Proliferating cell nuclear antigen inhibitor, and preparation method therefor, intermediate thereof, pharmaceutical composition thereof and use thereof

Abstract

A compound represented by formula (I) as a PCNA inhibitor, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, a preparation method therefor, an intermediate compound thereof, a pharmaceutical composition thereof and the use thereof. The PCNA inhibitor and the pharmaceutical composition comprising same can be used for treating cancers, comprising lung cancer, melanoma, colon cancer, rectal cancer, prostate cancer, ovarian cancer, leukemia, etc with PCNA overexpression.

Description

Proliferating cell nuclear antigen inhibitor, preparation method thereof, intermediate thereof, pharmaceutical composition thereof and use thereof Technical Field

[0001] The present invention relates to a proliferating cell nuclear antigen inhibitor, a preparation method thereof, a pharmaceutical composition thereof and use thereof in treating cancers such as lung cancer, melanoma, colon cancer, rectal cancer, prostate cancer, ovarian cancer and leukemia. Background Art

[0002] Proliferating cell nuclear antigen (PCNA) is a highly conserved protein that plays an important role in gene replication and repair. In cancer cells, PCNA expression is often significantly increased because they require increased DNA replication to maintain their rapid proliferation. Furthermore, PCNA is involved in DNA repair and damage response in tumor cells. PCNA can be used as both a cancer marker and an important target for cancer treatment (Oncogene 2023, 42, 613).

[0003] A Chinese patent (CN108348512) disclosed that AOH-1996 and its analogs can target and bind to PCNA, showing significant anti-cancer effects in a variety of cancer cell lines and cancer models, with minimal side effects on normal cells.

[0004] Among the compounds targeting PCNA, ATX-101 shows anti-tumor activity in a variety of tumor cells while having little effect on healthy cells. It is the first compound to enter clinical trials and show anti-tumor efficacy (Oncogene 2023,42,541).

[0005] There is little research on PCNA inhibitors, and there is a significant clinical need. Therefore, it is necessary to discover and develop safer and more effective PCNA inhibitors to improve the efficacy of cancer treatment. Summary of the Invention

[0006] The inventors have discovered that replacing the methoxybenzene ring of AOH-1996 with a bicyclic ring, or further replacing the oxygen (O) between the two benzene rings with a sulfur (S) ring, significantly impacts its antitumor activity. For example, as shown in the following structural formula, MTB-1956, which incorporates a cyclopentyl ether at the 3,4-positions of the phenyl group of AOH-1996, exhibits three times the activity of AOH-1996 in inhibiting human acute promyelocytic leukemia (HL-60) cells. Conversely, MTB-1962, which incorporates a five-membered heterocyclic ring (NS) at the 3,4-positions, exhibits a 32-fold decrease in activity. Through extensive screening, the inventors have obtained more potent PCNA inhibitors, improving cancer treatment efficacy. Furthermore, deuterating the hydrogen atoms in these inhibitors can improve their pharmacokinetic properties.

[0007] Thus, the present invention provides a new PCNA inhibitor that can be used to treat cancers such as lung cancer, melanoma, colon cancer, rectal cancer, prostate cancer, ovarian cancer, and leukemia, especially cancers in which PCNA is overexpressed.

[0008] The present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0009] in:

[0010] B is selected from substituted or unsubstituted naphthyl, substituted or unsubstituted quinolyl, or substituted or unsubstituted isoquinolyl; alternatively, B is selected from substituted or unsubstituted naphthyl; alternatively, B is naphthyl; alternatively, the substituents in the substituted naphthyl, substituted quinolyl, and substituted isoquinolyl are each independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl, halogen, or halogenated C 1-6 Alkyl; optionally, the substituents are each independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, phenyl, F, Cl, Br, I, or -CF3;

[0011] R is each independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -NH2, -CN, halogen (such as F, Cl, Br, I), -OH, or -OR', wherein R' is C 1-6 Alkyl, C 3-7 Cycloalkyl, or substituted or unsubstituted C 6-10Aryl; optionally, the alkyl is methyl, ethyl, n-propyl, or isopropyl; the cycloalkyl is cyclopropyl; the substituents of the substituted aryl and substituted heteroaryl are each independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl, halogen, or halogenated C 1-6 Alkyl; optionally, the substituents are each independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, phenyl, F, Cl, Br, I, or -CF3;

[0012] n is an integer selected from 0-3; optionally, n is 0;

[0013] Y is O or S; preferably, Y is O; preferably, Y is S;

[0014] Ar is a carbocyclic-fused phenyl, a heterocyclic-fused phenyl, a carbocyclic-fused heteroaryl, or a heterocyclic-fused heteroaryl; alternatively, Ar is a carbocyclic-fused phenyl, or a heterocyclic-fused phenyl; the carbocyclic ring is a saturated or non-aromatic unsaturated 5- or 6-membered carbocyclic ring, the heterocyclic ring is a saturated or non-aromatic unsaturated 5- or 6-membered heterocyclic ring containing 1 or 2 N atoms, or 1 or 2 O atoms, or 1 or 2 S atoms, and the heteroaryl is a 6-membered heteroaryl ring containing 1 or 2 N atoms; alternatively, Ar is a heterocyclic-fused phenyl, the heterocyclic ring is a saturated or non-aromatic unsaturated 5- or 6-membered heterocyclic ring containing 1 or 2 O atoms;

[0015] The carbocyclic fused phenyl, heterocyclic fused phenyl, carbocyclic fused heteroaryl, and heterocyclic fused heteroaryl are optionally substituted by one or more substituents, each of which is independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, OH, CN, NH2, mono- or di-C 1-6 Alkylamino, halogen, or halogenated C 1-6 alkyl; optionally, the carbocyclic-fused phenyl, heterocyclic-fused phenyl, carbocyclic-fused heteroaryl, and heterocyclic-fused heteroaryl are optionally each independently substituted by 1 or 2 substituents, each of which is independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CF3, or -CF3; optionally, the substituents are each independently selected from -CH3, -F, -Cl, or -CF3.

[0016] Alternatively, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0017] Ar is selected from the following groups:

[0018] Among them, X 1 、X 2 、X 3 、X 4 、X 5 , and X 6 Each independently selected from H, C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, or isopropyl), C 3-7 Cycloalkyl (e.g., cyclopropyl), OH, CN, NH2, halogen (e.g., Cl, Br, I, or F), or halogenated C 1-6 Alkyl; optionally, X 1 、X 4 , and X 5 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CF3, or -CF3, preferably X 1 and / or X 4 and / or X 5 is hydrogen; optionally, X 1 、X 4 , and X 5 Each independently selected from H, -CH3, -F, -Cl, or -CF3, preferably X 1 and / or X 4 and / or X 5 is hydrogen; optionally, X 2 、X 3 , and X 6 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CF3, or -CF3, preferably X 2 and / or X 3 and / or X 6 is hydrogen; optionally, X 2 、X 3 , and X 6 Each independently is H, or -CH3, preferably X 2 、X 3 , and X 6 Both of them are hydrogen or both are hydrogen.

[0019] Alternatively, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0020] Ar is selected from the following groups:

[0021] wherein X is independently selected from C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, or isopropyl), C 3-7Cycloalkyl (e.g., cyclopropyl), OH, CN, NH2, halogen (e.g., Cl, Br, I, or F), or halogenated C 1-6 Alkyl; optionally, each X is independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, F, Cl, Br, I, -CH2CH3, or -CF3; optionally, each X is independently selected from -CH3, -F, -Cl, or -CF3;

[0022] m is an integer from 0 to 3; optionally, m is 0, 1 or 2.

[0023] Alternatively, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0024] The compound of formula (I) is selected from the following compounds:

[0025] wherein X is independently selected from C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, or isopropyl), C 3-7 Cycloalkyl (e.g., cyclopropyl), -OH, -CN, -NH2, halogen (e.g., Cl, Br, I, or F), or halogenated C 1-6 Alkyl; optionally, each X is independently selected from methyl, ethyl, propyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CH3, or -CF3; optionally, each X is independently selected from -CH3, -F, -Cl, or -CF3;

[0026] m is an integer from 0 to 3; optionally, m is 0, 1 or 2.

[0027] Alternatively, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0028] The compound of formula (I) or its deuterated compound is selected from the following compounds:

[0029] Compound 44, Compound 93, Compound 99 or Compound 100 is particularly preferred.

[0030] Optionally, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, the pharmaceutically acceptable salt is a salt formed by the compound of formula (I) or its deuterated compound and an acid, and the acid includes an inorganic acid or an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or carbonic acid; optionally, the organic acid includes formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, phthalic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid.

[0031] The pharmaceutically acceptable salts can be prepared by conventional methods in the art.

[0032] The compounds represented by formula (I) of the present invention, their pharmaceutically acceptable salts, or deuterated compounds thereof include their resonance tautomers, racemates, enantiomers, diastereomers and other optical isomers.

[0033] The compound represented by formula (I) of the present invention, its pharmaceutically acceptable salt, or its deuterated compound may exist in the form of an amorphous form, a crystal, a solvent compound (such as a hydrate), or the like.

[0034] The present invention also provides a method for preparing the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, which comprises the following reaction scheme:

[0035] The definitions of Ar, Y, R, n and B in the above structural formulae are the same as those described above.

[0036] The reaction process of this reaction route is as follows: Compound Ia, which serves as an aniline precursor, and Compound Ib, which serves as a glycine derivative, undergo condensation reaction in an organic solvent (e.g., DMF) under the action of a condensation agent (polypeptide condensation reagent (HATU)) and a catalyst (e.g., N,N-diisopropylethylamine (DIPEA)) to obtain the target product, Compound I or a deuterated compound thereof.

[0037] The present invention also relates to the following intermediate compounds, among which compound 44-37 can be used to synthesize compound 44; compound 93-38 can be used to synthesize compound 93; compound 99-39 can be used to synthesize compound 99; compound 100-40 can be used to synthesize compound 100; compound 44-37-6 can be used to synthesize compound 44-37, compound 44, compound 99-39 and compound 99; compound 93-98-5 can be used to synthesize compound 93-38, compound 93, compound 100-40 and compound 100.

