,1H),7.69(d,J=15.5Hz,1H),7.62(t,J=7.2 Hz,1H),7.51(t,J=7.5Hz,4H),7.36(d,J=6.0Hz,4H),6.87(d,J=12.5Hz,1H),4.84(s,2H),3.80-3.58(m,8H).

[0180] Example 28

[0181]

[0182] Synthesis of A27: 66 mg of compound 21-1 (0.2 mmol, 1 eq) and 83 mg of HATU (0.22 mmol, 1.1 eq) were added to a 10 mL two-necked flask. 4 mL of LCM was added and stirred to dissolve. 100 μL of DIPEA was added dropwise, and the mixture was stirred for 15 min. 45 mg of N-cinnamylpiperazine (0.22 mmol, 1.1 eq) was added, and the mixture was stirred for 4 h. The reaction was monitored by LC-MS until complete. The mixture was diluted with 3 mL of LCM, washed three times with 5 mL of saturated sodium bicarbonate solution, extracted, and the organic phase was washed three times with 5 mL of 1 M hydrochloric acid. The organic phase was dried over anhydrous Na₂SO₄ and evaporated to dryness. No further purification was required to obtain the product, 82 mg of white solid, yield 79.6%. mp 240.1–240.6 °C. MS (ESI) + m / z measured value: 516.1278, calculated value: 516.1281 [M+H] + . 1HNMR(500MHz,DMSO-d6)δ8.31(t,J=2.0Hz,1H),8.03–7.95(m,1H),7.81(d,J=7.5Hz,1H),7.7 4–7.70(m,1H),7.67(d,J=7.5Hz,1H),7.56(t,J=7.5Hz,1H),7.50(d,J=7.5Hz,2H),7.48–7.45 (m,1H),7.39(t,J=7.5Hz,2H),7.33(t,J=7.5Hz,1H),6.85(d,J=16.0Hz,1H),6.50–6.38(m,1H ),5.08(s,2H),3.92(d,J=6.0Hz,2H),3.67–3.41(m,4H),3.15(d,J=10.0Hz,2H),2.68(s,2H).

[0183] Example 29

[0184] Referring to Example 27, cinnamic acid was replaced with trifluoroacetic anhydride.

[0185]

[0186] Synthesis of A28: 50 mg of compound 21-3 (0.125 mmol, 1 eq) was added to a 5 mL test tube, and 1 mL of DCM was added and stirred to dissolve. 26 μL of triethylamine was added to the system, and the mixture was stirred for 10 min. 26 mg of trifluoroacetic anhydride was added to the system, and the mixture was stirred overnight. The reaction was monitored by TLC until completion. 3 mL of DCM was added for dilution, and the mixture was washed three times with 5 mL of water. The organic phase was extracted and separated. The organic phase was evaporated to dryness, redissolved in 5 mL of diethyl ether, filtered, and the filter cake was washed three times with 1 mL of diethyl ether. The filtrate was dried over anhydrous Na2SO4 to give 33 mg of a white solid, yield 53.3%. mp 134.4-134.7℃. MS (ESI) + m / z measured value: 496.0479, calculated value: 496.0478 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.18(s,1H),7.94(s,1H),7.91(d,J=7.9Hz,1H),7.64(dd,J=10.8 ,4.1Hz,1H),7.56–7.51(m,2H),7.37(d,J=1.3Hz,1H),4.86(s,2H),3.76-3.69(m,8H).

[0187] Example 30

[0188] Referring to Example 27, cinnamic acid was replaced with cyanoacetic acid.

[0189]

[0190] Synthesis of A29: 16 mg cyanoacetic acid (0.18 mmol, 1.2 eq) and 85 mg HATU (0.225 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 4 mL of LCM was added and stirred to dissolve the mixture. 54 μL of triethylamine was added dropwise, and the mixture was stirred for 15 min. 60 mg of compound 21-3 (0.15 mmol, 1 eq) was added to the mixture, and the mixture was stirred for 4 h. The reaction was monitored by TLC until complete. The mixture was diluted with 3 mL of LCM, washed three times with 5 mL of saturated NaHCO3, and the organic phase was extracted and separated. The organic phase was evaporated to dryness, redissolved in 2 mL of diethyl ether, and slurryed. The mixture was filtered, and the filter cake was washed three times with 1 mL of diethyl ether. The filter cake was dried to obtain 62 mg of a white solid, yield 88.7%. mp 237.4–238.2 °C. MS (ESI) + m / z measured value: 467.0713, calculated value: 467.0714 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ8.28(s,1H),7.95(s,1H),7.81(d,J=7.5Hz,1H),7.71(t,J=7.3Hz,1H),7.67(d,J =7.4Hz,1H),7.56(t,J=7.3Hz,1H),7.43(s,1H),5.08(s,2H),4.09(d,J=23.9Hz,2H),3.72–3.36(m,8H).

[0191] Example 31

[0192] In Example 27, cinnamic acid was replaced with chloroacetyl chloride.

