Compound containing disubstituted 7-azaindole structure and application

Abstract

The invention discloses a compound containing a disubstituted 7-azaindole structure, which is a compound with a structure as shown in a formula (I) or a salt of the compound or a solvate of the compound. The compound # imgabs0 has an obvious inhibiting effect on ATM, so that the compound # imgabs0 can be used as an ATM inhibitor for preparing medicines for preventing and treating ATM-mediated diseases, such as colorectal cancer, and can obviously inhibit tumor growth. Moreover, the compound can also be combined with liposome irinotecan for use, so that the tumor growth inhibition rate can be further improved and reaches 95.4%.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly relates to a class of compounds containing a disubstituted 7-azaindole structure and their application in the preparation of ataxia-telangiectasia mutated kinase (ATM) inhibitors in DNA damage response. Background Art

[0002] Genomic instability is a potential feature of tumor cells, which is associated with the accumulation of DNA damage in tumor cells during excessive proliferation and differentiation. Research shows that there are mainly three differences in the DDR (DNA damage response) pathway between tumor cells and normal cells. (1) One or more DDR pathways are lost during the occurrence and development of most cancers, leading to a greater dependence of tumor cells on the remaining DDR pathways. Inhibition of the remaining pathways exhibits significant cytotoxicity on tumors with other DDR function defects, thus providing a "synthetic lethality" treatment strategy. (2) The slowdown or stalling of replication forks caused by the accumulation of DNA damage leads to the dependence of cancer cells on the replication stress repair pathway. By inhibiting the excessive replication stress response, cancer cells may enter mitosis with excessive DNA damage and ultimately die. (3) Multiple factors related to the excessive proliferation of tumor cells exacerbate endogenous DNA damage, and jointly drive tumor cells to experience high levels of replication stress and transcriptional function under the action of hormones and inflammation, resulting in genomic instability. The result of these differences means that cancer cells are more sensitive to exogenous DNA damage than normal cells, supporting the hypothesis that targeting the DDR pathway may provide an effective approach for cancer treatment.

[0003] ATM kinase (ataxia–telangiectasia mutated kinase) is a serine / threonine protein kinase of approximately 350 kDa and is a member of the highly conserved PIKK (phosphatidylinositol 3 kinase-related kinases) protein family, which is commonly found in a variety of cells and organisms. ATM is a key regulatory kinase in the DDR pathway, and its main function is to respond to DSB (DNA double-strand break) signals and participate in DNA repair (by HR or NHEJ), activation of cell cycle checkpoints, chromatin remodeling, cellular senescence, apoptosis, etc. ATM usually exists as a dimer in normal cells; after DNA DSB damage occurs, histone H2AX in chromatin is rapidly phosphorylated to form γH2AX, and the DNA end-joining MRN (MRE11-RAD50-NBS1) complex is recruited to the damage site. Subsequently, the HEAT domain at the N-terminus of the ATM protein recognizes and binds to the C-terminus of the NBS1 protein, and a stable MRN-ATM complex is formed through the interaction of arginine residues with the MR complex (MRE11-RAD50). After being recruited by MRN, ATM is rapidly activated through autophosphorylation at at least four residue sites (Ser367, Ser1893, Ser1981, and Ser2996) and acetylation by the acetyltransferase TIP60 at Lys3016, thereby promoting its monomerization and kinase activity. The phosphorylation of ATM can activate hundreds of downstream substrates and regulate cellular life activities through multiple signaling pathways.

[0004] Gene sequencing results show that the ATM gene is the most common abnormal gene in sporadic cancers. 722 ATM mutation sites were reported in 108 representative cancer genome studies, distributed in various domains of ATM. At the same time, ATM mutations have been found in various types of tumors. Among them, the mutation rate in mantle cell lymphoma (MCL) is approximately 40%, and the mutation rate in tumors such as cutaneous squamous cell carcinoma (CSCC), small cell carcinoma of the lung (SCLC), uterine tumors, and colorectal cancer is approximately 20%. The vast majority of these mutations are missense mutations. Therefore, ATM inhibitors are expected to be used in the treatment of various diseases such as colorectal cancer. Summary of the Invention

[0005] The object of the present invention is to solve the above technical problems and provide a class of compounds containing a disubstituted 7-azaindole structure and their applications.

[0006] The technical solution adopted in the present invention is as follows: A class of compounds containing a disubstituted 7-azaindole structure is a compound having the structure shown in formula (Ι) or a salt of the compound, or a solvate of the compound;

[0007]

[0008] Wherein, R1 is selected from one of hydrogen, halogen, amino, cyano, acyl, sulfonyl, ureido or substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkyl, five-membered or six-membered nitrogenous saturated alkyl;

[0009] Ring A is selected from one of a substituted or unsubstituted benzene ring, a substituted or unsubstituted nitrogen-containing heterocycle, and a substituted or unsubstituted aromatic fused ring.

[0010] Preferably, ring A is selected from one of them.

[0011] Preferably, the pyridine substitution in ring A is a 2,5-disubstitution, and the benzene ring is a para-substitution.

[0012] Preferably, the R1 is a substituted C1-C5 alkoxy or a six-membered nitrogenous saturated alkyl.

[0013] More preferably, the compound is one of the following structures:

[0014]

[0015] The present invention also provides the application of the above-mentioned compound containing a disubstituted 7-azaindole structure in the preparation of a drug for preventing or treating ATM-mediated diseases.

[0016] Preferably, the disease is colorectal cancer.

[0017] More preferably, the compound is used in combination with liposomal irinotecan.

[0018] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) naming system.

[0019] Regarding the definition of the terms used in the present invention: Unless otherwise specified, the initial definitions provided for the groups or terms herein apply to the groups or terms throughout the specification; for terms not specifically defined herein, meanings that can be given by those skilled in the art should be provided according to the disclosure and context.

[0020] "Substituted" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.

[0021] The minimum and maximum carbon atom contents in a hydrocarbon group are indicated by a prefix. For example, the prefix Ca-Cb alkyl indicates any alkyl group containing "a" to "b" carbon atoms. Thus, for example, "C1-C6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms; "C1-C6 alkoxy" refers to an alkoxy group containing 1 to 6 carbon atoms.

[0022] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of carbon atoms. For example, C1-C8 alkyl refers to an alkyl group having 1 to 8 carbon atoms, i.e., an alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. The alkyl group can be straight-chain or branched-chain. Representative branched-chain alkyl groups have one, two, or three branches. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl, etc.

[0023] "Alkenyl" refers to an aliphatic hydrocarbon group having at least one carbon-carbon double bond. All alkenyl groups can be straight-chain or branched-chain.

[0024] "Alkynyl" refers to an aliphatic hydrocarbon group having at least one carbon-carbon triple bond. All alkynyl groups can be straight-chain or branched-chain.

[0025] The structure of "alkoxy" is a1 is an integer from 0 to 5.

[0026] "Halogen" is fluorine, chlorine, bromine, or iodine.

[0027] "Heterocycloalkyl" means that at least one carbon atom on the cycloalkyl ring is replaced by a heteroatom, and the heteroatom is O, N, or S, such as including but not limited to:

[0028] The beneficial effects of the present invention: The present invention provides a class of compounds containing a disubstituted 7-azaindole structure. This compound has an obvious inhibitory effect on ATM. Therefore, it can be used as an ATM inhibitor for preparing drugs for preventing and treating ATM-mediated diseases, such as colorectal cancer, and can significantly inhibit tumor growth. Moreover, this compound can also be used in combination with liposomal irinotecan, which can further increase the tumor growth inhibition rate to 95.4%. Description of the Drawings

[0029] Figure 1 It is the in vitro anti-tumor activity of compound YT-11.