[0038] The present invention also provides a pharmaceutical composition comprising one or more of the compounds of formula (I) according to the present invention, or pharmaceutically acceptable salts thereof, or deuterated compounds thereof, and optionally pharmaceutically acceptable excipients. Optionally, the pharmaceutical composition further comprises an anticancer drug other than the compound of formula (I), or pharmaceutically acceptable salts thereof, or deuterated compounds thereof; preferably, the anticancer drug comprises a chemotherapeutic drug, or / and a targeted drug, or / and a nuclear drug. Preferably, the anticancer drugs include, but are not limited to, chemotherapy drugs such as cyclophosphamide, antimetabolites such as fluorouracil, gemcitabine and capecitabine, antibiotics such as doxorubicin, platinum anticancer drugs such as carboplatin, cisplatin and oxaliplatin, botanical drugs such as irinotecan, paclitaxel and taxotere, targeted drugs such as afatinib, trastuzumab, imatinib, panitumumab, ramucirumab and osimertinib, immune preparations such as pembrolizumab, nivolumab and pembrolizumab, ADC drugs such as gemtuzumab, emtansine trastuzumab, lucagon tuzumab, detrastuzumab and Enhertu, dual-antibody drugs such as PD-1 / CTLA-4 dual antibody Kaitanib and PD-(L)1 / VEGF dual antibody Ivoside, nuclear drugs such as lutetium [177Lu] texivirpitide injection (Pluvicto) and Lutathera, etc.

[0039] The present invention also provides the use of the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, or the pharmaceutical composition thereof in the preparation of a PCNA inhibitor.

[0040] The present invention also provides the use of the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing or treating cancer; optionally, the cancer includes lung cancer, melanoma, colon cancer, rectal cancer, prostate cancer, ovarian cancer, and leukemia; in particular, cancers in which PCNA is overexpressed.

[0041] The present invention also provides a method for preventing or treating cancer, comprising administering to an individual in need thereof a preventive or therapeutically effective amount of the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, or the pharmaceutical composition described above.

[0042] The compounds of formula (I) or pharmaceutically acceptable salts or deuterated compounds thereof, or pharmaceutical compositions containing the compounds of formula (I) or pharmaceutically acceptable salts or deuterated compounds thereof, described herein, can be used alone or in combination with other drugs. Appropriate combinations can produce synergistic effects, enhance the therapeutic efficacy of the aforementioned cancers, or reduce the toxic side effects of the drugs, or can also produce synergistic effects while reducing the toxic side effects. Such other drugs include, but are not limited to, chemotherapeutic drugs, targeted drugs, and nuclear drugs. Preferably, the anticancer drugs include, but are not limited to, chemotherapy drugs such as cyclophosphamide, antimetabolites such as fluorouracil, gemcitabine and capecitabine, antibiotics such as doxorubicin, platinum anticancer drugs such as carboplatin, cisplatin and oxaliplatin, botanical drugs such as irinotecan, paclitaxel and taxotere, targeted drugs such as afatinib, trastuzumab, imatinib, panitumumab, ramucirumab and osimertinib, immune preparations such as pembrolizumab, nivolumab and pembrolizumab, ADC drugs such as gemtuzumab, emtansine trastuzumab, lucagon tuzumab, detrastuzumab and Enhertu, dual-antibody drugs such as PD-1 / CTLA-4 dual antibody Kaitanib and PD-(L)1 / VEGF dual antibody Ivoside, nuclear drugs such as lutetium [177Lu] texivirpitide injection (Pluvicto) and Lutathera, etc. DETAILED DESCRIPTION

[0043] The following is a detailed description of the specific embodiments of the present invention. It should be understood that the specific embodiments described herein are only used to illustrate the present invention and are not intended to limit the present invention.

[0044] Preparation of compounds

[0045] Preparation Example 1 Preparation of Compound 8 (MTB-1956)

[0046] Compound 8-2:

[0047] Compound 8-1 (40 g, 232.32 mmol) was refluxed in SOCl2 (150 mL) for 2 h. The SOCl2 was removed by distillation, and 1,4-dioxane (20 mL) was added and distilled twice. The residue was dissolved in 1,4-dioxane (100 mL). NaOH (18.6 g, 464.64 mmol) was dissolved in water (232 mL), and glycine (17.4 g, 232.32 mmol) was added to an aqueous sodium hydroxide solution. After dissolution, the mixture was added dropwise to the reaction mixture at 0°C. The resulting reaction mixture was warmed to room temperature and reacted for 16 h. EA (200 mL) was added for extraction. The aqueous phase was adjusted to pH 2 with 5N HCl. The precipitated solid was filtered, washed with water, and dried to obtain compound 8-2 (41.8 g, 79%) as a pale yellow solid.

[0048] 1 H NMR (400MHz, DMSO) δ12.71(s,1H),8.88(t,J=6.0Hz,1H),8.36–8.28(m,1H),8.04(d,J=8.2Hz, 1H), 8.02–7.95 (m, 1H), 7.64 (dd, J=7.0, 1.3Hz, 1H), 7.61–7.51 (m, 3H), 4.00 (d, J=6.0Hz, 2H).

[0049] m / z(ESI)[M+H] + =230.1.

[0050] Compound 8-4:

[0051] Compound 8-3 (5 g, 30.49 mmol) was dissolved in THF (60 mL), NaHCO3 (5.1 g, 60.98 mmol) was added, and H2O2 (30%, 6.2 mL, 60.98 mmol) was added dropwise at 0°C. The mixture was reacted at room temperature for 16 h, and saturated sodium bisulfite solution was added to quench the reaction. The mixture was extracted with EA (200 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (methanol: CH2Cl2 = 1:30-1:20) to give compound 8-4 (4.1 g, 99%) as a white solid.

[0052] 1 H NMR (400MHz, CDCl3) δ6.76–6.69(m,1H),6.64(d,J=8.5Hz,1H),6.57(dd,J=8.5,2.6Hz,1H),4.73(s,1H),4.54(t,J=8.6Hz,2H),3.16(t,J=8.6Hz,2H).

[0053] m / z(ESI)[MH] - =135.1.

[0054] Compound 8-6:

[0055] Compound 8-5 (3.5 g, 25.09 mmol), compound 8-4 (4.1 g, 30.11 mmol), and K2CO3 (3.8 g, 27.60 mmol) were added to DMF (50 mL) under nitrogen protection and reacted at 100°C for 16 h. The mixture was cooled and extracted with EA (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:5-1:3) to obtain compound 8-6 (6.4 g, 83%) as a white solid.

[0056] m / z(ESI)[M+Na] + =279.9.

[0057] Compound 8-7:

[0058] Compound 8-6 (6.4 g, 24.9 mmol) was dissolved in a mixed solvent of EA (50 mL) and MeOH (50 mL), and 0.6 g of 10% Pd / C and hydrogen were added. The mixture was reacted at room temperature for 16 h, filtered through celite, washed with EA, and the filtrate was concentrated to obtain compound 8-7 (5.6 g, 99%) as a gray oil.

[0059] 1 H NMR (400MHz, CDCl3) δ6.98–6.89(m,1H),6.88–6.83(m,1H),6.81(dd,J=7.8,1.6Hz,1H),6.7 9–6.74(m,2H),6.74–6.68(m,2H),4.57(t,J=8.6Hz,2H),3.86(s,2H),3.18(t,J=8.6Hz,2H).

[0060] m / z(ESI)[M+H] + =228.1.

[0061] Compound 8:

[0062] Compound 8-7 (2 g, 8.8 mmol), compound 8-2 (2.4 g, 10.56 mmol), and HATU (5.0 g, 13.2 mmol) were added to DMF (25 mL), and then DIPEA (4.6 mL, 26.4 mmol) was added dropwise. Under nitrogen protection, the mixture was reacted at room temperature for 16 h. EA (100 mL) was added for extraction, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 2:10-1:2) and recrystallized (EA / petroleum ether) to obtain compound 8 (2.2 g, 57%) as an off-white solid.

[0063] 1H NMR (400MHz, CDCl3) δ8.53(s,1H),8.39(dd,J=7.9,1.1Hz,1H),8.34(d,J=8.4Hz,1H),7.9 2(d,J=8.3Hz,1H),7.89–7.82(m,1H),7.64(dd,J=7.0,1.0Hz,1H),7.48(dtd,J=16.6,6.9, 1.3Hz,2H),7.39(dd,J=8.1,7.2Hz,1H),7.02(dtd,J=24.5,7.6,1.4Hz,2H),6.90-6.80(m, 2H), 6.80-670 (m, 3H), 4.58 (t, J = 8.7Hz, 2H), 4.39 (d, J = 5.5Hz, 2H), 3.15 (t, J = 8.7Hz, 2H).

[0064] m / z(ESI)[M+H] + =439.1.

[0065] Preparation Example 2 Preparation of Compound 9 (MTB-1959)

[0066] Compound 9-9:

[0067] Compound 9-8 (5 g, 25.12 mmol), pinacol diboronate (7.7 g, 30.14 mmol), potassium acetate (7.4 g, 75.36 mmol), Pd(dppf)Cl2 (919 mg, 1.26 mmol) were added with Dioxane (50 mL) and reacted at 90 ° C for 6 h under nitrogen protection. The mixture was cooled to room temperature, EA (100 mL) and water (20 mL) were added, mixed, filtered through celite, and the filtrate was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:5-1:3) to give compound 9-9 (6 g, 97%) as a light yellow solid.

[0068] 1 H NMR (400MHz, CDCl3) δ7.32 (d, J = 7.4Hz, 1H), 7.22-7.19 (m, 2H), 4.54 (t, J = 8.7Hz, 2H), 3.21 (t, J = 8.7Hz, 2H), 1.33 (s, 12H).

[0069] Compounds 9-10:

[0070] Compound 9-9 (6 g, 24.38 mmol) was dissolved in THF (50 mL), NaHCO3 (4.1 g, 48.76 mmol) was added, and then H2O2 (30%, 5 mL, 48.76 mmol) was added dropwise at 0°C. The mixture was reacted at room temperature for 3 h, and saturated sodium bisulfite solution was added for quenching. EA (100 mL) was added for extraction, and the mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (methanol: CH2Cl2 = 1:30-1:20) to give compound 9-10 (3.23 g, 97%) as a colorless oil.

[0071] 1 H NMR (400MHz, CDCl3) δ7.08–6.88(m,1H),6.39–6.19(m,2H),4.87(s,1H),4.57(t,J=8.6Hz,2H),3.12(t,J=8.6Hz,2H).

[0072] m / z(ESI)[M+H] + =137.1.

[0073] Compounds 9-11:

[0074] Compound 8-5 (3.0 g, 21.57 mmol), compound 9-10 (3.23 g, 23.72 mmol) and K2CO3 (3.23 g, 23.72 mmol) were added to DMF (40 mL), reacted at 110°C for 16 h under nitrogen protection, cooled, extracted with EA (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to obtain compound 9-11 (5.08 g, 92%) as a yellow solid.

[0075] Compounds 9-12:

[0076] Compound 9-11 (5.08 g, 19.7 mmol) was dissolved in a mixed solvent of EA (40 mL) and MeOH (40 mL), 10% Pd / C (0.5 g) was added, hydrogen was introduced, and the reaction was carried out at room temperature for 16 h. The mixture was filtered through celite, washed with EA, and the filtrate was concentrated to obtain compound 9-12 (4.4 g, 98%) as a yellow oil.