[0193]

[0194] Synthesis of A30: 50 mg of compound 21-3 (0.125 mmol, 1 eq) was added to a 5 mL test tube, and 1 mL of DCM was added and stirred to dissolve. 26 μL of triethylamine was added to the system, and the mixture was stirred for 10 min. 21 mg of chloroacetyl chloride (0.188 mmol, 1.5 eq) was dissolved in 1 mL of DCM and added dropwise to the system. The reaction was stirred overnight, and the reaction was monitored by TLC until completion. The system was diluted with 3 mL of DCM, washed three times with 5 mL of saturated NH4Cl, and the organic phase was extracted and separated. The organic phase was evaporated to dryness, redissolved in 2 mL of diethyl ether, slurryed, filtered, and the filter cake was washed three times with 1 mL of diethyl ether. The filter cake was dried to give 50 mg of white solid, yield 84.2%. mp 125.5-125.7℃. MS (ESI) +m / z measured value: 476.0371, calculated value: 476.0371 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.13(d,J=13.3Hz,1H),7.97(s,1H),7.90(d,J=7.5Hz,1H),7.63(t,J= 7.3Hz,1H),7.52(t,J=7.0Hz,2H),7.36(s,1H),4.86(s,2H),4.09(s,2H),3.83-3.62(m,8H).

[0195] Example 32

[0196]

[0197] Synthesis of 32-1: 466 mg of N-Boc piperazine (2.5 mmol, 1 eq) was added to a 50 mL three-necked flask, purged three times with N2, followed by the addition of 10 mL anhydrous DCM and 400 μL triethylamine. The system was then placed in an ice bath. 571 mg of 2-chloroethylsulfonyl chloride (3.5 mmol, 1.4 eq) was dissolved in 10 mL anhydrous DCM and added dropwise to the system. The mixture was stirred for 30 min, then transferred to room temperature and stirred overnight. The reaction was monitored by LC-MS until completion. The system was evaporated to dryness at low temperature. The remaining oily liquid was added dropwise to 10 mL of 1 M sodium hydroxide aqueous solution, resulting in the precipitation of a large amount of white solid. The solid was filtered to give 389 mg of white solid, with a yield of 56.3%. 1 H NMR(500MHz,DMSO-d6)δ6.81(dd,J=16.5,10.0Hz,1H),6.19(d,J=10.0Hz,1H ),6.12(d,J=16.5Hz,1H),3.43–3.38(m,4H),3.03–2.96(m,4H),1.40(s,9H).

[0198] Synthesis of 32-2: 380 mmol of compound 32-1 was added to a 10 mL two-necked flask, followed by 0.675 mL of tetrahydrofuran with stirring to dissolve. 0.6 mL of a 4M solution of 1,4-dioxane hydrogen chloride was added dropwise to the system, and the reaction was stirred for 2 h. The reaction was monitored by LC-MS until completion. The mixture was filtered, and the filter cake was washed three times with 2 mL of tetrahydrofuran and dried to give 200 mg of a white solid, yield 67.0%. MS (ESI) + m / z measured value: 177.0694, calculated value: 177.0692 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ9.53 (s, 2H), 6.92 (dd, J=16.5, 10.0Hz, 1H), 6.26 (d, J= 10.0Hz,1H),6.17(d,J=16.5Hz,1H),3.32–3.28(m,4H),3.17(t,J=5.0Hz,4H).

[0199] Synthesis of A31: 79 mg of compound 21-1 (0.24 mmol, 1 eq) and 114 mg of HATU (0.3 mmol, 1.5 eq) were added to a 10 mL two-necked flask. 4 mL of LDCM was added and stirred to dissolve. 200 μL of LDIPEA was added dropwise, and the mixture was stirred for 15 min. 42 mg of compound 32-2 (0.2 mmol, 1.1 eq) was added to the mixture, and the mixture was stirred for 4 h. The reaction was monitored by LC-MS until complete. The mixture was evaporated to dryness, cooled, and recrystallized from the hot saturated methanol solution to give 45 mg of a white solid, yield 46.0%. mp 177.0–177.6 °C. MS (ESI) + m / z measured value: 490.0421, calculated value: 490.0421 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.15–8.09(m,1H),7.96(t,J=1.9Hz,1H),7.90(d,J=8.0Hz,1H),7.64(td,J=7.6,1.0Hz,1H),7.56–7.51(m,2H),7.33(t, J=1.4Hz,1H),6.45(dd,J=16.6,9.9Hz,1H),6.29(d,J=16.6Hz,1H),6.11(d,J=9.9Hz,1H),4.85(s,2H),3.86(s,2H),3.62(s,2H),3.23(s,4H).

[0200] Example 33

[0201]

[0202] Synthesis of 33-1: 851 mg cyanoacetic acid (10 mmol, 1 eq) was added to a 50 mL three-necked flask, followed by 10 mL of anhydrous pyridine and stirring to dissolve. 771 mg cyclopropaneformaldehyde (11 mmol, 1.1 eq) and 171 mg tetrahydropyrrole (2.4 mmol, 0.24 eq) were added to the system, and the mixture was stirred for 4 h. The reaction was monitored by LC-MS to indicate completion. The system was diluted with 30 mL of LEtOAc, washed three times with 10 mL of 6 M hydrochloric acid, extracted, and the organic phase was collected. The organic phase was dried over anhydrous Na₂SO₄ and evaporated to dryness. No further treatment was required to obtain 1.3 g of the product, with a yield of 94.9%. MS (ESI) + m / z measured value: 138.0548, calculated value: 138.0550 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ13.48(s,1H),7.17(d,J=11.4Hz,1H),1.97–1.89(m,1H),1.30–1.25(m,2H),1.06(dq,J=6.9,3.6Hz,2H).