[0030] Figure 2Inhibition of the ATM signaling pathway by compound YT-11.

[0031] Figure 3 Antitumor activity of compound YT-11 in vivo. Detailed implementation mode

[0032] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0033] Example 1 Synthesis of Intermediates 1 and 2

[0034]

[0035] 1. Reagents and conditions

[0036] (a) PdCl2(dppf), K2CO3, Dioxane / H2O / EtOH, 85 °C, 4 h; (b) THF, NIS, rt, 3 h; (c) PdCl2(dppf), K2CO3, Dioxane / H2O / EtOH, 75 °C, 4 h.

[0037] 2. Synthesis of Intermediate 1

[0038] Add 5-bromo-7-azaindole (2 g, 10 mmol), 4-isopropylsulfonylphenylboronic acid pinacol ester (4 g, 13 mmol), potassium carbonate (3.5 g, 25 mmol) and bis(diphenylphosphino)ferrocene dichloropalladium (0.73 g, 1 mmol) to a 250 mL three-necked flask. Add a mixed solvent system composed of dioxane and water (110 mL, volume ratio 7:4), and then heat the mixture to 85 °C under a nitrogen atmosphere and maintain for 4 hours. Subsequently, cool the reaction mixture to room temperature, and the crude product is purified by flash column chromatography. After concentration, the desired product is obtained as a white solid with a yield of 78%. 1 H NMR (400 MHz, DMSO-d6) δ 11.34 (s, 1H), 8.82 (d, J = 2.1 Hz, 1H), 8.17 (d, J = 8.3 Hz, 2H), 8.09 (d, J = 2.1 Hz, 1H), 7.95 (d, J = 8.2 Hz, 2H), 7.85 (d, J = 2.5 Hz, 1H), 7.03 (dd, J = 7.5 1.4 Hz, 1H), 3.50 (p, J = 6.8 Hz, 1H), 1.25 (d, J = 6.8 Hz, 6H).

[0039] 3. Synthesis of Intermediate 2

[0040] Dissolve intermediate 1 (2.3 g, 7.8 mmol) in tetrahydrofuran (50 mL) in a 100-mL round-bottom flask. After stirring the mixture for 5 minutes, add N-iodosuccinimide (NIS) (2.3 g, 10 mmol) to the reaction vessel. The reaction mixture was maintained at room temperature for about 3 hours, and the reaction progress was monitored by thin-layer chromatography (TLC). After the reaction was completed, the mixture was concentrated under reduced pressure to obtain a dark yellow solid. Subsequently, the crude product was dissolved in saturated sodium thiosulfate solution (100 mL) and extracted with ethyl acetate (100 mL × 4). The combined organic phases were concentrated under reduced pressure, and the resulting solid was dried to obtain intermediate 2 (2.88 g). 1 HNMR (400 MHz, DMSO-d6) δ 12.30 (s, 1H), 8.65 (d, J = 2.1 Hz, 1H), 8.07 (d, J = 8.3 Hz, 2H), 7.98 (d, J = 2.1 Hz, 1H), 7.93 (d, J = 8.2 Hz, 2H), 7.81 (d, J = 2.5 Hz, 1H), 3.46 (p, J = 6.8 Hz, 1H), 1.19 (d, J = 6.8 Hz, 6H).

[0041] Synthesis of Example 2 YT-1

[0042] The synthetic route of YT-1 is the same as that of intermediate 1, using 4-pyridylboronic acid pinacol ester to replace 4-isopropylsulfonylphenylboronic acid pinacol ester and intermediate 2 to replace 5-bromo-7-azaindole. 1 HNMR (400 MHz, DMSO-d6) δ 12.42 (d, J = 2.8 Hz, 1H), 8.71 (s, 2H), 8.56 (d, J = 5.5 Hz, 2H), 8.29 (d, J = 2.7 Hz, 1H), 8.12 (d, J = 8.3 Hz, 2H), 7.94 (d, J = 8.4 Hz, 2H), 7.89–7.84 (m, 2H), 3.46 (p, J = 6.7 Hz, 1H), 1.19 (d, J = 6.7 Hz, 6H). 13 CNMR (101 MHz, DMSO-d6) δ 150.46, 149.80, 144.54, 143.16, 142.58, 135.64, 129.70, 128.46, 128.10, 128.03, 127.07, 121.05, 117.54, 112.58, 54.69, 15.73.

[0043] Synthesis of Example 3 YT-2

[0044] The synthesis route of YT-2 is the same as that of Intermediate 1, using 2-amino-5-pyrimidineboronic acid pinacol ester to replace 4-isopropylsulfonylphenylboronic acid pinacol ester and Intermediate 2 to replace 5-bromo-7-azaindole. 1 H NMR(400MHz,DMSO-d6)δ12.06(d,J=2.6Hz,1H),8.67(d,J=4.9Hz,3H),8.50(d,J=2.2Hz,1H),8.11(d,J=8.1Hz,2H),7.98–7.82(m,3H),6.62(s,2H),3.45(p,J=6.9Hz,1H),1.19(d,J=6.7Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ151.24,150.79,148.17,143.90,140.84,139.56,136.87,134.04,129.93,128.09,120.82,120.56,110.93,54.61,15.67.

[0045] Synthesis of Example 4 YT-3

[0046] The synthesis route of YT-3 is the same as that of Intermediate 1, using 2-methoxy-4-pyridineboronic acid pinacol ester to replace 4-isopropylsulfonylphenylboronic acid pinacol ester and Intermediate 2 to replace 5-bromo-7-azaindole. 1 H NMR(400MHz,DMSO-d6)δ12.38(s,1H),8.69(d,J=2.0Hz,1H),8.63(d,J=2.1Hz,1H),8.27(s,1H),8.17(d,J=5.4Hz,1H),8.12(d,J=8.2Hz,2H),7.94(d,J=8.1Hz,2H),7.48(dd,J=5.5,1.4Hz,1H),7.24(s,1H),3.89(s,3H),3.47(p,J=6.8Hz,1H),1.20(d,J=6.8Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ164.90,149.72,147.65,145.55,144.57,143.12,135.61,129.72,128.47,128.16,128.00,127.03,117.55,115.39,112.63,106.71,54.69,53.48,15.74.

[0047] Synthesis of Example 5 YT-4

[0048] The synthetic route of YT-4 is the same as that of Intermediate 1. Replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 3-pyridineboronic acid pinacol ester, and replace 5-bromo-7-azaindole with Intermediate 2. 1 HNMR(400MHz,DMSO-d6)δ12.27(d,J=2.7Hz,1H),9.06(s,1H),8.70(d,J=2.1Hz,1H),8.61(d,J=2.2Hz,1H),8.48(d,J=4.7Hz,1H),8.24(dt,J=8.0,2.0Hz,1H),8.15–8.08(m,3H),7.92(d,J=8.3Hz,2H),7.47(dd,J=8.0,4.7Hz,1H),3.45(p,J=6.8Hz,1H),1.19(d,J=6.9Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ149.61,147.83,147.28,144.64,142.99,135.52,134.02,131.17,129.70,128.37,127.68,126.78,126.36,124.47,117.70,112.12,54.69,15.73.