[0077] 1H NMR (400MHz, CDCl3) δ7.07(dd,J=7.2,1.1Hz,1H),7.02–6.93(m,1H),6.88(dd,J=8.0,1.4Hz,1H),6.81(dd,J=7.9,1.5H z,1H),6.71(ddd,J=8.0,7.5,1.6Hz,1H),6.51–6.42(m,2H),4.59(t,J=8.7Hz,2H),3.78(s,2H),3.16(t,J=8.4Hz,2H).

[0078] m / z(ESI)[M+H] + =228.1.

[0079] Compound 9:

[0080] Compound 9-12 (2 g, 8.8 mmol), compound 8-2 (2.4 g, 10.56 mmol) and HATU (5.0 g, 13.2 mmol) were added to DMF (25 mL), and then DIPEA (4.6 mL, 26.4 mmol) was added dropwise to the reaction solution. The mixture was reacted at room temperature for 16 h under nitrogen protection, and EA (100 mL) was added for extraction. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and recrystallized (EA / petroleum ether) to obtain compound 9 (1.8 g, 47%) as a yellow solid.

[0081] 1 H NMR (400MHz, CDCl3) δ8.49–8.36(m,2H),8.36–8.27(m,1H),7.91(d,J=8.2Hz,1H),7.89–7.80(m,1H),7.63(d,J=6.4Hz,1H),7.57–7.44(m,2H),7 .44–7.34(m,1H),7.15-7.0(m,3H),6.89(d,J=8.1Hz,2H),6.55–6.40(m, 2H), 4.59 (t, J = 8.7Hz, 2H), 4.35 (d, J = 5.4Hz, 2H), 3.15 (t, J = 8.7Hz, 2H).

[0082] m / z(ESI)[M+H] + =439.1.

[0083] Preparation Example 3 Preparation of Compound 15 (MTB-1958)

[0084] Compound 15-14:

[0085] Compound 8-5 (5.0 g, 35.44 mmol), compound 15-13 (5.4 g, 38.98 mmol) and K2CO3 (5.4 g, 38.98 mmol) were added to DMF (50 mL), reacted at 100°C for 16 h under nitrogen protection, cooled to room temperature, added with EA (200 mL) for extraction, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to obtain compound 15-14 (9.0 g, 98%) as a yellow solid.

[0086] Compound 15-15:

[0087] The above-obtained compound 15-14 (9 g) was dissolved in a mixed solvent of EA (80 mL) and MeOH (40 mL), 10% Pd / (0.9 g) was added, hydrogen was introduced, and the reaction was carried out at room temperature for 16 h. The mixture was filtered through celite, washed with EA, and the filtrate was concentrated to obtain compound 15-15 (8 g, 98%) as a brown oil.

[0088] 1 H NMR(400MHz, CDCl3)δ6.98–6.90(m,1H),6.81(dt,J=7.8,1.5Hz,2H),6.76–6.66(m ,2H),6.56(d,J=2.4Hz,1H),6.44(dd,J=8.4,2.5Hz,1H),5.95(s,2H),3.82(s,2H).

[0089] m / z(ESI)[M+H] + =230.1.

[0090] Compound 15:

[0091] Compound 15-15 (2 g, 8.72 mmol), compound 8-2 (2.4 g, 10.46 mmol) and HATU (5.0 g, 13.08 mmol) were added to DMF (25 mL), and then DIPEA (4.6 mL, 26.16 mmol) was added dropwise. Under nitrogen protection, the mixture was reacted at room temperature for 16 h. EA (100 mL) was added for extraction, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and recrystallized (EA / petroleum ether) to obtain compound 15 (0.7 g, 18%) as an off-white solid.

[0092] 1H NMR(400MHz, CDCl3)δ8.50(s,1H),8.39(d,J=6.9Hz,1H),8.36–8.29(m,1H),7.92(d,J=8.2Hz,1H),7.8 8–7.81(m,1H),7.64(dd,J=7.0,1.0Hz,1H),7.58–7.45(m,2H),7.40(dd,J=8.1,7.2Hz,1H),7.08(td,J= 7.8,1.3Hz,1H),7.01(td,J=7.8,1.4Hz,1H),6.87(t,J=5.0Hz,1H),6.81(dd,J=8.1,1.2Hz,1H),6.73( d, J=8.4Hz, 1H), 6.56 (d, J=2.4Hz, 1H), 6.47 (dd, J=8.4, 2.4Hz, 1H), 5.96 (s, 2H), 4.38 (d, J=5.5Hz, 2H).

[0093] m / z(ESI)[M+H] + =441.2.

[0094] Preparation Example 4 Preparation of Compound 17 (MTB-1961):

[0095] Compound 17-17:

[0096] Compound 8-5 (3.0 g, 21.26 mmol), compound 17-16 (3.6 g, 23.39 mmol) and K2CO3 (3.23 g, 23.39 mmol) were added to DMF (50 mL), reacted at 100°C for 16 h under nitrogen protection, cooled to room temperature, extracted with EA (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to obtain compound 17-17 (5.8 g, 99%) as a yellow solid.

[0097] m / z(ESI)[M+H] + =274.0.

[0098] Compounds 17-18:

[0099] Compound 17-17 (5.8 g, 21.21 mmol) was dissolved in a mixed solvent of EA (50 mL) and MeOH (40 mL), 10% Pd / C (0.6 g) was added, hydrogen was introduced, and the reaction was carried out at room temperature for 16 h. The mixture was filtered through celite, washed with EA, and the filtrate was concentrated to obtain a red oil 17-18 (5.0 g, 97%).

[0100] m / z(ESI)[M+H] + =244.1.

[0101] Compound 17:

[0102] Compound 17-18 (2 g, 8.22 mmol), compound 8-2 (2.26 g, 9.86 mmol) and HATU (4.7 g, 12.33 mmol) were added to DMF (25 mL), and then DIPEA (4.3 mL, 24.66 mmol) was added dropwise. Under nitrogen protection, the mixture was reacted at room temperature for 16 h. EA (100 mL) was added for extraction, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and recrystallized (EA / petroleum ether) to obtain compound 17 (0.5 g, 13%) as a yellow solid.

[0103] 1 H NMR (400MHz, CDCl3) δ8.47(s,1H),8.38(d,J=7.5Hz,1H),8.36–8.29(m,1H),7.91(d,J=8. 2Hz,1H),7.87–7.81(m,1H),7.64(d,J=7.0Hz,1H),7.57–7.44(m,2H),7.40(t,J=7.7Hz,1H ),7.07(td,J=7.8,1.2Hz,1H),7.04–6.98(m,1H),6.90(s,1H),6.83(dd,J=11.1,4.9Hz,2 H), 6.58 (d, J = 2.8Hz, 1H), 6.52 (dd, J = 8.8, 2.8Hz, 1H), 4.36 (d, J = 5.4Hz, 2H), 4.23 (s, 4H).

[0104] m / z(ESI)[M+H] + =455.1.

[0105] Preparation Example 5 Preparation of Compound 20 (MTB-1965):

[0106] Compounds 20-21:

[0107] Compound 20-19 (5.0 g, 23.7 mmol) and compound 20-20 (7.2 g, 28.4 mmol) were placed in a round-bottom flask, potassium acetate (6.98 g, 71.1 mmol) and PdCl2(dppf) (0.87 g, 1.19 mmol) were added, and the mixture was dissolved in 1,4-dioxane (60 mL). The reaction was stirred at 90°C for 3 h. The mixture was quenched with water at 0°C, extracted with EA (100 mL), filtered through celite, washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:3) to afford compound 20-21 (2.36 g, 38%) as a yellow solid.

[0108] 1 H NMR (400MHz, CDCl3) δ7.91 (s, 2H), 7.82 (d, J = 8.2Hz, 1H), 7.75 (d, J = 8.2Hz, 1H), 3.87 (s, 3H), 1.39 (s, 12H).

[0109] m / z(ESI)[M+H] + =259.

[0110] Compounds 20-22:

[0111] Compound 20-21 (2.36 g, 9.14 mmol) was placed in a round-bottom flask and dissolved in THF (25 mL). Sodium bicarbonate (1.54 g, 18.29 mmol) was added, and hydrogen peroxide (30%, 1.87 mL, 18.3 mmol) was added dropwise at 0°C. The reaction was stirred at room temperature for 3 h. The mixture was quenched with saturated sodium bisulfite, concentrated, and co-evaporated twice with methanol. The mixture was purified by silica gel column chromatography (methanol:CH2Cl2 = 1:30-1:20) to afford compound 20-22 (1.0 g) as a white solid, which was used directly in the next step.

[0112] 1 H NMR (400MHz, DMSO) δ9.27 (s, 1H), 7.94 (s, 1H), 7.40 (d, J = 8.6Hz, 1H), 6.82 (d, J = 2.2Hz, 1H), 6.69 (dd, J = 2.2, 8.6Hz, 1H), 3.72 (s, 3H).

[0113] m / z(ESI)[M+H] + =149.

[0114] Compounds 20-23:

[0115] Compound 20-22 (1.0 g, 6.82 mmol) and compound 8-5 (875 mg, 6.2 mmol) were placed in a round-bottom flask, potassium carbonate (943 mg, 6.82 mmol) was added, and the mixture was dissolved in DMF (20 mL). The mixture was reacted at 100°C for 16 h, cooled to room temperature, quenched with water at 0°C, and extracted with EA (100 mL). The EA layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:3) to obtain compound 20-23 (1.7 g, 92%) as a yellow solid.

[0116] 1 H NMR (400MHz, CDCl3) δ7.98(dd,J=1.6,8.0Hz,1H),7.89(s,1H),7.79(d,J=8.7Hz,1H),7.51-7.46(m,1H),7.19(td ,J=1.2,7.7Hz,1H),7.13(d,J=2.3Hz,1H),7.05(dd,J=2.3,8.7Hz,1H),6.96(dd,J=1.0,8.4Hz,1H),3.83(s,3H).

[0117] m / z(ESI)[M+H] + =270.

[0118] Compounds 20-24:

[0119] Compound 20-23 (1.7 g, 6.31 mmol) was placed in a round-bottom flask, and EA (15 mL) and MeOH (10 mL) were added, followed by 10% Pd / C (0.2 g). Hydrogen was introduced and the mixture was reacted at room temperature for 16 h. The mixture was filtered through celite, concentrated, and purified by silica gel column chromatography (methanol:CH2Cl2=1:30-1:10) to give compound 20-24 (1.5 g, 99%) as a foamy solid.

[0120] 1 H NMR (400MHz, CDCl3) δ7.83 (s, 1H), 7.75 (d, J = 8.7Hz, 1H), 7.06 (dd, J = 2.3, 8.8Hz, 1H), 7.03-6 .99(m,1H),6.94(d,J=2.2Hz,1H),6.87(d,J=7.8Hz,2H),6.74(td,J=1.6,7.8Hz,1H),3.88(br s,2H),3.77(s,3H).