[0203] Synthesis of A32: 25 mg of compound 33-1 (0.18 mmol, 1.2 eq) and 85 mg of HATU (0.225 mmol, 1.5 eq) were added to a 5 mL test tube, and 2 mL of DCM was added and stirred to dissolve. 45 μL of triethylamine was added to the system, and the mixture was stirred for 10 min. 60 mg of compound 21-3 was added to the system, and the reaction was stirred overnight. The reaction was monitored by LC-MS until complete. The system was evaporated to dryness, and the crude product was purified by column chromatography (DCM:MeOH = 130:1) to give 33 mg of a white solid, yield 42.5%. mp > 250 °C. MS (ESI) + m / z measured value: 519.1023, calculated value: 519.1027 [M+H] + . 1 H NMR(500MHz, CDCl3)δ8.06(dd,J=6.7,3.4Hz,2H),7.90(d,J=7.5Hz,1H),7.64(t,J=7.4Hz,1H),7.58–7.5 2(m,2H),7.41–7.32(m,2H),4.88(d,J=2.1Hz,2H),3.93–3.53(m,8H),1.35(s,4H),1.27(d,J=4.2Hz,1H).

[0204] Example 34

[0205]

[0206] Synthesis of 34-1: 46 mg of compound 21-3 (0.115 mmol, 1 eq) and 25 mg of N-Boc-4-piperidinone (0.121 mmol, 1.05 eq) were added to a 5 mL test tube, and 1 mL of DCE was added and stirred to dissolve. 7 μL of LAcOH (0.115 mmol, 1 eq) was added dropwise to the system, and the reaction was stirred for 1.5 h. Then, 22 mg of sodium cyanoborohydride (0.346 mmol, 3 eq) was added in portions, and the reaction was allowed to proceed for 24 h. The reaction was monitored by TLC until completion. The system was diluted with 10 mL of water and 10 mL of LEtOAC, and the organic phase was extracted and separated. The crude product was evaporated to dryness and purified by column chromatography (DCM:MeOH = 100:1) to give 20 mg of white solid, yield 85.9% MS (ESI). + m / z measured value: 583.1911, calculated value: 583.1915 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ9.09(s,1H),8.85(s,1H),8.30(s,1H),8.00(s,1H),7 .82(d,J=7.6Hz,1H),7.74–7.67(m,2H),7.57(t,J=7.3Hz,1H),7.50(s,1H),5. 09(s,2H),4.58(s,1H),3.76(s,2H),3.76-3.44(m,4H),3.17(s,2H),2.91(d, J=11.3Hz,2H),2.28(s,2H),1.96(d,J=12.7Hz,2H),1.40(s,1H),1.35(s,1H).

[0207] Synthesis of 34-2: 20 mg of compound 34-1 was added to a 10 mL two-necked flask, followed by 0.2 mL of tetrahydrofuran. The mixture was stirred to dissolve the compound. Then, 0.2 mL of a 4M solution of 1,4-dioxane hydrogen chloride was added dropwise. The mixture was stirred for 2 h, and the reaction was monitored by LC-MS until completion. The mixture was filtered, and the filter cake was washed three times with 2 mL of tetrahydrofuran and dried to give 17 mg of a white solid, with a yield of 95.5%. MS (ESI) + m / z measured value: 483.1888, calculated value: 483.1890 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ9.09(s,1H),8.85(s,1H),8.30(s,1H),8.00(s,1H),7 .82(d,J=7.6Hz,1H),7.74–7.67(m,2H),7.57(t,J=7.3Hz,1H),7.50(s,1H),5. 09(s,2H),4.58(s,1H),3.76(s,2H),3.76-3.44(m,4H),3.17(s,2H),2.91(d, J=11.3Hz,2H),2.28(s,2H),1.96(d,J=12.7Hz,2H),1.40(s,1H),1.35(s,1H).

[0208] Synthesis of A33: 17 mg of compound 34-2 (0.0328 mmol, 1 eq) was added to a 10 mL two-necked flask, followed by 2 mL of DCM and stirring until homogeneous. Then, 14 μL of triethylamine (0.82 mmol, 2.5 eq) was slowly added dropwise, and the mixture was stirred for 15 min. 7 mg of acryloyl chloride (0.656 mmol, 2 eq) was added dropwise, and the mixture was stirred for 2 h. The reaction was monitored by LC-MS until completion. The mixture was quenched with 5 mL of water, diluted with 5 mL of DCM, and washed three times with 5 mL of 1 M hydrochloric acid. The aqueous phase was collected. The aqueous phase was neutralized to pH 8 with saturated NaHCO3 solution, and extracted with 5 mL of EtOAc. The organic phase was collected by rotary evaporation. The product, a white solid, was obtained in 15 mg of 85.3% yield without further purification. The reaction temperature was 116.0–116.7 °C. MS(ESI+): Measured m / z value: 537.1499, Calculated value: 537.1496[M+H]+. 1H NMR (500MHz, CDCl3) δ8.07(t,J=1.9Hz,1H),7.98–7.95(m,1H),7.91(d,J=8.0Hz,1H),7.63(td,J=7.5,1.0Hz,1H) ,7.54–7.51(m,2H),7.33(t,J=1.5Hz,1H),6.57(dd,J=16.8,10.6Hz,1H),6.25(dd,J=16.8,1.9Hz,1H),5.67(dd, J=10.6,1.9Hz,1H),4.85(s,2H),4.68(d,J=12.4Hz,1H),4.04(d,J=13.1Hz,1H),3.78(s,2H),3.49(s,2H),3.06( t,J=12.5Hz,1H),2.65–2.54(m,4H),1.88(d,J=12.2Hz,2H),1.46(dd,J=21.4,11.5Hz,2H),1.25(t,J=7.1Hz,2H).