[0049] Example 6 Synthesis of YT-5

[0050] The synthetic route of YT-5 is the same as that of Intermediate 1. Replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 2-fluoro-3-pyridineboronic acid pinacol ester, and replace 5-bromo-7-azaindole with Intermediate 2. 1 HNMR(400MHz,DMSO-d6)δ12.38(s,1H),8.71(d,J=2.1Hz,1H),8.49(s,1H),8.41(ddd,J=9.9,7.4,1.9Hz,1H),8.16(d,J=4.8Hz,1H),8.12–8.04(m,2H),7.99(t,J=2.3Hz,1H),7.93(d,J=8.1Hz,2H),7.46(ddd,J=7.2,4.9,2.0Hz,1H),3.50–3.43(m,1H),1.19(d,J=6.8Hz,6H). 1313C NMR (101 MHz, DMSO-d6) δ 149.21, 144.86, 144.72, 144.57, 143.12, 140.60, 140.56, 135.60, 130.99, 129.75, 128.51, 128.44, 128.32, 127.69, 127.03, 127.00, 126.46, 123.03, 123.00, 118.07, 116.82, 107.29, 54.68, 15.73.

[0051] Synthesis of Example 7 YT-6

[0052] The synthesis route of YT-6 is the same as that of Intermediate 1. Replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 2-ethylamino-5-pyrimidineboronic acid pinacol ester, and replace 5-bromo-7-azaindole with Intermediate 2. 1 1H NMR (400 MHz, DMSO-d6) δ 12.06 (s, 1H), 8.73 (s, 2H), 8.68 (t, J = 1.7 Hz, 1H), 8.53 (d, J = 2.2 Hz, 1H), 8.17–8.09 (m, 2H), 7.95–7.89 (m, 2H), 7.88 (t, J = 1.9 Hz, 1H), 7.15 (t, J = 5.7 Hz, 1H), 3.46 (p, J = 6.8 Hz, 1H), 3.36 (d, J = 6.9 Hz, 2H), 1.22–1.18 (m, 6H), 1.16 (td, J = 7.1, 1.3 Hz, 3H). 13 13C NMR (101 MHz, DMSO-d6) δ 160.15, 148.91, 146.15, 144.09, 140.90, 136.49, 136.43, 130.63, 128.71, 125.68, 125.46, 122.28, 121.94, 115.39, 54.38, 36.16, 16.23, 15.76.

[0053] Synthesis of Example 8 YT-7

[0054] The synthesis route of YT-7 is the same as that of Intermediate 1. Replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 6-quinolineboronic acid pinacol ester, and replace 5-bromo-7-azaindole with Intermediate 2. 11H NMR (400 MHz, DMSO-d6) δ 12.28 (s, 1H), 8.84 (dd, J = 4.2, 1.7 Hz, 1H), 8.78 (d, J = 2.1 Hz, 1H), 8.71 (d, J = 2.0 Hz, 1H), 8.51–8.45 (m, 1H), 8.43 (d, J = 2.0 Hz, 1H), 8.24 (dd, J = 8.8, 2.1 Hz, 1H), 8.18 (d, J = 2.5 Hz, 1H), 8.13 (d, J = 8.4 Hz, 2H), 8.07 (d, J = 8.8 Hz, 1H), 7.95 (d, J = 8.4 Hz, 2H), 7.53 (dd, J = 8.3, 4.2 Hz, 1H), 3.48 (q, J = 6.8 Hz, 1H), 1.20 (d, J = 6.7 Hz, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 150.15, 149.76, 146.98, 144.79, 143.00, 136.41, 135.56, 133.36, 129.86, 129.73, 129.55, 129.10, 128.45, 127.76, 127.04, 126.72, 124.19, 122.10, 117.83, 114.73, 54.69, 15.74.

[0055] Synthesis of Example 9 YT-8

[0056] The synthetic route of YT-8 is the same as that of Intermediate 1, using indole-6-boronic acid pinacol ester to replace 4-isopropylsulfonylphenylboronic acid pinacol ester and Intermediate 2 to replace 5-bromo-7-azaindole. 1 1H NMR (400 MHz, DMSO-d6) δ 12.01 (s, 1H), 11.02 (s, 1H), 8.66 (d, J = 2.1 Hz, 1H), 8.55 (d, J = 2.2 Hz, 1H), 8.07 (d, J = 8.4 Hz, 2H), 7.93 (d, J = 8.4 Hz, 2H), 7.86 (s, 1H), 7.75 (s, 1H), 7.61 (d, J = 8.2 Hz, 1H), 7.41 (dd, J = 8.2, 1.5 Hz, 1H), 7.33 (t, J = 2.7 Hz, 1H), 6.43 (t, J = 2.4 Hz, 1H), 3.46 (s, 1H), 1.23–1.16 (m, 6H). 1313C NMR (101 MHz, DMSO-d6) δ 149.62, 144.98, 142.51, 137.08, 135.44, 129.79, 128.17, 128.02, 127.16, 126.67, 126.56, 125.86, 124.60, 120.93, 119.21, 118.22, 117.01, 109.80, 101.49, 54.69, 15.74.

[0057] Synthesis of Example 10 YT-9

[0058] The synthesis route of YT-9 is the same as that of Intermediate 1. Replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 7-azaindole-5-boronic acid pinacol ester, and replace 5-bromo-7-azaindole with Intermediate 2. 1 1H NMR (400 MHz, DMSO-d6) δ 12.08 (d, J = 2.7 Hz, 1H), 11.64 (s, 1H), 8.68 (d, J = 2.1 Hz, 1H), 8.63 (d, J = 2.1 Hz, 1H), 8.57 (d, J = 2.2 Hz, 1H), 8.33 (d, J = 2.1 Hz, 1H), 8.10 (d, J = 8.2 Hz, 2H), 7.92 (dd, J = 9.5, 2.6 Hz, 3H), 7.48 (t, J = 2.9 Hz, 1H), 6.49 (dd, J = 3.4, 1.8 Hz, 1H), 3.46 (q, J = 6.7 Hz, 1H), 1.19 (d, J = 6.7 Hz, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 146.77, 146.15, 144.09, 143.41, 140.90, 136.98, 136.49, 132.09, 130.76, 130.63, 128.71, 125.68, 125.46, 122.28, 119.59, 119.24, 113.26, 108.03, 54.38, 15.73.

[0059] Synthesis of Example 11 YT-10

[0060] The synthesis route of YT-9 is the same as that of Intermediate 1. Replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 7-azaindole-4-boronic acid pinacol ester, and replace 5-bromo-7-azaindole with Intermediate 2. 11H NMR (400 MHz, DMSO-d6) δ 12.38 (s, 1H), 11.69 (s, 1H), 8.70 (d, J = 2.2 Hz, 1H), 8.53 (d, J = 2.2 Hz, 1H), 8.25 (d, J = 5.0 Hz, 1H), 8.12 (d, J = 2.5 Hz, 1H), 8.05 (d, J = 8.1 Hz, 2H), 7.90 (d, J = 8.0 Hz, 2H), 7.52–7.42 (m, 2H), 6.68 (d, J = 2.8 Hz, 1H), 3.43 (q, J = 6.8 Hz, 2H), 1.16 (d, J = 6.8 Hz, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 149.63, 149.54, 144.68, 143.49, 143.12, 135.58, 134.73, 129.79, 128.29, 127.85, 127.65, 127.12, 126.21, 118.24, 117.36, 114.39, 113.09, 100.24, 54.68, 15.73.

[0061] Synthesis of Intermediate 3-5 in Example 12

[0062]

[0063] 1. Reagents and Conditions

[0064] (a) DMAP, TsCl, TEA, CH3CN, rt; (b) DMF, Et3N, CuI, PdCl2(PPh3)2, TMSA, overnight; (c) Na2CO3, MeOH, rt, 10 min; (d) DMF, Et3N, 60 °C, 1 h, then NaOH, MeOH, 1 h, rt.