[0121] m / z(ESI)[M+H] + =240.

[0122] Compound 20:

[0123] Compound 20-24 (1.55 g, 6.48 mmol) and compound 8-2 (1.78 g, 7.77 mmol) were placed in a round-bottom flask, HATU (3.7 g, 9.72 mmol) was added, and the mixture was dissolved in DMF (20 mL). Finally, DIPEA (3.4 mL, 19.44 mmol) was added and allowed to react at room temperature for 16 h. The mixture was quenched with water at 0°C and extracted with EA (100 mL). The EA layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:2) to obtain a solid. The solid was recrystallized from petroleum ether and ethyl acetate to afford compound 20 (0.9 g, 31%) as a white solid.

[0124] 1 H NMR (400MHz, DMSO) δ9.65(s,1H),8.95(d,J=6.0Hz,1H),8.33(d,J=8.0Hz,1H),8.20(d,J= 7.8Hz,1H),8.18(s,1H),8.02(d,J=8.2Hz,1H),7.97(d,J=7.6Hz,1H),7.67(d,J=8.6Hz,1 H),7.65(d,J=7.2Hz,1H),7.57-7.46(m,3H),7.31(d,J=2.2Hz,1H),7.13-7.04(m,2H),6. 89 (dd, J = 2.2, 8.6 Hz, 1H), 6.81 (dd, J = 1.4, 7.8 Hz, 1H), 4.19 (d, J = 6.0 Hz, 2H), 3.77 (s, 3H).

[0125] m / z(ESI)[M+H] + =451.

[0126] Preparation Example 6 Preparation of Compound 21 (MTB-1963):

[0127] Compounds 21-26:

[0128] Compound 21-25 (5.0 g, 33.75 mmol) and compound 8-5 (4.33 g, 30.68 mmol) were placed in a round-bottom flask, potassium carbonate (4.66 g, 33.75 mmol) was added, and the mixture was dissolved in DMF (80 mL). The reaction was allowed to proceed at 100°C overnight, cooled to room temperature, quenched with water at 0°C, and extracted with EA (100 mL). The EA layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:2) to obtain compound 21-26 (5.2 g, 57%) as a yellow-white solid.

[0129] 1 H NMR (400MHz, CDCl3) δ7.97(dd,J=1.6,8.0Hz,1H),7.90(s,1H),7.50(d,J=2.2Hz,1H),7.48-7.41(m,1H),7. 40(d,J=8.6Hz,1H),7.18(d,J=7.6Hz,1H),7.13(dd,J=1.8,8.7Hz,1H),6.96(d,J=8.4Hz,1H),3.90(s,3H).

[0130] m / z(ESI)[M+H] + =270.

[0131] Compounds 21-27:

[0132] Compound 21-26 (5.2 g, 19.31 mmol) was placed in a round-bottom flask, and EA (45 mL) and MeOH (30 mL) were added, followed by 10% Pd / C (0.2 g). Hydrogen was introduced and the mixture was reacted at room temperature for 16 h. The mixture was filtered through celite, concentrated, and dried in vacuo to obtain a foamy solid compound 21-27 (4.6 g, 99%).

[0133] 1 H NMR (400MHz, CDCl3) δ7.87(s,1H),7.42(d,J=2.2Hz,1H),7.34(d,J=8.6Hz,1H),7.10(dd,J= 2.2,8.6Hz,1H),6.97(td,J=1.4,7.6Hz,1H),6.86-6.81(m,2H),6.68-6.72(m,1H),3.90(br s,2H),3.86(s,3H).

[0134] m / z(ESI)[M+H] + =240.

[0135] Compound 21:

[0136] Compound 21-27 (2.52 g, 10.53 mmol) and compound 8-2 (2.9 g, 12.64 mmol) were placed in a round-bottom flask, HATU (6.0 g, 15.8 mmol) was added, and the mixture was dissolved in DMF (30 mL). Finally, DIPEA (5.5 mL, 31.59 mmol) was added and allowed to react at room temperature for 16 h. The mixture was quenched with water at 0°C and extracted with EA (100 mL). The EA layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:2) to obtain a solid. The solid was recrystallized from ethyl acetate to give compound 21 (180 mg, 21%) as a white solid.

[0137] 1 H NMR (400MHz, DMSO) δ9.65(s,1H),8.94(t,J=6.0Hz,1H),8.34(d,J=8.0Hz,1H),8.22(s,1H),8.20(d,J=7.8Hz,1H),8.02(d,J=8.2Hz,1H),7. 98(d,J=7.6Hz,1H),7.47-7.65(m,5H),7.32(d,J=2.2Hz,1H),7.12-7.02(m,3H),6.75(d,J=7.8Hz,1H),4.21(d,J=6.0Hz,2H),3.85(s,3H).

[0138] m / z(ESI)[M+H] + =451.

[0139] Preparation Example 7 Preparation of Compound 23 (MTB-1957)

[0140] Compounds 23-29:

[0141] Compound 23-28 (5 g, 23.8 mmol, pinacol diboronate (9.1 g, 35.7 mmol), potassium acetate (7.0 g, 71.4 mmol) and Pd(dppf)Cl2 (871 mg, 1.19 mmol) were added to Dioxane (60 mL), and the mixture was reacted at 100°C for 6 h under nitrogen protection. The mixture was cooled to room temperature, and EA (100 mL) and water (20 mL) were added. The mixture was filtered through celite and the filtrate was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to give compound 23-29 (5.9 g, 96%) as a white solid.

[0142] 1H NMR (400MHz, CDCl3) δ8.16(s,1H),7.67(d,J=8.3Hz,1H),7.32(d,J=8.3Hz,1 H), 7.04 (d, J = 3.1Hz, 1H), 6.50 (d, J = 3.1Hz, 1H), 3.79 (s, 3H), 1.37 (s, 12H).

[0143] Compounds 23-30:

[0144] Compound 23-29 (5.9 g, 22.94 mmol) was dissolved in THF (50 mL), and NaHCO3 (3.8 g, 45.88 mmol) was added. H2O2 (30%, 4.7 mL, 45.88 mmol) was then added dropwise at 0°C. The mixture was reacted at room temperature for 3 h, and saturated sodium bisulfite solution was added for quenching. EA (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (methanol: CH2Cl2 = 1:30-1:20) to give compound 23-30 (1.38 g, 41%) as a yellow solid.

[0145] 1 H NMR (400MHz, CDCl3) δ7.17(d,J=8.7Hz,1H),7.08–6.94(m,2H),6.81(dd,J=8.7,2.4Hz,1H),6.35(dd,J=3.0,0.7Hz,1H),5.01(s,1H),3.75(s,3H).

[0146] m / z(ESI)[M+H] + =148.1.

[0147] Compound 23-31:

[0148] Compound 8-5 (1.2 g, 8.52 mmol), compound 23-30 (1.38 g, 9.37 mmol) and K2CO3 (1.3 g, 9.37 mmol) were added to DMF (30 mL), reacted at 110°C for 16 h under nitrogen protection, cooled to room temperature, and extracted with EA (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to obtain compound 23-31 (2.08 g, 91%) as a yellow solid.

[0149] m / z(ESI)[M+H] + =269.1.

[0150] Compound 23-32:

[0151] The above-obtained compound 23-31 (2.08 g, 7.7 mmol) was dissolved in a mixed solvent of EA (30 mL) and MeOH (30 mL), 10% Pd / C (0.2 g) was added, hydrogen was introduced, and the reaction was carried out at room temperature for 16 h. The mixture was filtered through celite, washed with EA, and the filtrate was concentrated to obtain compound 23-32 (1.8 g, 98%) as a gray oil.

[0152] m / z(ESI)[M+H] + =239.1.

[0153] Compound 23:

[0154] The above-obtained compound 23-32 (1.8 g, 7.55 mmol), compound 8-2 (2.1 g, 9.06 mmol) and HATU (4.3 g, 11.33 mmol) were added to DMF (25 mL), and then DIPEA (3.9 mL, 22.65 mmol) was added dropwise. Under nitrogen protection, the mixture was reacted at room temperature for 16 h. EA (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and recrystallized (EA / petroleum ether) to obtain compound 23 (1.7 g, 50%) as a white solid.

[0155] 1 H NMR (400MHz, CDCl3) δ8.54(s,1H),8.42(d,J=7.1Hz,1H),8.35(d,J=8.3Hz,1H),7.90(d,J=8.3 Hz,1H),7.86–7.81(m,1H),7.64(dd,J=7.0,1.0Hz,1H),7.54–7.41(m,2H),7.40–7.32(m,1H), 7.30–7.24(m,2H),7.10(d,J=3.1Hz,1H),7.05(td,J=7.9,1.3Hz,1H),7.02–6.93(m,2H),6.83 (s, 1H), 6.77 (dd, J = 8.1, 1.1Hz, 1H), 6.43 (d, J = 3.0Hz, 1H), 4.39 (d, J = 5.4Hz, 2H), 3.80 (s, 3H).

[0156] m / z(ESI)[M+H] + =450.1.

[0157] Preparation Example 8 Preparation of Compound 24 (MTB-1960):

[0158] Compounds 24-34:

[0159] Compound 24-33 (3.4 g, 13.22 mmol) was dissolved in THF (50 mL), and NaHCO3 (2.2 g, 26.44 mmol) was added. H2O2 (30%, 2.7 mL, 26.44 mmol) was then added dropwise at 0°C. The mixture was reacted at room temperature for 3 h, and saturated sodium bisulfite solution was added for quenching. EA (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (methanol: CH2Cl2 = 1:30-1:20) to give compound 24-34 (1.9 g, 97%) as a yellow oil.

[0160] 1 H NMR (400MHz, CDCl3) δ7.45(d,J=8.4Hz,1H),6.93(d,J=3.1Hz,1H),6.77(d,J=2.2Hz,1 H), 6.67 (dd, J=8.4, 2.2Hz, 1H), 6.41 (dd, J=3.1, 0.7Hz, 1H), 4.82 (s, 1H), 3.70 (s, 3H).

[0161] m / z(ESI)[M+H] + =148.1.

[0162] Compounds 24-35:

[0163] Compound 8-5 (1.5 g, 10.76 mmol), compound 24-34 (1.9 g, 12.91 mmol) and K2CO3 (1.6 g, 11.8 mmol) were added to DMF (30 mL), reacted at 110°C under nitrogen protection for 16 h, cooled to room temperature, added with EA (100 mL) for extraction, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to obtain a yellow solid 24-35 (2.4 g, 83%).

[0164] m / z(ESI)[M+H] + =269.1.