[0209] Example 35

[0210]

[0211] Synthesis of 35-1: 300 mg of compound 11-1 (0.783 mmol, 1 eq), 197 mg of 4-methylpyridine-3-boronate pinacol ester (0.901 mmol, 1.15 eq), 383 mg of cesium carbonate (1.175 mmol, 1.5 eq), and 114 mg (0.157 mmol, 0.2 eq) of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged three times with N2. A mixed solvent of 16 mL of 1,4-dioxane and 4 mL of water was added to the system. After stirring thoroughly, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was complete as monitored by LC-MS, the system was evaporated to dryness. The solution was diluted with 20 mL of LCM and 20 mL of water, extracted, and the organic phase was collected. The organic phase was dried over anhydrous Na2SO4, and the crude product was evaporated to dryness. No further purification was required before use in the next step. MS (ESI) + m / z measured value: 397.2235, calculated value: 397.2234 [M+H] + .

[0212] Synthesis of 35-2: 0.783 mmol of compound 35-1 was added to a 10 mL two-necked flask, and 3.5 mL of dichloromethane was added and stirred to dissolve. 0.783 mL of trifluoroacetic acid was added to the system, and the mixture was stirred overnight. The reaction was monitored by TLC until completion. The system was diluted with 5 mL of DCM, and 10 mL of water was added for extraction and separation of the aqueous phase. 1 M NaOH solution was added dropwise to the aqueous phase until pH 12 was reached. 10 mL of DCM was added for extraction and separation of the organic phase. The organic phase was dried over anhydrous Na₂SO₄ and evaporated to dryness. No further purification was required to obtain 170 mg of the product, a reddish-brown solid, with a yield of 78.3%. MS (ESI) + m / z measured value: 297.1721, calculated value: 297.1720 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.43(d,J=5.0Hz,1H),8.39(s,1H),7.16(d,J=5.0Hz,1H),6.73(dd,J=2.0,1.5Hz,1H),6.65( t,J=1.5Hz,1H),6.64–6.62(m,1H),3.89(s,2H),3.73(s,2H),3.45(s,2H),2.92(s,2H),2.80(s,2H),2.28(s,3H).

[0213] Synthesis of A34: 49 mg of compound 13-1 (0.238 mmol, 0.95 eq) and 114 mg of HBTU (0.3 mmol, 1.2 eq) were added to a 10 mL two-necked flask. 2.5 mL of DCM was added and stirred to dissolve. 75 μL of triethylamine was added dropwise, and the mixture was stirred for 15 min. 74 mg of compound 35-2 (0.25 mmol, 1 eq) was added to the mixture, and the mixture was stirred for 3 h. The reaction was monitored by LC-MS until complete. The mixture was diluted with 5 mL of DCM, washed three times with 5 mL of saturated NH4Cl solution, extracted, and the organic phase was collected. The liquid was evaporated to dryness, and the crude product was purified by column chromatography using DCM:MeOH = 50:1 to give 63 mg of a white-yellow solid, yield 52.0%. mp 139.6-139.9℃. MS (ESI) + m / z measured value: 485.2186, calculated value: 485.2184 [M+H] + . 1H NMR (500MHz, CDCl3) δ8.43(d,J=4.9Hz,1H),8.38(d,J=11.4Hz,1H),8.16–7.82(m,3H),7.60–7.32(m,1H),7.18(d,J=4.8Hz, 1H),6.97(d,J=8.7Hz,2H),6.79–6.71(m,1H),6.70–6.61(m,2H),3.88(s,3H),3.81–3.52(m,8H),2.28(s,3H),1.88(s,2H).

[0214] Example 36

[0215]

[0216] Synthesis of A35: 48 mg of A34 (0.1 mmol, 1 eq) was added to a 10 mL two-necked flask, and 1 mL of LCM was added and stirred to dissolve. 8 μL of acetyl chloride (0.11 mmol, 1.1 eq) was dissolved in 1 mL of LCM and added dropwise to the system. The reaction was stirred for 2 h, and the reaction was monitored by TLC until completion. The system was diluted with 5 mL of LCM, quenched with 5 mL of water, neutralized with 1 M hydrochloric acid, and the organic phase was extracted and separated. The solution was evaporated to dryness, and 33 mg of a white-yellow solid was obtained without further purification, yielding 62.7% (mp 134.9-135.1℃). MS (ESI) + m / z measured value: 527.2285, calculated value: 527.2289 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.47(d,J=5.1Hz,1H),8.41(s,1H),8.04(d,J=8.8Hz,2H),7.98(d,J=14.9Hz,1H),7.87–7.76(m,2H),7.58– 7.42(m,2H),7.25(d,J=5.4Hz,1H),7.13(s,1H),6.98(d,J=8.9Hz,2H),3.89(s,3H),3.85–3.49(m,8H),2.32(s,3H),2.22(s,3H).