[0065] 2. Synthesis of Intermediate 3

[0066] To a 50 mL round-bottom flask was added Intermediate 2 (460 mg, 1.1 mmol) and 4-dimethylaminopyridine (DMAP) (53.7 mg, 0.44 mmol). The mixture was dissolved in acetonitrile (25 mL) and stirred for 5 minutes. Then triethylamine (213 μL, 1.65 mmol) and p-toluenesulfonyl chloride (228 mg, 1.2 mmol) were added. The reaction was carried out at room temperature for 3 hours, and the reaction progress was monitored by thin-layer chromatography (TLC). After the reaction was completed, the reaction mixture was transferred to a 150 mL round-bottom flask and concentrated under reduced pressure. Subsequently, purification was carried out by flash column chromatography to obtain a dark yellow solid (388.51 mg, yield 61%). 11H NMR (400 MHz, DMSO-d6) δ 8.78 - 8.75 (m, 1H), 8.22 (d, J = 1.0 Hz, 1H), 8.03 (d, J = 2.2 Hz, 3H), 8.01 (s, 1H), 7.91 (d, J = 8.4 Hz, 2H), 7.42 (d, J = 8.1 Hz, 2H), 3.45 (p, J = 6.8 Hz, 1H), 2.32 (s, 3H), 1.15 (dd, J = 6.8, 1.1 Hz, 6H).

[0067] 3. Synthesis of Intermediate 4

[0068] To a 150 mL three-necked flask were added Intermediate 3 (1.91 g, 3.3 mmol), trimethylacetylene (511.7 μL, 3.6 mmol), triethylamine (1.07 mL, 8.25 mmol), copper(I) iodide (62.7 mg, 0.33 mmol), bis(triphenylphosphine)palladium(II) dichloride (231.6 mg, 0.33 mmol) and N,N-dimethylformamide (DMF, 25 mL). Under nitrogen protection, the mixture was reacted overnight at room temperature. After completion of the reaction, 125 mL of water was added to dilute the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and purified by flash column chromatography using a petroleum ether / ethyl acetate system to obtain 1.01 g of Intermediate 4 as a white powder, with a yield of 56%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.81 (d, J = 2.2 Hz, 1H), 8.34 (s, 1H), 8.24 (d, J = 2.2 Hz, 1H), 8.08–8.01 (m, 4H), 7.94 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 8.2 Hz, 2H), 3.47 (p, J = 6.8 Hz, 1H), 2.35 (s, 3H), 1.18 (d, J = 6.8 Hz, 6H), 0.27 (s, 9H).

[0069] Synthesis of Intermediate 5 in Example 13

[0070] To a 250 mL round-bottom flask were added Intermediate 4 (1.75 g, 3.18 mmol), methanol (75 mL) and sodium carbonate (848 mg, 8 mmol). After completion of the reaction, the solvent was removed under reduced pressure at room temperature. The collected solid was dissolved in a mixed solvent of dichloromethane and ethyl acetate, and then filtered. The filtrate was concentrated under reduced pressure to obtain 1.6 g of a white solid, with a yield of 98%. 1HNMR(400MHz, DMSO-d6) δ 8.83 (d, J = 2.2Hz, 1H), 8.36 (s, 1H), 8.33 (d, J = 2.2Hz, 1H), 8.09 - 8.03 (m, 4H), 7.95 - 7.90 (m, 2H), 7.45 (d, J = 8.3Hz, 2H), 4.51 (s, 1H), 3.47 (p, J = 6.7Hz, 1H), 2.35 (s, 3H), 1.18 (d, J = 6.8Hz, 6H).

[0071] Synthesis of Example 14 YT-11

[0072] Add intermediate 5 (239 mg, 0.5 mmol), α-chlorobenzaldoxime (116.3 mg, 0.75 mmol) and N,N-dimethylformamide (10 mL) to a 25 mL round-bottom flask. Heat the reaction mixture to 60 °C, and then add triethylamine (161 μL, 1.25 mmol) dropwise. Monitor the reaction progress by thin-layer chromatography (TLC), and the reaction lasts for two hours. After the reaction is completed, cool the mixture to room temperature and then transfer it to a flask containing 30 mL of water. The resulting solution is extracted three times with an equal volume of ethyl acetate. Combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and then concentrate under reduced pressure to obtain a white solid. Dissolve the intermediate product in methanol and add saturated sodium hydroxide solution dropwise until the alkaline condition is reached. After stirring at room temperature for 1 hour, remove the solvent under reduced pressure. Purify the crude product by flash column chromatography to obtain a pure white solid product with a yield of 27%. 1 H NMR(400MHz, DMSO-d6) δ 12.63 (s, 1H), 8.77 (s, 2H), 8.32 (s, 1H), 8.16 (d, J = 1.9Hz, 1H), 8.14 (d, J = 2.0Hz, 1H), 8.01 (d, J = 1.9Hz, 1H), 7.99 (d, J = 2.0Hz, 2H), 7.98 (d, J = 1.8Hz, 1H), 7.58 (d, J = 3.3Hz, 2H), 7.57–7.51 (m, 2H), 3.50 (p, J = 6.8Hz, 1H), 1.22 (d, J = 6.8Hz, 6H). 13 C NMR(101MHz, DMSO-d6) δ 166.31, 162.80, 149.20, 144.29, 143.82, 135.82, 130.64, 129.76, 129.54, 129.37, 128.58, 128.18, 127.18, 126.99, 116.61, 103.35, 96.90, 54.70, 15.74.

[0073] Synthesis of Example 15 YT-12

[0074] The synthesis route of YT-12 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldehyde oxime with 4-fluoro-α-chlorobenzaldehyde oxime. 1 HNMR(400MHz,DMSO-d6)δ12.65(s,1H),8.77(s,2H),8.32(s,1H),8.14(d,J=8.2Hz,2H),8.09–8.03(m,2H),7.99(d,J=8.2Hz,2H),7.58(s,1H),7.42(t,J=8.8Hz,2H),3.50(p,J=6.8Hz,1H),1.22(d,J=6.8Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ166.38,164.89,162.43,161.92,149.16,144.28,143.84,135.84,129.76,129.52,129.43,128.61,128.58,128.15,126.97,125.95,125.92,116.70,116.58,116.48,103.28,96.90,54.70,15.73.

[0075] Synthesis of Example 16 YT-13

[0076] The synthesis route of YT-13 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldehyde oxime with 4-methyl-α-chlorobenzaldehyde oxime. 1 HNMR(400MHz,DMSO-d6)δ8.77(s,2H),8.30(s,1H),8.20–8.11(m,3H),8.01–7.96(m,2H),7.89(d,J=8.1Hz,2H),7.54(s,1H),7.37(d,J=8.0Hz,2H),5.76(s,1H),4.08–3.96(m,1H),2.39(s,3H),1.22(d,J=6.8Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ166.12,162.69,149.20,144.30,143.79,140.31,135.81,130.07,129.75,128.58,128.12,127.08,126.99,126.57,116.61,103.40,96.79,54.69,21.47,15.73.

[0077] Synthesis of Example 17 YT-14

[0078] The synthetic route of YT-14 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldoxime with 4-chloro-α-chlorobenzaldoxime. 1 HNMR(400MHz,DMSO-d6)δ8.76(s,2H),8.28(s,1H),8.22–8.10(m,3H),8.00–7.95(m,2H),7.90(d,J=8.1Hz,2H),7.54(s,1H),7.37(d,J=8.0Hz,2H),5.76(s,1H),4.08–3.96(m,1H),2.39(s,3H),1.22(d,J=6.8Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ157.55,154.98,146.22,144.19,140.93,138.28,136.13,132.01,130.63,129.88,128.77,128.70,128.26,125.58,124.43,121.50,111.40,106.48,54.38,15.53.