[0165] Compounds 24-36:

[0166] The above-obtained compound 24-35 (2.4 g, 8.9 mmol) was dissolved in a mixed solvent of EA (30 mL) and MeOH (30 mL), 10% Pd / C (0.2 g) was added, hydrogen was introduced, and the reaction was carried out at room temperature for 16 h. The mixture was filtered, washed with EA, and the filtrate was concentrated to obtain a gray oil 24-36 (2.1 g, 99%).

[0167] m / z(ESI)[M+H] + =239.1.

[0168] Compound 24:

[0169] Compound 24-36 (2.1 g, 8.81 mmol), compound 8-2 (2.4 g, 10.57 mmol) and HATU (5.0 g, 13.22 mmol) were added to DMF (25 mL), and then DIPEA (4.6 mL, 26.43 mmol) was added dropwise. Under nitrogen protection, the mixture was reacted at room temperature for 16 h. EA (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and recrystallized (EA / petroleum ether) to obtain compound 24 (154 mg, 39%) as a white solid.

[0170] 1 H NMR(400MHz, DMSO)δ9.66(s,1H),8.98(t,J=5.9Hz,1H),8.39–8.32(m,1H),8.23(d,J= 7.6Hz,1H),8.06–7.91(m,2H),7.66(dd,J=7.0,0.9Hz,1H),7.62–7.43(m,4H),7.32(d ,J=3.1Hz,1H),7.20(d,J=1.9Hz,1H),7.14–6.98(m,2H),6.83(dd,J=8.5,2.1Hz,1H), 6.78(dd,J=8.0,1.5Hz,1H), 6.46(d,J=3.1Hz,1H), 4.24(d,J=6.0Hz,2H), 3.72(s,3H).

[0171] m / z(ESI)[M+H] + =450.1.

[0172] Preparation Example 9 Preparation of Compound 44

[0173] Compound 44-37 (764 mg, 3.17 mmol), compound 8-2 (872 mg, 3.80 mmol), and HATU (1.8 g, 4.76 mmol) were added to DMF (15 mL), and then DIPEA (1.7 mL, 9.51 mmol) was added dropwise. Under nitrogen protection, the mixture was reacted at room temperature for 16 h, and EA (100 mL) was added for extraction. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2), and then recrystallized (EA / petroleum ether) to obtain a yellow solid 44 (440 mg, 31%).

[0174] 1 H NMR (400MHz, CDCl3) δ8.68(s,1H),8.38(d,J=7.5Hz,1H),8.33(d,J=8.4Hz,1H),7.90 (d,J=8.1Hz,1H),7.83(d,J=8.0Hz,1H),7.64(d,J=6.8Hz,1H),7.48(t,J=7.3Hz,1H) ,7.42-7.31(m,2H),7.05-6.89(m,3H),6.79(s,1H),6.65(s,1H),6.55(d,J=7.7Hz,1 H), 4.57 (t, J = 8.5Hz, 2H), 4.41 (d, J = 5.0Hz, 2H), 3.13 (t, J = 8.3Hz, 2H), 2.07 (s, 3H). m / z(ESI)[M+H] + =453.1.

[0175] Preparation Example 10 Preparation of Compound 90:

[0176] Compound 90 was prepared using the method for preparing compound 44.

[0177] m / z(ESI)[M+H] + =469.1.

[0178] Preparation Example 11 Preparation of Compound 93 (MTB-1993):

[0179] Compound 93 was prepared from compound 93-38 using the method for preparing compound 44.

[0180] 1H NMR (400MHz, CDCl3) δ8.61(s,1H),8.40(d,J=7.5Hz,1H),8.35(d,J=8.5Hz,1H),7.92(d,J=8.2Hz,1H ),7.85(d,J=8.1Hz,1H),7.67(d,J=6.8Hz,1H),7.49(t,J=7.2Hz,1H),7.40(dd,J=16.1,8.6Hz,2H), 7.01(t,J=7.2Hz,1H),6.95(t,J=7.2Hz,1H),6.87(t,J=5.1Hz,1H),6.80(s,1H),6.66(s,1H),6.56( d,J=7.7Hz,1H),4.58(t,J=8.6Hz,2H),4.43(d,J=5.4Hz,2H),3.15(t,J=8.6Hz,2H).m / z(ESI)[M+H] + =456.3.

[0181] m / z(ESI)[M+H] + =457.2.

[0182] Preparation Example 12 Preparation of Compound 99

[0183] Compound 99-39 (500 mg, 1.93 mmol), 8-2 (575 mg, 2.51 mmol), and HATU (1.09 g, 2.90 mmol) were added to DMF (10 mL), and then DIPEA (0.98 mL, 5.79 mmol) was added dropwise. Under nitrogen protection, the reaction was allowed to react at room temperature overnight. Ethyl acetate was added for extraction, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and then recrystallized (EA / petroleum ether) to give an off-white solid 99 (408 mg, 45%). 1H NMR (400MHz, CDCl3) δ8.56 (s, 1H), 8.36-8.30 (m, 2H), 7.92 (d, J = 8.3Hz, 1H), 7.85 (d, J = 8.2Hz, 1H),7.67(dd,J=7.0,1.0Hz,1H),7.49(td,J=7.0,1.0Hz,1H),7.43–7.34(m,2H),6.86(t,J=5. 0Hz,1H),6.82(s,1H),6.70(td,J=8.5,2.7Hz,1H),6.67(s,1H),6.28(dd,J=9.8,2.7Hz,1H),4 .60(t,J=8.7Hz,2H),4.43(d,J=5.5Hz,2H),3.17(t,J=8.6Hz,2H),2.07(s,3H).m / z(ESI)[M+H] + =471.3.

[0184] Preparation Example 13 Preparation of Compound 100 (MTB-2006)

[0185] Compound 100-40 (675 mg, 2.57 mmol), 8-2 (766 mg, 3.34 mmol), and HATU (1.46 g, 3.86 mmol) were added to DMF (10 mL), and then DIPEA (1.3 mL, 7.71 mmol) was added dropwise. Under nitrogen protection, the reaction was allowed to react at room temperature overnight. Ethyl acetate was added for extraction, and the mixture was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2) and recrystallized (EA / petroleum ether) to give an off-white solid compound 100 (450 mg, 37%).

[0186] 1 H NMR (400MHz, CDCl3) δ8.59(s,1H),8.39–8.25(m,2H),7.91(d,J=8.3Hz,1H),7.85(d,J =8.2Hz,1H),7.66(dd,J=7.1,1.0Hz,1H),7.52–7.45(m,1H),7.42–7.33(m,2H),6.91(t ,J=5.2Hz,1H),6.81(s,1H),6.72–6.67(m,1H),6.66(s,1H),6.27(dd,J=9.8,2.7Hz,1 H),4.59(t,J=8.7Hz,2H),4.42(d,J=5.5Hz,2H),3.16(t,J=8.7Hz,2H).m / z(ESI)[M+H] + =474.4.

[0187] Preparation Example 14 Preparation of other analogs

[0188] Using a similar method, compounds Ia and Ib were condensed to prepare the target products shown in Table 1.

[0189] Table 1

[0190] Preparation Example 15 Preparation of Comparative Compound 1 (MTB-1962)

[0191] Compound 1-39:

[0192] Compound 1-38 (15.0 g, 70.1 mmol) and compound 20-20 (21.35 g, 84.1 mmol) were placed in a round-bottom flask, potassium acetate (20.63 g, 210.2 mmol) and PdCl2(dppf) (2.56 g, 3.5 mmol) were added, and the mixture was dissolved in 1,4-dioxane (140 mL). The reaction was stirred at 90°C for 5 h, quenched with water at 0°C, extracted with EA (100 mL), filtered through celite, and the organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:3) to obtain compound 1-39 (18.3 g, 99%) as a yellow-white solid.

[0193] m / z(ESI)[M+H] + =262.

[0194] Compound 1-40:

[0195] Compound 1-39 (6.0 g, 22.98 mmol) was placed in a round-bottom flask and dissolved in THF (50 mL). Sodium bicarbonate (3.86 g, 45.96 mmol) was added, and hydrogen peroxide (30%, 4.7 mL, 45.96 mmol) was added dropwise at 0°C. The reaction was stirred at room temperature for 3 h. The mixture was quenched with saturated sodium bisulfite and extracted with EA (100 mL). The EA layer was washed once with water and once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and recrystallized from petroleum ether and ethyl acetate to afford 2.66 g of compound 1-40 as a yellow-white solid, which was used directly in the next step.

[0196] 1H NMR (400MHz, DMSO) δ9.73 (s, 1H), 9.30 (s, 1H), 7.92 (d, J = 8.6Hz, 1H), 7.40 (d, J = 2.3Hz, 1H), 6.95 (dd, J = 2.3, 8.6Hz, 1H).

[0197] m / z(ESI)[M+H] + =152.

[0198] Compound 1-41:

[0199] Compound 1-40 (2.66 g, 17.6 mmol) and compound 8-5 (2.26 g, 16.0 mmol) were placed in a round-bottom flask, potassium carbonate (2.43 g, 17.6 mmol) was added, and the mixture was dissolved in DMF (40 mL). The mixture was reacted at 100°C for 16 h, cooled to room temperature, quenched with water at 0°C, and extracted with EA (100 mL). The EA layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:3) to obtain compound 1-41 (4.43 g, 92%) as a yellow oil.

[0200] 1 H NMR(400MHz, CDCl3)δ9.07(s,1H),8.03(dd,J=1.7,8.2Hz,1H),7.97(d,J=8.6Hz,1H),7 .74(d,J=2.3Hz,1H),7.58-7.54(m,1H),7.30-7.26(m,2H),7.12(dd,J=1.2,8.3Hz,1H).

[0201] m / z(ESI)[M+H] + =273.

[0202] Compound 1-42:

[0203] Compound 1-41 (4.43 g, 16.3 mmol) was placed in a round-bottom flask, EA (30 mL) and MeOH (20 mL) were added, and then 10% Pd / C (0.2 g) was added. Hydrogen was introduced and the reaction was carried out at room temperature for 16 h. The mixture was filtered through celite, concentrated, and dried in vacuo to obtain compound 1-42 (4.28 g) as a yellow oil, which was used directly in the next step.

[0204] 1H NMR (400MHz, CDCl3) δ9.01(s,1H),7.89(d,J=8.7Hz,1H),7.68(d,J=2.3Hz,1H),7.23(dd,J=2.3,8.7Hz,1H),7.05(t d,J=1.3,7.8Hz,1H),6.96(dd,J=1.3,7.8Hz,1H),6.87(dd,J=1.5,7.9Hz,1H),6.76(td,J=1.5,7.8Hz,1H),3.84(br s,2H).

[0205] m / z(ESI)[M+H] + =243.