[0217] Example 37

[0218]

[0219] Synthesis of 37-1: 1.501 g of p-methoxyacetophenone (10 mmol, 1 eq) and 2.219 g of selenium dioxide (20 mmol, 2 eq) were added to a 25 mL three-necked flask. The mixture was purged with N2 three times, followed by the addition of 5 mL of pyridine. The mixture was stirred overnight, and the reaction was monitored by TLC until completion. The system was diluted with 15 mL of 1M LetOAc, washed once with 15 mL of 1M NaOH solution, and the aqueous phase was extracted and separated. The aqueous phase was adjusted to pH 1 with 1M hydrochloric acid, washed three times with 15 mL of 1M LetOAc, extracted, and the organic phases were combined and evaporated to dryness. No further purification was required to obtain 1.756 g of the product, an off-white solid, in 97.6% yield. 1 H NMR (500MHz, DMSO-d6) δ7.91 (d, J = 8.9 Hz, 2H), 7.14 (d, J = 8.9 Hz, 2H), 3.88 (s, 3H).

[0220] Synthesis of A36: 270 mg of compound 37-1 (1.5 mmol, 1 eq) was added to a 50 mL round-bottom flask, followed by 3 mL of anhydrous DCM and stirring to dissolve. Under ice bath conditions, 286 mg of oxaloyl chloride (2.25 mmol, 1.5 eq) and one drop of anhydrous DMF were added dropwise, and the mixture was stirred for 15 min. The mixture was slowly brought to room temperature and stirred for 2 h. The reaction was monitored by LC-MS until complete, and the mixture was directly rotary evaporated until the residual liquid no longer decreased. The mixture was then dissolved again in 5 mL of anhydrous DCM, and 260 μL of triethylamine (1.8 mmol, 1.2 eq) was added dropwise, followed by stirring for 10 min. 296 mg of compound 35-2 (1 mmol, 0.67 eq) was added fractionally, and the mixture was stirred for 2 h. The reaction was monitored by LC-MS until complete. The mixture was evaporated to dryness, and purified by column chromatography (DCM:MeOH = 50:1). The purified solid was dried to give 140 mg of a white-yellow solid, yield 45.6%. mp 101.4–101.6 °C. MS(ESI + m / z measured value: 459.2019, calculated value: 459.2027 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.40(d,J=26.7Hz,2H),7.90(d,J=8.3Hz,2H),7.19(d,J=3.1Hz,1H),6.97(d, J=8.4Hz,2H),6.72(s,1H),6.64(s,2H),4.16(s,2H),3.88(s,3H),3.82–3.32(m,8H),2.27(s,3H).

[0221] Example 38

[0222]

[0223] Synthesis of 38-1: 322 mg of 3-amino-5-bromobenzoic acid (1.5 mmol, 1 eq), 378 mg of 4-methylpyridine-3-boronic acid pinacol ester (1.725 mmol, 1.15 eq), 733 mg of cesium carbonate (2.25 mmol, 1.5 eq), and 218 mg (0.3 mmol, 0.2 eq) of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride were added to a 25 mL three-necked flask. The mixture was purged three times with N2. A mixed solvent of 16 mL of 1,4-dioxane and 4 mL of water was added to the system. After stirring thoroughly, the mixture was slowly heated to 65 °C and reacted for 1 h. After the reaction was complete as monitored by LC-MS, the system was evaporated to dryness. The solution was diluted with 20 mL of LDC and 20 mL of water, extracted, and the aqueous phase was collected and evaporated to dryness. The residue was redissolved in 20 mL of LETOAc, filtered, and the organic phase was collected and evaporated to dryness. No further purification was required for the next step. MS (ESI) + m / z measured value: 229.0973, calculated value: 229.0972 [M+H] + .

[0224] Synthesis of 38-2: 360 mg of compound 37-1 (2 mmol, 1 eq) was added to a 50 mL round-bottom flask, and 4 mL of anhydrous DCM was added and stirred to dissolve. Under ice bath conditions, 256 μL of oxaloyl chloride (3 mmol, 1.5 eq) and one drop of anhydrous DMF were added dropwise, and the mixture was stirred for 15 min. The mixture was slowly brought to room temperature and stirred for 2 h. The reaction was monitored by LC-MS until complete, and the mixture was directly rotary evaporated until the residual liquid no longer decreased. The mixture was then dissolved again in 5 mL of anhydrous DCM, and 346 μL of triethylamine (2.4 mmol, 1.2 eq) was added dropwise, and the mixture was stirred for 10 min. 478 mg of N-Boc piperidinylmethylamine (2.2 mmol, 1.1 eq) was added in portions, and the mixture was stirred for 2 h. The reaction was monitored by LC-MS until complete. The system was diluted with 5 mL of DCM, washed three times with 5 mL of 1 M hydrochloric acid, extracted, and the organic phase was separated, dried over anhydrous Na₂SO₄, and evaporated to dryness. No further purification was required to obtain 540 mg of the product as a white-yellow solid, with a yield of 71.8%. MS (ESI) + m / z measured value: 775.3886, calculated value: 775.3888 [2M+Na] + . 1 H NMR (500MHz, CDCl3) δ8.58–8.27(m,2H),6.96–6.93(m,2H),4.11(dd,J=14.0,7.0Hz,2H),3.89 (s,3H),3.28(t,J=6.0Hz,2H),2.69(t,J=10.5Hz,2H),1.81–1.66(m,4H),1.47–1.42(m,11H).