[0079] Example 18 Synthesis of YT-15

[0080] The synthetic route of YT-15 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldoxime with 3-amino-α-chlorobenzaldoxime. 1 HNMR(400MHz,DMSO-d6)δ9.14(d,J=3.1Hz,1H),8.48(d,J=2.9Hz,1H),8.12(s,1H),7.99(s,4H),7.56(s,1H),7.44(t,J=14.8Hz,1H),7.30(dt,J=15.2,3.1Hz,1H),6.89(t,J=2.9Hz,1H),6.74(dt,J=14.7,3.1Hz,1H),4.26(s,2H),4.10–3.98(m,1H),1.26(d,J=13.1Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ160.95,160.41,145.69,144.18,141.92,139.87,139.38,138.30,134.06,133.22,129.43,128.64,127.03,126.37,122.20,116.10,115.15,113.58,106.68,101.44,55.47,16.27,-19.86.

[0081] Synthesis of Example 19 YT-16

[0082] The synthesis route of YT-16 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldehyde oxime with 4-amino-α-chlorobenzaldehyde oxime. 1 HNMR(400MHz,DMSO-d6)δ9.34(d,J=3.1Hz,1H),8.56(d,J=2.9Hz,1H),8.32(s,1H),8.01(s,4H),7.78(s,1H),7.68–7.57(m,2H),6.76–6.40(m,2H),5.24(s,2H),4.05–3.93(m,1H),1.32(d,J=13.1Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ160.99,160.95,149.01,144.18,141.92,139.87,139.38,138.30,133.22,128.64,127.18,127.03,126.37,116.31,114.35,113.58,106.68,101.44,55.47,16.27,-19.79.

[0083] Synthesis of Example 20 YT-17

[0084] The synthesis route of YT-17 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldehyde oxime with 4-aminomethyl-α-chlorobenzaldehyde oxime. 1 HNMR(400MHz,DMSO-d6)δ9.88(s,1H),8.89(d,J=2.0Hz,1H),8.81(d,J=2.1Hz,1H),8.69(s,1H),8.13(d,J=4.0Hz,2H),8.00(d,J=2.5Hz,2H),7.87(s,1H),7.65(d,J=8.3Hz,2H),7.48(d,J=8.2Hz,2H),4.84(d,J=11.3Hz,2H),3.51(dt,J=13.6,6.8Hz,1H),1.21(d,J=1.4Hz,6H).

[0085] Synthesis of Example 21 YT-18

[0086] The synthesis route of YT-18 is the same as that of YT-11, and it is only necessary to replace α-chlorobenzaldehyde oxime with 4-methylaminomethyl-α-chlorobenzaldehyde oxime. 11H NMR (400 MHz, DMSO-d6) δ 11.83 (s, 1H), 9.24 (d, J = 3.1 Hz, 1H), 8.54 (d, J = 2.9 Hz, 1H), 8.12 (s, 1H), 8.04–7.93 (m, 4H), 7.72–7.64 (m, 2H), 7.51–7.43 (m, 2H), 7.39 (s, 1H), 3.76 (s, 2H), 3.47 (dt, J = 26.1, 13.2 Hz, 1H), 3.26 (s, 3H), 1.46 (d, J = 13.2 Hz, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 160.99, 160.95, 144.18, 141.92, 139.87, 139.38, 138.30, 137.40, 133.22, 130.00, 128.64, 127.77, 127.03, 126.80, 126.37, 113.58, 106.68, 101.44, 55.56, 55.47, 36.63, 16.27.

[0087] Synthesis of Example 22 YT-19

[0088] The synthetic route of YT-19 is the same as that of YT-11, and 4-cyclopropylaminomethyl-α-chlorobenzaldoxime can be used instead of α-chlorobenzaldoxime. 1 1H NMR (400 MHz, DMSO-d6) δ 8.77 (s, 2H), 8.31 (s, 1H), 8.15 (d, J = 8.5 Hz, 2H), 7.99 (d, J = 8.4 Hz, 3H), 7.93 (d, J = 8.2 Hz, 2H), 7.56 (s, 1H), 7.52–7.46 (m, 2H), 3.80 (s, 2H), 3.52–3.46 (m, 2H), 2.06 (tt, J = 6.7, 3.6 Hz, 1H), 1.22 (d, J = 6.8 Hz, 6H), 0.36 (tt, J = 6.4, 3.6 Hz, 2H), 0.32–0.23 (m, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 170.68, 166.17, 162.71, 149.18, 144.29, 143.92, 143.79, 135.81, 129.84, 129.75, 129.01, 128.56, 128.40, 128.32, 128.11, 127.52, 127.42, 126.98, 126.90, 116.61, 103.40, 96.85, 54.70, 53.07, 30.41, 15.73, 6.65.

[0089] Synthesis of Intermediates 6 and 7 in Example 23

[0090]

[0091] 1. Reagents and Conditions

[0092] (a) PdCl2(dppf), K2CO3, Dioxane / H2O / EtOH, 85 °C, 4 h; (b) THF, NIS, rt, 3 h; (c) DMAP, TsCl, NEt3, CH3CN, rt, 3 h; (d) PdCl2(dppf), K2CO3, Dioxane / H2O / EtOH, 85 °C, 4 h.

[0093] 2. Synthesis of Intermediate 6

[0094] The synthesis route of Intermediate 6 is the same as that of Intermediate 1, just replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 4-dimethylaminophenylboronic acid pinacol ester. 1 H NMR(400MHz, DMSO-d6) δ11.62(s,1H), 8.46(s,1H), 8.11(s,1H), 7.56(d,J = 8.0Hz,2H), 7.48(s,1H), 7.03(d,J = 8.1Hz,2H), 6.47(s,1H), 4.23(s,6H).

[0095] 3. Synthesis of Intermediate 7

[0096] The synthesis route of Intermediate 7 is the same as that of Intermediate 2, just replace Intermediate 1 with Intermediate 6. 1 H NMR(400MHz, DMSO-d6) δ11.62(s,1H), 8.46(s,1H), 7.56(d,J = 8.0Hz,2H), 7.48(s,1H), 7.03(d,J = 8.1Hz,2H), 6.47(s,1H), 4.25(s,6H).

[0097] Synthesis of YT-20 in Example 24

[0098] The synthesis route of YT-20 is the same as that of Intermediate 1, just replace 4-isopropylsulfonylphenylboronic acid pinacol ester with 7-azaindole-4-boronic acid pinacol ester and replace 5-bromo-7-azaindole with Intermediate 7. 11H NMR (400 MHz, DMSO-d6) δ 12.16 (s, 1H), 11.69 (s, 1H), 8.56 (s, 1H), 8.41–8.19 (m, 2H), 8.05 (s, 1H), 7.66–7.38 (m, 4H), 6.84 (d, J = 8.2 Hz, 2H), 6.69 (d, J = 3.4 Hz, 1H), 2.94 (s, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 150.13, 149.65, 148.50, 143.47, 142.37, 135.22, 129.86, 128.01, 127.06, 126.04, 125.11, 118.24, 117.35, 114.21, 113.41, 112.59, 100.32, 40.60.