[0206] Preparation of compound 44-37

[0207] Compound 44-37-2

[0208] Compound 44-37-1 (10 g, 53.46 mmol), 1,2-dibromoethane (30.1 g, 160.38 mmol), and potassium carbonate (11.1 g, 80.19 mmol) were added to acetonitrile (150 mL) and reacted at 85° C. under nitrogen protection overnight. The mixture was cooled to room temperature, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to obtain 44-37-2 (7.4 g, 47%) as a colorless oil.

[0209] 1 H NMR (400MHz, CDCl3) δ7.40 (d, J = 7.9 Hz, 1H), 6.72 (d, J = 1.5 Hz, 1H), 6.69 (dd, J = 7.9, 1.5 Hz, 1H), 4.32 (t, J = 6.6 Hz, 2H), 3.67 (t, J = 6.6 Hz, 2H), 2.31 (s, 3H).

[0210] Compound 44-37-1 can also be reacted with 2-bromoethanol in the presence of potassium carbonate and then brominated to give compound 44-37-2.

[0211] Compound 44-37-3

[0212] Compound 44-37-2 (7.4 g, 25.17 mmol) was dissolved in THF (50 mL) and n-BuLi (13.1 mL, 32.72 mmol, 2.5 M) was added dropwise at -78 ° C under nitrogen protection. The mixture was reacted at -78 ° C for 2 h, quenched with water, extracted with EA, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to give 44-37-3 (3.06 g, 90%) as a light yellow oil.

[0213] Compound 44-37-4

[0214] Compound 44-37-3 (3.06 g, 22.81 mmol) was dissolved in acetonitrile (45 mL), and NBS (4.46 g, 25.08 mmol) was added in portions at 0°C, followed by reaction at room temperature for 16 h. The solvent was distilled off, and EA was added. The organic phase was washed with saturated aqueous sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a light yellow solid 44-37-4 (4.8 g, 99%).

[0215] 1 H NMR (400MHz, CDCl3) δ7.31 (s, 1H), 6.67 (s, 1H), 4.55 (t, J = 8.7Hz, 2H), 3.16 (t, J = 8.7Hz, 2H), 2.32 (s, 3H).

[0216] Compound 44-37-5

[0217] Compound 44-37-4 (4.8 g, 22.53 mmol), pinacol diboronate (6.9 g, 27.03 mmol), potassium acetate (6.6 g, 67.59 mmol), and Pd(dppf)Cl2 (824 mg, 1.13 mmol) were added to 1,4-dioxane (50 mL) and reacted at 90°C for 6 h under nitrogen protection. The mixture was cooled to room temperature, and EA and water were added. The mixture was filtered through celite, and the filtrate was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to give 44-37-5 (2.7 g, 46%) as a light yellow solid.

[0218] Compound 44-37-6

[0219] Compound 44-37-5 (2.7 g, 10.38 mmol) was dissolved in THF (20 mL), and NaHCO3 (1.7 g, 20.76 mmol) was added. Then, 30% H2O2 (2.1 mL, 20.76 mmol) was added dropwise at 0°C. The mixture was reacted at room temperature for 3 h. Saturated sodium bisulfite solution was added to quench the mixture, and EA was added to extract the mixture. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (methanol:CH2Cl2=1:30-1:20) to give 44-37-6 (600 mg, 38%) as a colorless oil.

[0220] 1 H NMR (400MHz, CDCl3) δ6.66 (s, 1H), 6.56 (s, 1H), 4.51 (t, J = 8.7Hz, 2H), 4.33 (s, 1H), 3.13 (t, J = 8.7Hz, 2H), 2.19 (s, 3H).

[0221] Compound 44-37-7

[0222] Compound 44-37-6 (600 mg, 4.0 mmol), 8-5 (470 mg, 3.33 mmol), and K2CO3 (506 mg, 3.66 mmol) were added to DMF (10 mL) and reacted at 100°C for 16 h under nitrogen protection. The mixture was cooled to room temperature and extracted with EA. The organic phase was washed with 10% NaOH solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give 44-37-7 (860 mg, 79%) as a brown oil.

[0223] Compound 44-37

[0224] Compound 44-37-7 (860 mg, 3.17 mmol) was dissolved in a mixed solvent of ethyl acetate (10 mL) and MeOH (10 mL). 10% Pd / C (100 mg) was added and the mixture was reacted at room temperature under hydrogen for 16 h. The mixture was filtered through celite, washed with EA, and the filtrate was concentrated to give a brown oil 44-37 (764 mg, 99%). m / z (ESI) [M+H] + =242.1.

[0225] Preparation of compound 93-38

[0226] Compound 93-38-2

[0227] Compound 93-38-1 (10 g, 50.24 mmol) was dissolved in THF (100 mL). Under nitrogen protection, n-BuLi (22 mL, 55.26 mmol, 2.5 M) was added dropwise at -78 ° C. and stirred at -78 ° C for 30 min. CD3I (8.7 g, 60.29 mmol) was dissolved in THF (20 mL) and added dropwise to the reaction system. The mixture was reacted at -78 ° C for 1 h, then warmed to room temperature for 1 h. Saturated ammonium chloride solution was added to quench the reaction, and EA was added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 93-38-2 (6.3 g, 91%) as a light yellow oil.

[0228] 1 H NMR (400MHz, CDCl3) δ7.07 (d, J = 7.4 Hz, 1H), 6.66 (dd, J = 7.4, 1.4 Hz, 1H), 6.63 (d, J = 1.4 Hz, 1H), 4.55 (t, J = 8.7 Hz, 2H), 3.16 (t, J = 8.7 Hz, 2H).

[0229] Compound 93-38-3

[0230] Compound 93-38-2 (6.3 g, 45.92 mmol) was placed in a round-bottom flask, acetonitrile (90 mL) was added, and NBS (9.0 g, 50.51 mmol) was added portionwise. The mixture was allowed to react at room temperature for 16 h. The solvent was evaporated, and the mixture was extracted with EA and water. The organic layer was washed with saturated sodium bicarbonate and then saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford 93-38-3 (9.9 g, 99%) as a yellow solid.

[0231] 1 H NMR (400MHz, CDCl3) δ7.31 (s, 1H), 6.67 (s, 1H), 4.56 (t, J = 8.7Hz, 2H), 3.17 (t, J = 8.7Hz, 2H).

[0232] Compound 93-38-4

[0233] To compound 93-38-3 (9.9 g, 45.81 mmol), pinacol diboronate (17.4 g, 68.72 mmol), potassium acetate (13.5 g, 137.43 mmol), and Pd(dppf)Cl2 (1.7 g, 29 mmol) was added 1,4-dioxane (100 mL). Under nitrogen protection, the reaction was carried out at 100°C for 16 h. The mixture was cooled to room temperature, EA and water were added, and the mixture was filtered through celite. The filtrate was extracted, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to give a light yellow oil 93-38-4 (7.7 g, 64%).

[0234] Compound 93-38-5

[0235] Compound 93-38-4 (7.7 g, 29.26 mmol) was dissolved in THF (70 mL), and NaHCO3 (4.9 g, 58.52 mmol) was added. H2O2 (6 mL, 58.52 mmol, 30%) was then added dropwise at 0°C. The mixture was reacted at room temperature for 16 h, and saturated sodium bisulfite solution was added to quench the reaction. EA was added to extract the mixture, and the mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (methanol: CH2Cl2 = 1:30-1:20) to give 93-38-5 (2.4 g, 54%) as a light yellow solid.

[0236] 1 H NMR (400MHz, CDCl3) δ6.66 (s, 1H), 6.56 (s, 1H), 4.51 (t, J = 8.6 Hz, 2H), 4.34 (s, 1H), 3.13 (t, J = 8.6 Hz, 2H).

[0237] Compound 93-38-6

[0238] Compound 93-38-5 (2.4 g, 15.67 mmol), 8-5 (2.0 g, 14.24 mmol), and K2CO3 (2.2 g, 15.67 mmol) were added to DMF (30 mL) and reacted at 100°C for 16 h under nitrogen protection. The mixture was cooled to room temperature and extracted with EA. The organic phase was washed with 10% NaOH solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid 93-38-6 (1.9 g, 44%).

[0239] 1H NMR(400MHz, CDCl3)δ7.91(dd,J=8.1,1.7Hz,1H),7.43–7.37(m,1H),7.09–7.04(m,1H),6.85( s, 1H), 6.75 (dd, J = 8.5, 1.0Hz, 1H), 6.68 (s, 1H), 4.59 (t, J = 8.6Hz, 2H), 3.18 (t, J = 8.6Hz, 2H).

[0240] Compound 93-38

[0241] To compound 93-38-6 (1.7 g, 6.20 mmol) and NH4Cl (2.0 g, 37.2 mmol) was added EtOH (40 mL), followed by the addition of zinc powder (2.4 g, 37.2 mmol) in portions. The mixture was reacted at 80°C under nitrogen for 16 h, filtered, and the filtrate concentrated. The residue was purified by silica gel column chromatography (methanol:CH2Cl2 = 1:30-1:10) to afford 93-38 (1.51 g, 99%) as a red oil. m / z (ESI) [M+H] + =245.1.

[0242] Preparation of compound 99-39

[0243] Compound 99-39-2

[0244] Compounds 44-37-6 (500 mg, 3.33 mmol) and 99-39-1 (525 mg, 3.33 mmol) were dissolved in toluene (20 mL). t-BuOK (385 mg, 3.43 mmol) was added portionwise at 0°C. The mixture was allowed to react at room temperature for 2 h, quenched with water, and extracted with EA. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:3) to obtain 99-39-2 (320 mg, 33%) as a yellow solid. m / z (ESI) [M+H] + =290.1.

[0245] Compound 99-39

[0246] Compound 99-39-2 (500 mg, 1.73 mmol) and NH4Cl (555 mg, 10.38 mmol) were added to THF (20 mL) and water (10 mL), followed by the addition of zinc powder (673 mg, 10.38 mmol) in portions. The mixture was reacted at 70°C for 5 h under nitrogen protection, filtered, and the filtrate was extracted with EA. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford 99-39 (325 mg, 72%) as a brown oil. m / z (ESI) [M+H] + =260.1.

[0247] Preparation of Compound 100-40

[0248] Compound 100-40-1

[0249] Compound 93-38-5 (1.0 g, 6.53 mmol) and compound 99-39-1 (1.0 g, 6.53 mmol) were dissolved in toluene (20 mL), t-BuOK (769 mg, 6.86 mmol) was added in batches at 0°C, and then reacted at room temperature for 2 h. The mixture was quenched with water and extracted with EA. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:3) to give a yellow solid 100-40-1 (750 mg, 39%) (determined according to the literature for the ortho-selective substitution reaction of the nitro group).

[0250] 1 H NMR (400MHz, CDCl3) δ8.02 (dd, J=9.1, 5.8Hz, 1H), 6.87 (s, 1H), 6.78–6.73 (m, 1H), 6. 69 (s, 1H), 6.43 (dd, J = 10.1, 2.6Hz, 1H), 4.60 (t, J = 8.7Hz, 2H), 3.20 (t, J = 8.7Hz, 2H).