[0225] Synthesis of 38-3: 489 mg of compound 38-2 (1.404 mmol, 1 eq) was added to a 50 mL two-necked flask, followed by 3.5 mL of tetrahydrofuran and stirring to dissolve. Then, 3.5 mL of a 4M solution of 1,4-dioxane hydrogen chloride was added dropwise to the system, and the reaction was stirred for 2 h. The reaction was monitored by LC-MS until completion. The mixture was filtered, and the filter cake was washed three times with 2 mL of tetrahydrofuran and dried to obtain 322 mg of white solid. No further purification was required before proceeding to the next step, with a yield of 83.1%. MS (ESI) + m / z measured value: 277.1544, calculated value: 277.1547 [M+H] + .

[0226] Synthesis of A37: 91 mg of compound 38-1 (0.4 mmol, 1 eq), 131 mg of compound 38-3 (0.46 mmol, 1.15 eq), and 182 mg of HBTU (0.48 mmol, 1.2 eq) were added to a 25 mL three-necked flask. 5 mL of DCM was added and stirred to dissolve the mixture. 144 μL of triethylamine was added dropwise to the system, and the reaction was stirred overnight. The reaction was monitored by LC-MS until complete. The system was evaporated to dryness, and the crude product was purified by column chromatography (DCM:MeOH = 30:1) to give 100 mg of white solid, yield 54.6%. mp 154.0–155.0 °C. MS (ESI) + m / z measured value: 487.2339, calculated value: 487.2340 [M+H] + . 1 H NMR (500MHz, CDCl3) δ8.43 (s, 1H), 8.40 (d, J = 9.0Hz, 2H), 7.30 (t, J = 6.0Hz, 1H) ,7.19(d,J=4.6Hz,1H),6.94(d,J=9.0Hz,2H),6.72(d,J=1.3Hz,1H),6.66–6.61 (m,2H),4.71(s,1H),3.87(d,J=14.1Hz,5H),3.35–3.28(m,2H),3.03–2.92(m, 2H),2.82–2.72(m,1H),2.29(s,3H),1.92–1.84(m,2H),1.25(t,J=14.6Hz,4H).

[0227] Example 39 Affinity of the compound provided by the present invention to MYC protein

[0228] All compounds involved in this invention were subjected to 16 1 / 2-dilute gradients from 100 μM using PBS-T buffer. After dilution, 200 nM GFP-N-Myc-His or GFP-C-Myc-His recombinant protein was mixed with the compounds at a 1:1 volume ratio (10 μL + 10 μL in this experiment), thoroughly mixed, and allowed to stand for 5 min. Capillary sampling was used to simultaneously detect MST in 16 groups using a Nanotemper NT 11.5 instrument. Data were analyzed using MO.Analysis software at 20-second intervals, and the curve fitting model was the Kd Model. The final K... d The values ​​were calculated and exported by the software. See Tables 1 and 2 below for specific results. Figure 1 .

[0229] Table 1. K-related properties of the compounds provided in this invention with N-Myc d value

[0230]

[0231]

[0232] Table 2. K-type of the compound provided in this invention and C-Myc d value

[0233] Compound numbering <![CDATA[K d / M]]> Compound numbering <![CDATA[K d / M]]> A00 <![CDATA[1.72±0.89×10 -7 ]]> A10 <![CDATA[5.39±2.10×10 -8 ]]> A09 <![CDATA[2.39±1.02×10 -7 ]]> A12 <![CDATA[2.14±0.61×10 -7 ]]>

[0234] Based on the results in Tables 1 and 2, the compounds provided by this invention exhibit good affinity for N-Myc, K d The value is 1.25 ± 0.74 × 10 -5 ~4.61±1.68×10 -8 Some compounds also exhibit good binding affinity with C-Myc, K d The value is 1.72 ± 0.89 × 10 -7 ~5.39±2.10×10 -8 .

[0235] Example 40: Inhibitory activity of the compounds provided in this invention against the proliferation of NMYC-amplified tumor cells.

[0236] MYCN-amplified tumor cell lines SK-N-BE(2) and RH30 were seeded in 96-well plates. The drug was diluted 10 μM at six different concentrations, and administered at a cell density of 50%. After 72 hours of drug treatment, CCK8 reagent was added at a 1:10 ratio, and the reaction was carried out at 37°C for approximately 2-3 hours. The absorbance of each group was then measured at 450 nm using a microplate reader. The cell proliferation inhibition at different concentrations was calculated, and IC50 curves were plotted to calculate the IC50 value. 50The specific results are shown in Table 3 below.

[0237] Table 3 shows the IC50 values ​​of the compounds provided in this invention on the SK-N-BE(2) cell line. 50

[0238] Compound numbering <![CDATA[IC 50 (μM)]]> Compound numbering <![CDATA[IC 50 (μM)]]> A11 6.76 A22 0.27 A12 0.46±0.15 A23 0.18 A16 7.58 A24 8.05 A18 4.02 A25 6.01 A19 0.38 A30 2.13 A20 0.38 A34 1.06±0.28 A21 0.46 A35 0.20

[0239] Table 4. IC50 of the compounds provided in this invention on the RH30 cell line. 50

[0240] Compound numbering <![CDATA[IC 50 (μM)]]> A12 0.48 A34 0.28

[0241] Example 41: Degradation activity of the compounds provided in this invention against NMYC protein.