[0099] Synthesis of Example 25 YT-21

[0100] The synthetic route of YT-21 is the same as that of YT-20. Just replace 4-(dimethylamino)phenylboronic acid pinacol ester with 4-(4-methylpiperazin-1-yl)phenylboronic acid pinacol ester. 1 1H NMR (400 MHz, DMSO-d6) δ 12.20 (s, 1H), 11.68 (s, 1H), 8.60 (s, 1H), 8.34 (s, 2H), 8.06 (d, J = 2.4 Hz, 1H), 7.60 (d, J = 8.2 Hz, 2H), 7.52–7.43 (m, 2H), 7.04 (d, J = 8.3 Hz, 2H), 6.71 (s, 1H), 3.18 (t, J = 4.8 Hz, 4H), 2.47 (t, J = 4.8 Hz, 4H), 2.23 (s, 3H). 13 13C NMR (101 MHz, DMSO-d6) δ 150.57, 149.65, 148.64, 143.45, 142.45, 135.18, 129.62, 129.57, 128.56, 128.01, 127.18, 126.06, 125.41, 118.26, 117.40, 116.25, 114.27, 112.67, 100.32, 54.92, 48.34, 46.12.

[0101] Synthesis of Example 26 YT-22

[0102] The synthetic route of YT-22 is the same as that of YT-20. Just replace 4-(dimethylamino)phenylboronic acid pinacol ester with 4-(4-methylpiperazin-1-yl)phenylboronic acid pinacol ester and 7-azaindole-4-boronic acid pinacol ester with 7-azaindole-5-boronic acid pinacol ester. 1HNMR(400MHz,DMSO-d6)δ11.89(s,1H),11.64(s,1H),8.61(d,J=45.5Hz,2H),8.35(s,1H),8.31(s,1H),7.87(d,J=2.3Hz,1H),7.62(d,J=8.2Hz,2H),7.49(d,J=2.8Hz,1H),7.03(d,J=8.3Hz,2H),6.52(s,1H),3.17(t,J=4.6Hz,4H),2.47(d,J=9.7Hz,4H),2.23(s,3H). 13 C NMR(101MHz,DMSO-d6)δ150.49,148.65,147.76,142.06,129.86,129.27,128.04,126.94,126.21,124.77,124.22,123.44,120.40,118.22,116.23,113.52,100.43,54.94,48.40,46.14.

[0103] Synthesis of Example 27 YT-23

[0104] The synthetic route of YT-23 is the same as that of YT-20, and 7-azaindole-5-boronic acid pinacol ester can be used instead of 7-azaindole-4-boronic acid pinacol ester. 1 H NMR(400MHz,DMSO-d6)δ11.88(s,1H),11.63(s,1H),8.59(t,J=8.4Hz,1H),8.52(d,J=2.0Hz,1H),8.34(d,J=2.1Hz,1H),8.32(s,1H),8.29(t,J=6.4Hz,1H),7.85(d,J=9.7Hz,1H),7.62(d,J=8.7Hz,2H),7.03(t,J=7.4Hz,2H),6.54–6.46(m,1H),3.30(s,4H),3.17(d,J=4.7Hz,4H),2.31–2.25(m,2H),1.47(dt,J=14.6,7.3Hz,2H),0.88(t,J=7.4Hz,3H). 1313C NMR (101 MHz, DMSO-d6) δ 150.59, 148.64, 147.75, 142.10, 142.05, 129.83, 129.23, 128.05, 126.92, 126.17, 124.76, 124.20, 123.36, 120.32, 118.15, 116.19, 113.48, 100.40, 60.29, 53.16, 48.63, 19.96, 12.32.

[0105] Synthesis of Intermediates 8 and 9 in Example 28

[0106]

[0107] 1. Reagents and Conditions

[0108] (a) PdCl2(dppf), K2CO3, Dioxane / H2O / EtOH, 85 °C, 4 h, (b) THF, NIS, rt, 3 h; (c) PdCl2(dppf), K2CO3, Dioxane / H2O / EtOH, 85 °C, 4 h, (d) NaOH, DMF, 90 °C, 2 h.

[0109] 2. Synthesis of Intermediate 8

[0110] The synthesis of Intermediate 8 is the same as that of Intermediate 1, except that 2-fluoro-5-pyridylboronic acid pinacol ester is used instead of 4-isopropylsulfonylphenylboronic acid pinacol ester. 1 1H NMR (400 MHz, Chloroform-d) δ 9.67 (d, J = 6.4 Hz, 1H), 8.77 (dd, J = 3.6, 1.9 Hz, 1H), 8.67 (d, J = 1.8 Hz, 1H), 8.63 (t, J = 2.0 Hz, 1H), 8.13 (ddd, J = 8.1, 3.7, 2.0 Hz, 1H), 7.21–7.13 (m, 1H), 7.05 (dd, J = 6.4, 4.9 Hz, 1H), 6.80 (dd, J = 4.9, 2.2 Hz, 1H).

[0111] 3. Synthesis of Intermediate 9

[0112] The synthesis of Intermediate 9 is the same as that of Intermediate 2, except that Intermediate 8 is used instead of Intermediate 1. 11H NMR (400 MHz, Chloroform-d) δ 9.67 (d, J = 6.4 Hz, 1H), 8.77 (dd, J = 3.6, 1.9 Hz, 1H), 8.67 (d, J = 1.8 Hz, 1H), 8.63 (t, J = 2.0 Hz, 1H), 8.13 (ddd, J = 8.1, 3.7, 2.0 Hz, 1H), 7.21–7.13 (m, 1H), 6.89 (s, 1H).

[0113] Synthesis of Example 29 YT-24

[0114] The synthetic route of YT-24 is the same as that of YT-20, and intermediate 9 can be used instead of intermediate 7. 1 1H NMR (400 MHz, DMSO-d6) δ 12.36 (s, 1H), 11.70 (s, 1H), 8.66 (s, 2H), 8.51 (s, 1H), 8.46–8.37 (m, 1H), 8.28 (d, J = 4.9 Hz, 1H), 8.14 (d, J = 2.5 Hz, 1H), 7.50 (d, J = 5.4 Hz, 2H), 7.31 (dd, J = 8.4, 2.8 Hz, 1H), 6.79–6.66 (m, 1H). 13 13C NMR (101 MHz, DMSO-d6) δ 164.06, 161.72, 149.62, 149.28, 146.16, 146.01, 143.47, 142.76, 141.28, 141.20, 134.80, 133.62, 133.57, 127.74, 126.89, 126.16, 125.31, 118.20, 117.33, 114.38, 112.92, 110.26, 109.89, 100.27.

[0115] Synthesis of Example 30 YT-25

[0116] Add compound YT24 (150 mg, 0.31 mmol), morpholine (54 mg, 0.61 mmol), sodium hydroxide (32 mg, 0.81 mmol) and N,N-dimethylformamide (DMF, 20 mL) to a 50 mL single-necked flask, and react at 90 °C for 2 hours. After the reaction is completed, add 40 mL of water and dichloromethane (40 mL) to extract the mixture. The organic phase is concentrated under reduced pressure and then purified by flash column chromatography to obtain the target product with a yield of 75%. 11H NMR (400 MHz, DMSO-d6) δ 12.24 (s, 1H), 11.69 (s, 1H), 8.56 (dd, J = 15.7, 2.3 Hz, 2H), 8.38 (d, J = 2.1 Hz, 1H), 8.27 (d, J = 4.9 Hz, 1H), 8.09 (s, 1H), 7.98 (dd, J = 8.8, 2.6 Hz, 1H), 7.48 (dd, J = 17.3, 4.3 Hz, 2H), 6.96 (d, J = 8.9 Hz, 1H), 6.71 (d, J = 3.5 Hz, 1H), 3.73 (t, J = 4.8 Hz, 4H), 3.50 (t, J = 4.8 Hz, 4H). 13 13C NMR (101 MHz, DMSO-d6) δ 158.79, 149.63, 148.78, 146.18, 143.49, 142.24, 136.77, 135.04, 127.35, 126.98, 126.08, 125.56, 124.81, 118.26, 117.31, 114.29, 112.67, 107.61, 100.30, 66.42, 45.70.