[0251] 13 C NMR (101MHz, CDCl3) δ165.61(d,J=256.6Hz),158.17,154.77(d,J=10.8Hz),145.11,135.86,129.89,128.18(d,J =11.1Hz),126.43,117.82,111.72,108.54(d,J=23.9Hz),104.20(d,J=27.3Hz),71.77,29.70,15.57–14.99(m).

[0252] Compound 100-40

[0253] Compound 100-40-1 (750 mg, 2.57 mmol) and NH4Cl (825 mg, 15.42 mmol) were added to THF (20 mL) and water (10 mL), followed by the addition of zinc powder (1.0 g, 15.42 mmol) in portions. The mixture was reacted at 70°C for 5 h under nitrogen protection, filtered, and the filtrate was extracted with EA. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford 100-40 as a brown oil (670 mg, 99%). m / z (ESI) [M+H] + =263.1.

[0254] Comparative Compound 1:

[0255] Compound 1-42 (2.0 g, 8.25 mmol) and compound 8-2 (2.27 g, 9.9 mmol) were placed in a round-bottom flask, HATU (4.7 g, 12.38 mmol) was added, and the mixture was dissolved in DMF (25 mL). Finally, DIPEA (4.3 mL, 24.75 mmol) was added and allowed to react at room temperature for 16 h. The mixture was quenched with water at 0°C and extracted with EA (100 mL). The EA layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA:petroleum ether = 1:10-1:2). The resulting solid was recrystallized from petroleum ether and ethyl acetate to afford comparative compound 1 (1.48 g, 40%) as a white solid.

[0256] 1 H NMR (400MHz, CDCl3) δ9.04 (s, 1H), 8.62 (s, 1H), 8.50 (d, J = 8.2Hz, 1H), 8.34 (d, J = 8 .0Hz,1H),7.93(d,J=8.2Hz,1H),7.88-7.84(m,2H),7.72(d,J=2.3Hz,1H),7.60(d, J=7.0Hz,1H),7.53-7.45(m,2H),7.38(t,J=7.8Hz,1H),7.22-7.18(m,2H),7.09(t ,J=7.8Hz,1H),6.95(d,J=8.2Hz,1H),6.82(t,J=5.0Hz,1H),4.38(d,J=5.5Hz,2H).

[0257] m / z(ESI)[M+H] + =454.

[0258] Preparation Example 16 Preparation of Comparative Compound 2 (AOH-1996)

[0259] Compound AOH-1996-43 (2.0 g, 9.29 mmol), compound 8-2 (2.6 g, 11.15 mmol) and HATU (5.3 g, 13.94 mmol) were dissolved in DMF (25 mL), and then DIPEA (4.9 mL, 27.87 mmol) was added dropwise. The mixture was reacted at room temperature under nitrogen for 16 h, and EA (100 mL) was added for extraction. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2), and recrystallized (EA / petroleum ether) to obtain an off-white solid AOH-1996 (630 mg, 16%).

[0260] 1 H NMR(400MHz, CDCl3)δ8.42(d,J=7.4Hz,2H),8.38–8.31(m,1H),7.93(d,J=8.3Hz,1H),7.89–7 .82(m,1H),7.61(d,J=7.0Hz,1H),7.56–7.44(m,2H),7.44–7.37(m,1H),7.21(t,J=8.5Hz,1H) ,7.13(dd,J=11.3,4.2Hz,1H),7.05(t,J=7.2Hz,1H),6.92(d,J=7.2Hz,1H),6.79(s,1H),6.67 (dd,J=8.9,1.8Hz,1H),6.61–6.53(m,2H),4.36(d,J=5.5Hz,2H),3.72(s,3H).m / z(ESI)[M+H] + =427.2.

[0261] Biological testing

[0262] 2.1 Antitumor activity test

[0263] 2.1.1 Cell culture

[0264] 2.1.2 Cell culture medium and seeding density

[0265] Table 2 Cell culture conditions

[0266] 2.1.3 Cell recovery

[0267] a) Remove the cryovial from liquid nitrogen and place it in a 37°C water bath to rapidly thaw. Wipe the surface of the cryovial with 75% alcohol and transfer it to a biosafety cabinet.

[0268] b) Transfer the frozen cells to a 15 mL sterile centrifuge tube, add 10 times the volume of culture medium as the freezing medium, and centrifuge at 1,200 rpm for 5 minutes.

[0269] c) Discard the culture medium in the centrifuge tube and add an appropriate volume of fresh culture medium (see Table 2), resuspend the cells, transfer to a culture flask, and culture in a 37°C cell culture incubator. Change the medium the next day.

[0270] 2.1.4 Cell passaging

[0271] Adherent cells:

[0272] a) For adherent cells in logarithmic growth phase, discard the culture medium, add 8 mL PBS and wash twice, then discard the PBS.

[0273] b) Add 5 mL of digestion solution containing 0.25% trypsin and incubate at 37°C for 3-5 minutes. Observe under a microscope that most cells have become rounded.

[0274] c) Add 10 mL of culture medium to terminate digestion. Gently pipette to digest the cells and prepare a cell suspension for passage and experiment.

[0275] Suspension cells:

[0276] a) Collect cells directly by centrifugation.

[0277] 2.1.5 Cell plating

[0278] a) Preparation of cell suspension

[0279] i. Remove the culture medium from the culture flask;

[0280] ii. Rinse the cells twice with PBS;

[0281] iii. Add trypsin to digest, stop digestion in the culture medium, and collect by centrifugation; suspended cells can be directly collected;

[0282] iv. Resuspend in culture medium, count and adjust to the appropriate concentration.

[0283] b) Add the cell suspension to a 384-well plate at a volume of 40 μL per well. See Table 2 for cell numbers.

[0284] c) Culture in a 37°C, 5% CO2 incubator for 48 hours.

[0285] 2.2 Compound treatment

[0286] 2.2.1 Compound dilution

[0287] a) Prepare gradient dilution solutions of positive compounds and test compounds: Prepare 10 mM stock solution of each compound in DMSO. Then perform 3-fold serial gradient dilutions in DMSO, with a total of 10 concentration points.

[0288] 2.2.2 Cell inoculation

[0289] a) Transfer 40 nL of compound using the Echo550 system; add the cell suspension to the cell culture plate at a volume of 40 μL per well.

[0290] b) Transfer the cells to an incubator and incubate for 48 hours.

[0291] 2.3 CTG detection

[0292] a) Take out the cell culture plate and equilibrate it at room temperature for half an hour.

[0293] b) Prepare the CTG assay working solution according to the Promega kit product instructions and equilibrate it in an incubator to 25°C.

[0294] c) Transfer 30 μL of CellTiter-Glo to each well of the cell culture plate, incubate at room temperature for 15 minutes, and then read the chemiluminescence using Ensight.

[0295] 2.4 Data Analysis

[0296] The inhibition rate (% Inhibition) was calculated using the following formula:

[0297] Lum HC :1‰ DMSO control group cell readings

[0298] Lum Sample : Cell readout after compound addition

[0299] Lum LC :Blank culture medium reading

[0300] IC calculation using XLfit (5.3.1.3) 50 , the fitting formula is selected from formula 201: y=(A+((BA) / (1+((x / C)^D))))

[0301] A:Bottom

[0302] B:Top

[0303] C:IC 50 (Rel IC50)

[0304] D:Hill Slope

[0305] Table 3 Compounds inhibiting HL-60 activity

[0306] Test results showed that by replacing the methoxyphenyl group of comparative compound 2 (AOH-1996) with a specific bicyclic ring, or further replacing the oxygen atom between the two benzene rings with a sulfur atom, the anti-tumor (HL-60) activity of the compounds was significantly enhanced. For example, compound 8 (MTB-1956) exhibited three times the HL-60 inhibitory activity of comparative compound 2 (AOH-1996), while compounds 44, 90, and 93 exhibited approximately nine times the HL-60 inhibitory activity of comparative compound 2 (AOH-1996). However, in comparative compound 1 (MTB-1962), replacing the methoxyphenyl group of comparative compound 2 (AOH-1996) with a benzothiazole bicyclic ring resulted in significantly lower activity, 11 times lower than that of comparative compound 2 (AOH-1996). The para-amino group is generally susceptible to P450 oxidation, and fluorine substitution can improve its biostability. The test results showed that the antitumor activity (HL-60) of fluoride compound 99 and its methyl deuterated compound 100 was similar to that of the corresponding compounds 44 and 93.

[0307] Table 4 Compounds inhibit the activity of NCI-H522

[0308] The test results showed that the anti-tumor (NCI-H522) activity of compounds such as compound 8 (MTB-1956) was significantly enhanced by replacing the methoxyphenyl group of AOH-1996 with a specific bicyclic ring. In contrast, the activity of compound 1, which replaced the methoxyphenyl group of AOH-1996 with a benzothiazole bicyclic ring, was lower.

[0309] Broad-spectrum anti-tumor activity test:

[0310] According to the above test method, different tumor cell lines were used to test the anti-tumor activity of the compound and AOH-1996 (see Table 5):

[0311] Test results showed that compound 44 (MTB-1970), compound 99, and their methyl-deuterated derivatives, compound 93 (MTB-1993), and compound 100 (MTB-2006), exhibited 5-10 times greater inhibitory activity against all tumor cells tested than AOH-1996. Furthermore, at the highest tested dose of 10 μM, they showed no significant cytotoxicity against normal human peripheral blood mononuclear cells (PBMCs), demonstrating significant development potential. Compound 100 (MTB-2006), in which the hydrogen at the para-amino position of compound 93 (MTB-1993) was replaced with fluorine, exhibited comparable antitumor activity to compound 93 (MTB-1993).

[0312] Pharmacokinetic studies

[0313] Pharmacokinetic study of compound 2 (AOH-1996), compound 100 (MTB-2006) and compound 99 after single oral administration in female CB-17 SCID (B-Ces1c KO) mice

[0314] 1. Research methods and experimental design

[0315] 1.1 Test system

[0316] 1.2 Experimental Design

[0317] Before the first administration, the animals were divided into groups according to their body weight. The animals were weighed before administration and the administration volume was calculated based on their body weight.

[0318] 2. Sample collection

[0319] 2.1. Drug delivery preparations

[0320] After each preparation was prepared, two portions of the clear dosing solution (upper and bottom layers) or three portions of the suspension (upper, middle, and bottom layers) were collected and analyzed by HPLC-UV to examine the accuracy of the preparation concentration.

[0321] 2.2 Sample Collection and Processing

[0322] Plasma samples were collected at 0.167, 0.333, 0.5, 1, 2, 4, 6, and 12 h after administration.