[0242] SK-N-BE(2) cell lines were seeded in 24-well plates and treated with the compound provided in this invention at 50% cell density using a gradient concentration for a uniform treatment time of 12 h. GAPDH was used as an internal control. Cell samples were collected at 1.5× Loading and denatured in a 95°C metal bath. Changes in N-Myc protein levels in cells after compound treatment were detected by Western blot. Specific results are attached below. Figure 2 As shown, most of the compounds can cause the degradation of intracellular NMYC protein, among which A12, A19, A21, A22, A25, A34 and A35 can reduce the content of intracellular NMYC protein in a dose-dependent manner.

[0243] Example 42 Pharmacokinetic study of the compound provided in this invention in rats

[0244] Nine SD rats were randomly divided into three groups and administered compound A12 via intravenous injection (iv), intraperitoneal injection (ip), or oral administration (po) at a dose of 5 mg / kg, respectively. Blood samples (approximately 0.25 mL) were collected via tail vein at the following time points: 5 min, 10 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after intravenous administration. Blood samples were collected at the same time points for the intraperitoneal injection and oral administration groups. The concentration of compound A12 in rat plasma samples was determined using LC-MS / MS, and pharmacokinetic parameters were calculated using Phoenix WinNolin software. Specific results are shown in Table 5 below, including the time to peak concentration (ts) of A12 in rats. max The shorter (h) indicates that the A12 compound has a faster onset of action and appropriate bioavailability after oral and intraperitoneal administration.

[0245] Table 5. Pharmacokinetic parameters of compound A12 provided by this invention

[0246]

[0247] Example 43: Antitumor activity of the compounds provided in this invention in a mouse model of xenograft tumors.

[0248] Approximately 5 million RH30 tumor cells were collected and inoculated into the axilla (unilaterally) of nude mice. The tumors were allowed to grow to a size of approximately 50-100 mm. 3 Compound A12 was administered intraperitoneally starting at a certain time, and tumor volume in mice was measured and recorded daily. The tumor volume was approximately 1000 mm². 3 As the experimental endpoint, mice were euthanized, dissected, and tumors were removed, photographed, and weighed. Vincristine, a first-line clinical chemotherapy drug, was selected as the positive control, and the animal dosage was converted from the clinical dosage. The relative tumor volume (RTV) was calculated based on the measurements using the formula: RTV = Vt / V0. Where V0 is the tumor volume measured at the time of administration (d0), and Vt is the tumor volume at each measurement. The evaluation index of antitumor activity was the relative tumor proliferation rate (T / C%), calculated using the formula: T / C = T RTV / C RTV ×100% (T) RTV Treatment group RTV; C RTV (Negative control group RTV)

[0249] The specific results are shown in Table 6 below. Figure 3 Compound A12 showed dose-dependent growth inhibition of RH30 xenograft tumors, and the growth inhibition rate of A12 in the high-dose group was slightly better than that of the vincristine group.

[0250] Table 6. Growth inhibitory effect of compound A12 on RH30 xenograft tumors

[0251]

[0252] Cut 1mm 2 -2mm 2 SK-N-DZ cell-derived xenograft tumor tissue blocks were injected twice into the axilla (unilaterally) of nude mice using a cannula. The tumors were allowed to grow to approximately 50-100 mm in size. 3 Compound A12 was administered intraperitoneally starting at a certain time, and tumor volume in mice was measured and recorded daily. The tumor volume was approximately 2000 mm². 3As the experimental endpoint, mice were euthanized, dissected, and the tumors were removed, photographed, and weighed. Cisplatin, a first-line clinical chemotherapy drug, was selected as a positive control. The animal dosage was converted from the clinical dosage, and the detection indicators were the same as above. The specific results are shown in Table 7 below. After administration of compound 12, the growth inhibition effect of SK-N-DZ xenograft tumors was dose-dependent, and the tumor inhibition rates of both the high and low doses of compound 12 were significantly better than those of positive cisplatin.

[0253] Table 7. Growth inhibitory effect of compound A12 on SK-N-DZ xenograft tumors

[0254]

[0255] Cut 1mm 2 -2mm 2 SK-N-BE(2) cell-derived xenograft tumor tissue blocks were inoculated twice into the axilla (unilaterally) of nude mice using a cannula. The tumors were allowed to grow to approximately 50-100 mm in size. 3 Compound A12 was administered intraperitoneally starting at a certain time, and tumor volume in mice was measured and recorded daily. The tumor volume was approximately 2000 mm². 3 As the experimental endpoint, mice were euthanized, dissected, and the tumors were removed, photographed, and weighed. Cisplatin, a first-line clinical chemotherapy drug, was selected as a positive control. The animal dosage was converted from the clinical dosage, and the detection indicators were the same as above. The specific results are shown in Table 8 below. After administration of compound 12, the growth of SK-N-DZ xenograft tumors was significantly inhibited, which was superior to that of positive control cisplatin.