[0117] Synthesis of Example 31 YT-26

[0118] The synthetic route of YT-26 is the same as that of YT-25, and N-propylpiperazine can be used instead of morpholine. 1 1H NMR (400 MHz, DMSO-d6) δ 12.26 (s, 1H), 11.78 (s, 1H), 8.50 (dd, J = 10.1, 2.1 Hz, 2H), 8.31 (s, 1H), 8.20 (d, J = 5.2 Hz, 1H), 8.07 (d, J = 2.5 Hz, 1H), 7.96 (dd, J = 8.8, 2.5 Hz, 1H), 7.45 (d, J = 4.7 Hz, 2H), 6.99 (d, J = 8.8 Hz, 1H), 6.68 (d, J = 3.5 Hz, 1H), 4.36 (d, J = 13.6 Hz, 2H), 3.49 (d, J = 11.7 Hz, 2H), 3.04 (dt, J = 29.0, 10.5 Hz, 6H), 1.66–1.53 (m, 2H), 0.83 (t, J = 7.4 Hz, 3H). 13CNMR(101MHz, DMSO-d6) δ 157.60, 149.59, 148.83, 146.18, 143.47, 142.29, 137.12, 135.03, 127.43, 126.75, 126.12, 125.68, 125.50, 118.26, 117.33, 114.31, 112.70, 108.28, 100.30, 57.92, 51.26, 42.91, 17.61, 11.43.

[0119] Synthesis of Example 32 YT-27

[0120] The synthetic route of YT-27 is the same as that of YT-25, and 3-(4-morpholino)-1-propanol can be used to replace morpholine. 1 H NMR(400MHz, DMSO-d6) δ 12.29(s, 1H), 11.69(s, 1H), 8.61(t, J = 1.7Hz, 1H), 8.54(t, J = 1.7Hz, 1H), 8.45–8.39(m, 1H), 8.28(dd, J = 4.9, 1.3Hz, 1H), 8.11(ddt, J = 5.2, 3.9, 1.8Hz, 2H), 7.53–7.49(m, 1H), 7.47(dd, J = 5.0, 1.3Hz, 1H), 6.92(d, J = 8.5Hz, 1H), 6.71(dt, J = 3.4, 1.5Hz, 1H), 4.38–4.31(m, 2H), 3.61–3.54(m, 4H), 2.47–2.32(m, 6H), 1.91(p, J = 6.8Hz, 2H). 13 C NMR(101MHz, DMSO-d6) δ 163.09, 149.63, 149.01, 145.35, 143.47, 142.49, 138.47, 134.97, 128.51, 127.49, 126.45, 126.19, 126.10, 118.24, 117.34, 114.32, 112.79, 111.20, 100.29, 66.63, 64.47, 55.38, 53.76, 26.10.

[0121] Synthesis of Example 33 YT-28

[0122] The synthetic route of YT-28 is the same as that of YT-25, and 3-dimethylamino-1-propanol can be used to replace morpholine. 11H NMR (400 MHz, DMSO-d6) δ 12.29 (s, 1H), 11.69 (s, 1H), 8.61 (t, J = 1.7 Hz, 1H), 8.54 (d, J = 2.5 Hz, 1H), 8.42 (d, J = 2.1 Hz, 1H), 8.28 (d, J = 4.9 Hz, 1H), 8.11 (td, J = 4.9, 3.6, 1.9 Hz, 2H), 7.50 (d, J = 2.9 Hz, 1H), 7.47 (dd, J = 5.0, 1.3 Hz, 1H), 6.92 (d, J = 8.6 Hz, 1H), 6.71 (dd, J = 3.5, 1.8 Hz, 1H), 4.34 (t, J = 6.5 Hz, 2H), 2.44 (t, J = 7.2 Hz, 2H), 2.21 (s, 6H), 1.90 (p, J = 6.8 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 163.08, 149.61, 148.98, 145.36, 143.47, 142.49, 138.55, 134.93, 128.55, 127.50, 126.42, 126.19, 126.12, 118.22, 117.31, 114.32, 112.76, 111.22, 100.27, 64.35, 56.07, 45.34, 26.89.

[0123] Synthesis of Example 34 YT-29

[0124] The synthetic route of YT-29 is the same as that of YT-25, just replace morpholine with 1-piperidinepropanol. 1 1H NMR (400 MHz, DMSO-d6) δ 12.32 (s, 1H), 11.72 (s, 1H), 8.62 (d, J = 2.1 Hz, 1H), 8.55 (d, J = 2.5 Hz, 1H), 8.44 (d, J = 2.2 Hz, 1H), 8.29 (d, J = 5.0 Hz, 1H), 8.11 (d, J = 8.7 Hz, 2H), 7.62–7.41 (m, 2H), 6.91 (d, J = 8.5 Hz, 1H), 6.73 (d, J = 4.1 Hz, 1H), 4.32 (t, J = 6.6 Hz, 2H), 2.48–2.11 (m, 6H), 1.89 (p, J = 6.8 Hz, 2H), 1.49 (p, J = 5.6 Hz, 4H), 1.37 (q, J = 5.8 Hz, 2H). 1313C NMR(101MHz, DMSO-d6) δ 163.13, 149.62, 148.99, 145.37, 143.47, 142.49, 138.51, 134.93, 128.50, 127.49, 126.44, 126.19, 126.10, 118.21, 117.31, 114.31, 112.76, 111.22, 100.27, 64.66, 55.70, 54.53, 26.55, 26.03, 24.58.

[0125] Example 35

[0126] Antitumor proliferation experiment of combined use of chemotherapeutic drugs in vitro

[0127] HCT116 was seeded in 96-well plates at a density of 3×10 3 and 5×10 3 . In the combined group, after incubation with the ATM inhibitor for 1 h, different concentrations of irinotecan were added, and after incubation at 37 °C for 6 days, cell proliferation was measured by MTT.

[0128] Kinase activity assay

[0129] Eurofins provided services for the inhibition experiments of ATM, ATR and other kinase activities. Each kinase was incubated with the corresponding compound (at a specific concentration) in a specified reaction solution (with different pH and concentrations according to the specific requirements of different kinases). The reaction was initiated by adding a Mg(n) / ATP mixture. After incubation at room temperature for a certain period of time, the reaction was terminated by adding phosphoric acid at a concentration of 0.5%. 10 μL of the terminated reaction solution was spotted onto a P30 filter pad, washed 4 times with 0.425% phosphoric acid solution for 4 min each time, and finally washed once in methanol, followed by drying and scintillation counting. Detailed experimental procedures can be referred to the website https: / / www.eurofinsdiscoveryservices.com.

[0130] Table 1 In vitro cell activity and kinase activity of compounds

[0131]

[0132] Cell IC 50 (nM): # < 50; < 250; ## < 500; # < 1000.

[0133] Kinase IC 50 (nM): **** < 5; *** < 50; ** < 1000; * < 10000.

[0134] Table 2 Kinase selectivity of preferred compounds

[0135]

[0136] Kinase IC 50 (nM): ***** < 5; **** < 50; *** < 1000; ** < 5000; * < 10000.

[0137] Example 36

[0138] HCT116 cell colony formation assay

[0139] Collect HCT116 cells by centrifugation, then resuspend them in complete DMED medium and count. Place the cells in a 24-well plate at a density of 400 cells per well. Place the 24-well plate in an incubator containing 5% CO2 and culture at 37 °C for three days. Incubate the cells with the compound at a preset concentration for 1 hour, then irradiate the drug-treated cells with 2 Gy. After irradiation, place the cells in the incubator for one week. After one week, remove the cell supernatant and wash twice with PBS, then slowly add methanol and fix at room temperature for 20 minutes. After removing the methanol, add crystal violet to the dye and stain for 20 min, then wash it off with PBS. Subsequently, take photos of each well plate using a chemiluminescence imaging system, and use Image J to analyze the results of the colony formation experiment for quantitative analysis, and finally use it to calculate the corresponding inhibition rate and IC 50 value.