[0323] Whole Blood Collection and Plasma Sample Preparation: Whole blood was collected from the saphenous vein of each animal (approximately 0.02 mL per time point) in pre-chilled commercial blood collection tubes (containing EDTA-K2 as an anticoagulant). Within 10 seconds of blood collection, stabilizer was added at a ratio of whole blood to 400 mM DDV (v:v, 19:1) and the tubes were placed on wet ice until centrifugation. Each blood collection was kept on wet ice prior to centrifugation. Plasma samples were prepared by centrifugation at approximately 4°C, 3,200 g for 10 minutes. Blood samples were then transferred to low-binding tubes, flash-frozen on dry ice, and stored at -60°C or below until LC-MS / MS analysis. Sample processing was performed on wet ice.

[0324] 3. Sample analysis

[0325] 3.1. Drug delivery preparations

[0326] The concentration of the drug product should be determined by HPLC-UV. The calibration curve should include at least six concentration levels. Each sample should be analyzed in duplicate. The actual measured concentration should be within 80% to 120% of the theoretical prepared concentration. Otherwise, the actual dose should be used to calculate the pharmacokinetic parameters.

[0327] 3.2 Bioanalytical Method Development and Sample Analysis

[0328] Under Non-GLP conditions, an LC-MS / MS method was established for the quantitative determination of the test compounds in biological matrices.

[0329] Table 6. Pharmacokinetic data of AOH-1996, Compound 99, and MTB-2006

[0330] The results of the pharmacokinetic study (Table 6) show that compound 99 and compound 100 (MTB-2006) have better biostability. The deuteration of the methyl group further improves their biostability. The drug exposure of compound MTB-2006 is nearly three times that of AOH-1996. The exposure of the deuterated compound MTB-2006 is also significantly increased (32.5%) compared to compound 99. max The half-life and duration of MTB-2006 were significantly improved compared with those of compound AOH-1996, and MTB-2006 was even better.

[0331] The above test results show that the exemplary compounds have 5-10 times the inhibitory activity against various tumor cells in vitro compared to comparative compound 2 (AOH-1996), and their biostability and drug exposure are significantly improved, indicating greater development potential and clinical value.

[0332] The above is an illustrative description of the specific embodiments of the present invention. However, the present invention is not limited to the above specific embodiments. Any modifications, equivalent substitutions, etc. made within the spirit and principles of the present invention should be included in the scope of the present invention.

Claims

1. A compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, in: B is selected from substituted or unsubstituted naphthyl, substituted or unsubstituted quinolyl, or substituted or unsubstituted isoquinolyl; alternatively, B is selected from substituted or unsubstituted naphthyl; alternatively, B is naphthyl; alternatively, the substituents in the substituted naphthyl, substituted quinolyl, and substituted isoquinolyl are each independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl, halogen, or halogenated C 1-6 Alkyl; optionally, the substituents are each independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, phenyl, F, Cl, Br, I, or -CF3; R is each independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -NH2, -CN, halogen (such as F, Cl, Br, I), -OH, or -OR', wherein R' is C 1-6 Alkyl, C 3-7 Cycloalkyl, or substituted or unsubstituted C 6-10 aryl; optionally, the alkyl is methyl, ethyl, n-propyl, or isopropyl; the cycloalkyl is cyclopropyl; the substituents of the substituted aryl and the substituted heteroaryl are each independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl, halogen, or halogenated C 1-6 Alkyl; optionally, the substituents are each independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, phenyl, F, Cl, Br, I, or -CF3; n is an integer selected from 0-3; optionally, n is 0; Y is O or S; preferably, Y is O; preferably, Y is S; Ar is a carbocyclic fused phenyl, a heterocyclic fused phenyl, a carbocyclic fused heteroaryl, or a heterocyclic fused heteroaryl; optionally, Ar is a carbocyclic fused phenyl, or a heterocyclic fused phenyl; the carbocyclic ring is a saturated or non-aromatic unsaturated 5-membered or 6-membered carbocyclic ring, the heterocyclic ring is a saturated or non-aromatic unsaturated 5-membered or 6-membered heterocyclic ring containing 1 or 2 N atoms, or 1 or 2 O atoms, or 1 or 2 S atoms, and the heteroaryl is a 6-membered heteroaryl ring containing 1 or 2 N atoms; optionally, Ar is a heterocyclic fused phenyl, and the heterocyclic ring is a saturated or non-aromatic unsaturated 5-membered or 6-membered heterocyclic ring containing 1 or 2 O atoms; The carbocyclic fused phenyl, heterocyclic fused phenyl, carbocyclic fused heteroaryl, and heterocyclic fused heteroaryl are optionally substituted by one or more substituents, each of which is independently selected from C 1-6 Alkyl, C 3-7 Cycloalkyl, OH, CN, NH2, mono- or di-C 1-6 Alkylamino, halogen, or halogenated C 1-6 alkyl; optionally, the carbocyclic-fused phenyl, heterocyclic-fused phenyl, carbocyclic-fused heteroaryl, and heterocyclic-fused heteroaryl are optionally each independently substituted by 1 or 2 substituents, and the substituents are each independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CF3, or -CF3; optionally, the substituents are each independently selected from -CH3, -F, -Cl, or -CF3.

2. The compound represented by formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, wherein: Ar is selected from the following groups: Among them, X 1 , X 2 , X 3 , X 4 , X 5 , and X 6 Each independently selected from H, C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, or isopropyl), C 3-7 Cycloalkyl (e.g., cyclopropyl), OH, CN, NH2, halogen (e.g., Cl, Br, I, or F), or halogenated C 1-6 alkyl; optionally, X 1 , X 4 , and X 5 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CF3, or -CF3, preferably X 1 and / or X 4 and / or X 5 is hydrogen; optionally, X 1 , X 4 , and X 5 Each independently selected from H, -CH3, -F, -Cl, or -CF3, preferably X 1 and / or X 4 and / or X 5 is hydrogen; optionally, X 2 , X 3 , and X 6 Each is independently selected from H, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CF3, or -CF3, preferably X 2 and / or X 3 and / or X 6 is hydrogen; optionally, X 2 , X 3 , and X 6 Each independently is H, or -CH3, preferably X 2 , X 3 , and X 6 In the embodiment, both or both are hydrogen.

3. The compound represented by formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, wherein: Ar is selected from the following groups: Wherein, X is independently selected from C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, or isopropyl), C 3-7 Cycloalkyl (e.g., cyclopropyl), OH, CN, NH2, halogen (e.g., Cl, Br, I, or F), or halogenated C 1-6 Alkyl; optionally, each X is independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, F, Cl, Br, I, -CH2CH3, or -CF3; optionally, each X is independently selected from -CH3, -F, -Cl, or -CF3; m is an integer from 0 to 3; optionally, m is 0, 1 or 2.

4. The compound of formula (I) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, wherein: The compound of formula (I) is selected from the following compounds: Wherein, X is independently selected from C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, or isopropyl), C 3-7 Cycloalkyl (e.g., cyclopropyl), -OH, -CN, -NH2, halogen (e.g., Cl, Br, I, or F), or halogenated C 1-6 Alkyl; optionally, each X is independently selected from methyl, ethyl, propyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH2CH3, or -CF3; optionally, each X is independently selected from -CH3, -F, -Cl, or -CF3; m is an integer from 0 to 3; optionally, m is 0, 1 or 2.

5. The compound represented by formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, wherein: The compound of formula (I) or its deuterated compound is selected from the following compounds: Compound 44, Compound 93, Compound 99 or Compound 100 is particularly preferred.

6. The compound of formula (I) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, wherein: The pharmaceutically acceptable salt is a salt formed by a compound of formula (I) or a deuterated compound thereof and an acid; the acid includes an inorganic acid or an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or carbonic acid; optionally, the organic acid includes formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, phthalic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid.

7. A method for preparing a compound of formula (I) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, the method comprising the following reaction route: The definitions of Ar, Y, R, n, and B in the above structural formulae are the same as those in any one of claims 1 to 6.

8. The following intermediate compounds:

9. A pharmaceutical composition comprising one or more of the compound of formula (I) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, and optionally a pharmaceutically acceptable excipient.

10. The pharmaceutical composition of claim 9, further comprising an anticancer drug other than the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof; preferably, the anticancer drug comprises a chemotherapeutic drug, or / and a targeted drug, or / and a nuclear drug; preferably, the anticancer drug comprises a chemotherapeutic drug such as cyclophosphamide; an antimetabolite such as fluorouracil, gemcitabine or capecitabine; an antibiotic such as doxorubicin; a platinum anticancer drug such as carboplatin, cisplatin or oxaliplatin; a botanical drug such as irinotecan, paclitaxel or taxotere; a targeted drug Such as afatinib, trastuzumab, imatinib, panitumumab, ramucirumab or osimertinib; immunotherapies such as pembrolizumab, nivolumab or pembrolizumab; ADC drugs such as gemtuzumab, emtansine trastuzumab, ruconazole tuzumab, detrastuzumab or Enhertu; dual-antibody drugs such as PD-1 / CTLA-4 dual-antibody Kaitanib or PD-(L)1 / VEGF dual-antibody Ivoside, nuclear drugs such as lutetium [177Lu] texivir pituitide injection (Pluvicto) or Lutathera.

11. Use of the compound of formula (I) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, or the pharmaceutical composition according to claim 9 in the preparation of a PCNA inhibitor.

12. Use of the compound of formula (I) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, or the pharmaceutical composition according to claim 9 in the preparation of a drug for preventing or treating cancer; optionally, the cancer includes, but is not limited to, lung cancer, melanoma, colon cancer, rectal cancer, prostate cancer, ovarian cancer, and leukemia; preferably, the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, or the pharmaceutical composition can also be used in combination with anticancer drugs other than these; preferably, the anticancer drugs include chemotherapeutic drugs, or / and targeted drugs, or / and nuclear drugs; preferably, the anticancer drugs include chemotherapeutic drugs such as cyclophosphamide; antimetabolites such as fluorouracil Pyrimidine, gemcitabine or capecitabine; antibiotics such as doxorubicin; platinum anticancer drugs such as carboplatin, cisplatin or oxaliplatin; botanical drugs such as irinotecan, paclitaxel or taxotere; targeted drugs such as afatinib, trastuzumab, imatinib, panitumumab, ramucirumab or osimertinib; immunotherapies such as pembrolizumab, nivolumab or pembrolizumab; ADC drugs such as gemtuzumab, emtansine trastuzumab, ruconazole trastuzumab, detrastuzumab or Enhertu; dual-antibody drugs such as PD-1 / CTLA-4 dual-antibody Kaitanib or PD-(L)1 / VEGF dual-antibody Ivoside, nuclear drugs such as lutetium [177Lu] texivirpitide injection (Pluvicto) or Lutathera.

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