[0256] Table 8. Growth inhibitory effect of compound A12 on SK-N-BE(2) xenograft tumors

[0257]

[0258] Cut 1mm 2 -2mm 2 Tumor xenografts from patients with recurrent, metastatic, or chemotherapy-resistant neuroblastoma were injected twice into the axilla (unilaterally) of nude mice using a cannula. Once the tumor had grown to approximately 50-100 mm in size... 3 Compound A12 was administered intraperitoneally starting at a certain time, and tumor volume in mice was measured and recorded daily. The tumor volume was approximately 1000 mm². 3 As the experimental endpoint, mice were euthanized, dissected, and the tumors were removed, photographed, and weighed. Cisplatin, a chemotherapy drug, was selected as a positive control. Animal dosages were converted from clinical dosages, and the detection indicators were the same as above. Specific results are shown in Table 9 below. After administration of compound 12, the growth inhibition effect on neuroblastoma patient-derived xenograft tumors was dose-dependent, and the tumor inhibition rates of both high and low doses of compound 12 were significantly better than those of cisplatin.

[0259] Table 9. Growth inhibitory effect of compound A12 on neuroblastoma patient-derived xenograft tumors

[0260]

Claims

1. A compound having the structure shown in general formula I or a pharmaceutically acceptable salt or isotope-labeled compound thereof: A is a substituted or unsubstituted acetamide group or a cyclized amide group or amino group; when substitution is present, the substituent is hydrogen, C1-C5 alkyl, C1-C5 heterocyclic alkyl, C6-C10 aryl, C1-C3 alkoxy or halogen; B is an unsubstituted, substituted acetamide subunit, or none at all; if substitution is present, the substituent is a C1-C5 alkyl, a C6-C10 aryl, or a C1-C3 alkoxy group. M is a carbon or nitrogen atom; C is hydrogen, halogen, 3-8 membered heterocyclic alkyl, C6-C10 aryl, 5-10 membered heteroaryl, or absent; the heterocyclic alkyl, aryl, and heteroaryl may be further substituted by C1-C3 alkyl or C1-C3 alkoxy groups. R1 and R2 are each independently hydrogen, C1 to C5 alkyl, or R1 and R2 together with the N atom attached to them to form a 5-7 membered ring; the alkyl group may be further substituted with a 3- to 8-membered heterocyclic alkyl group or an amino group; the 5-7 membered ring may optionally be substituted with one or more substituents selected from C1 to C3 alkyl groups or C1 to C3 alkyl groups containing heteroatoms. D represents a structure that binds to E3 ubiquitin ligase or heat shock protein, or it may not exist.

2. The compound according to claim 1, characterized in that, A is Wherein, R3 is hydrogen, C1-C5 alkyl, C1-C5 heterocyclic alkyl, or C6-C10 aryl; E is C6-C10 aromatic ring, 5-6 membered heteroaromatic ring or absent; R4 is hydrogen, C1-C3 alkyl, C1-C3 alkoxy or halogen; n = 1-2; B is Or not present; wherein, R5 is a C1-C5 alkyl, C6-C10 aryl, or C1-C3 alkoxy; m = 1-5; *The corresponding C atom is an S configuration or an R configuration or a mixture of S and R configurations; C is hydrogen, halogen, 4-6 membered heterocyclic alkyl, C6-C10 aryl, 5-6 membered heteroaryl or absent; wherein, the C6-C10 aryl or 5-6 membered heteroaryl may be further substituted by one or more substituents selected from halogen, hydroxyl, C1-C3 alkyl or C1-C3 alkoxy. for Among them, R1 and R6 are each independently hydrogen and C1 to C5 alkyl groups; R7 and R8 are each independently -CH2-, -NH-, and -NR. 11 -, O, S; R9 is H, C1-C5 alkyl, and there can be two or more R9s; Q is -CH- or -N-; x = 1-5; y = 1-4; z = 1-4; R 11 It is an alkyl group of C1 to C3; D represents a small ligand that binds to CRBN, a small ligand that binds to VHL, a covalent fragment that binds to other E3 ubiquitin ligases, a small ligand that binds to HSP70, or is absent.

3. The compound according to claim 1 or 2, characterized in that, The compound has a structure shown in any one of general formulas I-1 to I-16 or a pharmaceutically acceptable salt or isotope-labeled compound thereof; R1、R3、R5、R6、R9、x、n、 Same as the definition in claim 1 or 2; x a x b x c x d x e Each group is independently selected from hydrogen, halogen, cyclopropyl, cyano, carbonyl, carbamoyl, or C6-C10 arcarbamoyl; wherein the arcarbamoyl group may be further substituted by one or more substituents selected from halogen, hydroxyl, C1-C3 alkyl, or C1-C3 alkoxy.

4. The compound according to claim 1, characterized in that, A is selected from or amino; B is selected from Or it may not exist; C is H or Br; D is H or Selected from -NHMe, 5. The compound according to claim 1, characterized in that, The compound has one of the following structures or a pharmaceutically acceptable salt or isotope-labeled compound thereof:

6. A pharmaceutical composition comprising a compound as claimed in any one of claims 1-5, or a pharmaceutically acceptable salt thereof, an isotopically labeled compound thereof, and at least one pharmaceutically acceptable carrier.

7. The compound of any one of claims 1-5 or a pharmaceutically acceptable salt thereof, an isotopically labeled compound, or the pharmaceutical composition of claim 6, for use in preparing a degrader targeting the MYC protein.

8. Use of the compound of any one of claims 1-5 or a pharmaceutically acceptable salt thereof, an isotopically labeled compound thereof, or the pharmaceutical composition of claim 6 in the preparation of a medicament for treating diseases associated with MYC gene amplification, fusion, mutation, expression imbalance, allosteric variation, and functional abnormalities.

9. The use as described in claim 8, wherein the disease is neuroblastoma or rhabdomyosarcoma.

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