[0140] The experimental results are as Figure 1 shown:

[0141] 1. Figure 1 A - B shows that when compound YT-11 is combined with irinotecan, it can significantly enhance the inhibitory effect of irinotecan on these two cell lines in a concentration-dependent manner. In HCT116 cells, the combination of 1 μM YT-11 and irinotecan reduced the half-maximal inhibitory concentration (IC 50 ) value of irinotecan by approximately 7-fold, which is comparable to the effect of 2 μM positive control drug AZ32. Similarly, in SW620 cells, when two different concentrations of compound YT-11 were combined with irinotecan respectively, the IC 50 values of irinotecan were reduced by approximately 3-fold and 40-fold, respectively.

[0142] 2. The colony formation assay shows that as the concentration of irinotecan increases, the colony formation rate decreases ( Figure 1 C - F). The combination of AZ32 and compound YT-11 with irinotecan further inhibits the proliferation of tumor cells.

[0143] In summary, compound YT-11 significantly enhances the responsiveness of different cancer cell lines to chemotherapy.

[0144] Example 37

[0145] Western blot analysis experiment

[0146] HCT116 and SW620 cells were seeded in 6-well plates at a density of 3×10 4 cells per well. MCF-7 cells were harvested 4 hours after compound treatment and lysed in RIPA lysis buffer containing 10 mM phenylmethylsulfonyl fluoride (PMSF). Equal amounts of total protein lysates were separated on an SDS-PAGE electrophoresis apparatus and transferred to a PVDF membrane, which was then blocked with 5% non-fat milk for 2 hours, and the corresponding phosphorylated primary antibodies were added respectively. Then the blots were washed three times with PBST buffer and incubated with a goat anti-rabbit IgG HRP-labeled secondary antibody (1:5000 dilution; Earthox) for 1 hour. After washing three times with TBS-T, the blots were soaked with SuperECL Plus hypersensitive luminescent solution and observed by a chemiluminescence imager.

[0147] γH2AX immunofluorescence analysis experiment

[0148] HCT116 cells were cultured in 6-well plates containing round glass coverslips at a density of 5×10 3 for 48 hours, then different concentrations of compounds were added, and then 25 μM irinotecan was added to the growth medium. After incubation for 6 hours, the cells were washed three times with cold PBS, then fixed in cold MeOH for 15 minutes, permeabilized with Triton X-100 (0.2%) for 5 minutes, and blocked in BSA (2.5%) for 1 hour. Subsequently, the cells were immunostained with a monoclonal rabbit anti-γH2AX antibody respectively before incubation with a Fluor 594-conjugated goat anti-rabbit antibody. Finally, the round glass coverslip was fixed with a fixing solution containing Hoechst and observed and recorded using a fluorescence microscope for imaging.

[0149] The experimental results are as shown in the appendix Figure 2 as follows:

[0150] 1. As shown in 2A-B, irinotecan (10 μM) induced the high expression of the DSB damage marker γH2AX, while after adding the positive controls AZ32 or YT-11, the expression of γH2AX was downregulated. Without affecting the total ATM expression, irinotecan led to the upregulation of p-ATM (S1981), suggesting the activation of the ATM signaling pathway; after adding AZ32 or YT-11, the degree of phosphorylated ATM expression decreased significantly, and the inhibitory effect of YT-11 was more obvious at the same concentration. Generally speaking, compound YT-11 has an obvious inhibitory effect on the ATM-p53-p21 signaling pathway.

[0151] 2. The widespread expression of γH2AX (red fluorescence), and as the concentration of YT-11 increases, the intensity of red fluorescence gradually weakens, indicating that the combination of ATM inhibitor and irinotecan inhibits the activation of the ATM signaling pathway in tumor cells within a short time ( Figure 2 C).

[0152] Example 38

[0153] SW620 BALB / c nude mouse xenograft model

[0154] In this example, 6-week-old BALB / c nude mice from Beijing Huafukang Bioscience Co., Ltd. were used. SW620 cells were collected, rinsed twice with serum-free medium, resuspended, and subcutaneously injected into the right axillary region of the mice (about 1×10 7 cells, total volume 0.1 mL). When the tumor volume of each nude mouse reached 100 - 150 mm 3 , the animals were randomly divided into different treatment groups and control groups (5 animals / group). The liposomal irinotecan in the administration group was given once a day (2 mg / kg); the ATM inhibitor was given once a day (30 mg / kg). The activities, physical conditions, body weights, and tumor growth of all animals were monitored. The tumor size was calculated by measuring two perpendicular diameters of the tumor mass with a vernier caliper every three days. The tumor volume (unit: mm 3 ) was calculated by the formula (a×b 2 ) / 2, where "a" is the long diameter and "b" is the short diameter (unit: mm). After three weeks, the mice were sacrificed and the tumor tissues were collected. The animal studies in this invention were conducted in accordance with the institutional guidelines for the care and use of laboratory animals, and all mouse operation procedures were approved by the Animal Care and Use Committee of Sichuan University.

[0155] The experimental results are as shown in the appendix Figure 3 as follows:

[0156] The tumor growth inhibition rate (TGI) of liposomal irinotecan (2 mg / kg) reached 58.4%. The combination treatment of YT-11 (30 mg / kg) and liposomal irinotecan showed a dose-dependent enhanced inhibitory effect, and the tumor growth inhibition rate reached 95.4%. The results indicate that in cancer treatment, the combination of YT-11 and irinotecan has a synergistic anti-tumor effect.

[0157] The description and drawings of this invention are considered illustrative rather than restrictive. Based on this invention, those skilled in the art can make some substitutions and deformations of some technical features without creative labor according to the disclosed technical content, and all are within the protection scope of this invention.

Claims

1. A class of compounds containing a disubstituted 7-azaindole structure, characterized in that, It is a compound having a structure shown in formula (Ι), or a salt of the compound, or a solvate of the compound; wherein, R1 is selected from one of hydrogen, halogen, amino, cyano, acyl, sulfonyl, ureido, or substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkyl, five-membered or six-membered nitrogenous saturated alkyl; Ring A is selected from one of a substituted or unsubstituted benzene ring, a substituted or unsubstituted nitrogen-containing heterocycle, and a substituted or unsubstituted aromatic fused ring.

2. A compound of a class containing a 7-azaindole structure with a disubstitution according to claim 1, characterized in that, The A ring is selected from one of the following.

3. A compound of a class containing a disubstituted 7-azaindole structure according to claim 2, characterized in that, The pyridine in the ring A is disubstituted at the 2,5 positions, and the benzene ring is para-substituted.

4. A compound containing a disubstituted 7-azaindole structure according to claim 1, characterized in that, The R1 is a substituted C1-C5 alkoxy or a six-membered nitrogenous saturated alkyl.

5. A compound of the class containing a 7-azaindole structure with a disubstituted group according to claim 1, characterized in that, The compound is one of the following structures:

6. Use of a class of compounds containing a disubstituted 7-azaindole structure as described in any one of claims 1-5 in the preparation of a drug for preventing or treating ATM-mediated diseases.

7. The application according to claim 6, characterized in that, The disease is colorectal cancer.

8. The application according to claim 6, wherein The compound is used in combination with liposomal irinotecan.

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