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

Abstract

The invention relates to a compound as shown in a formula (I) as a PCNA inhibitor, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, a preparation method thereof, an intermediate compound thereof, a pharmaceutical composition thereof and an application thereof. The PCNA inhibitor and the pharmaceutical composition containing the PCNA inhibitor can be used for treating lung cancer, melanin cancer, colon cancer, rectal cancer, prostate cancer, ovarian cancer, leukemia and other cancers with PCNA overexpression.

Description

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, the expression of PCNA usually increases significantly because they need more DNA replication to maintain their rapid proliferation state. In addition, PCNA is also involved in DNA repair and damage response of tumor cells. PCNA can be used as a cancer marker as well as 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 less 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 effect of cancer treatment. Summary of the invention

[0006] The inventors have found that changing the methoxy benzene ring of AOH-1996 to a bicyclic ring, or further changing the oxygen (O) between the two benzene rings to sulfur (S), has a significant effect on its anti-tumor activity. For example, as shown in the following structural formula, MTB-1956 obtained by introducing cyclopentyl ether at the 3,4-position of the phenyl group of AOH-1996 has an activity of inhibiting human acute promyelocytic leukemia cells (HL-60) that is 3 times that of AOH-1996, while the activity of MTB-1962 introduced with the NS five-membered heterocycle at the 3,4-position is reduced by 32 times. The inventors have obtained more effective PCNA inhibitors through a large number of screenings, which improves the therapeutic effect of cancer. In addition, deuterating the hydrogen atoms therein can improve its pharmacokinetic properties.

[0007]

[0008] Therefore, the present invention provides a new PCNA inhibitor, which 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 in the above cancers.

[0009] The present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0010]

[0011] in:

[0012] 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 -CF 3 ;

[0013] 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, -NH 2 , -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 -CF 3 ;

[0014] n is an integer selected from 0-3; optionally, n is 0;

[0015] Y is O or S; preferably, Y is O; preferably, Y is S;

[0016] 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;

[0017] 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, NH 2 , single or dual 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, -CH 2 CF 3 , or -CF 3 ; Optionally, the substituents are each independently selected from -CH 3 , -F, -Cl, or -CF 3 .

[0018] Optionally, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0019] Ar is selected from the following groups:

[0020]

[0021] 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), C3-7 Cycloalkyl (e.g. cyclopropyl), OH, CN, NH 2 , halogen (such as 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, -CH 2 CF 3 , or -CF 3 , 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, -CH 3 , -F, -Cl, or -CF 3 , 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, -CH 2 CF 3 , or -CF 3 , preferably X 2 and / or X 3 and / or X 6 is hydrogen; optionally, X 2 , X 3 , and X 6 are each independently H, or -CH 3 , preferably X 2 , X 3 , and X 6 In the embodiment, both or both are hydrogen.

[0022] Optionally, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0023] Ar is selected from the following groups:

[0024]

[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 (such as Cl, Br, I, or F) or halogenated C 1-6 alkyl; optionally, X is independently selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, F, Cl, Br, I, -CH 2 CH 3 , or -CF 3 ; Optionally, X is each independently selected from -CH 3 , -F, -Cl, or -CF 3 ;

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

[0027] Optionally, 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) is selected from the following compounds:

[0029]

[0030]

[0031]

[0032] 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, -NH 2 , halogen (such as Cl, Br, I, or F), or halogenated C 1-6 alkyl; optionally, X is independently selected from methyl, ethyl, propyl, n-propyl, isopropyl, cyclopropyl, -F, -Cl, -Br, -I, -CH 2 CH 3 , or -CF 3 ; Optionally, X is each independently selected from -CH 3 , -F, -Cl, or -CF 3 ;

[0033] m is an integer from 0 to 3; optionally, m is 0, 1 or 2.

[0034] Optionally, in the compound represented by the above formula (I), or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof,

[0035] The compound of formula (I) or its deuterated compound is selected from the following compounds:

[0036]

[0037]

[0038]

[0039]

[0040]

[0041] Compound 44, Compound 93, Compound 99 or Compound 100 is particularly preferred.

[0042] Optionally, in the compound represented by the above formula (I), or its pharmaceutically acceptable salt, or their deuterated compound, 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.

[0043] The pharmaceutically acceptable salts can be prepared by conventional methods in the art.

[0044] 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.

[0045] The compound represented by formula (I) of the present invention, its pharmaceutically acceptable salt, or its deuterated compound may exist in the form of amorphous, crystalline, solvent compound (such as hydrate), etc.

[0046] 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, the method comprising the following reaction route:

[0047]

[0048] The definitions of Ar, Y, R, n and B in the above structural formulae are the same as those described above.

[0049] The reaction process of this reaction route is as follows: Compound Ia as an aniline precursor and Compound Ib as a glycine derivative are subjected to condensation reaction in an organic solvent (such as DMF) under the action of a condensation agent (polypeptide condensation reagent (HATU)) and a catalyst (such as N,N-diisopropylethylamine (DIPEA)) to obtain the target product Compound I or its deuterated compound.

[0050] 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.

[0051]

[0052] The present invention also provides a pharmaceutical composition, which comprises one or more of the above-mentioned compound of formula (I) according to the present invention, or a pharmaceutically acceptable salt thereof, or a deuterated compound thereof, and optionally a pharmaceutically acceptable excipient. Optionally, the pharmaceutical composition further contains 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 drugs include, but are not limited to, chemotherapeutic 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, immunotherapy such as pamuzumab, nivolumab and pembrolizumab, ADC drugs such as gemtuzumab, trastuzumab enmetuzumab, ruconazole, 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.

[0053] The present invention also provides the use of the compound of formula (I) or its pharmaceutically acceptable salt or deuterated compound thereof or the pharmaceutical composition in the preparation of PCNA inhibitors.

[0054] The present invention also provides the use of the above-mentioned compound of formula (I) of the present invention, or its pharmaceutically acceptable salt, or their deuterated compound, 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, cancer with overexpression of PCNA.

[0055] The present invention also provides a method for preventing or treating cancer, comprising administering to an individual in need thereof an 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.

[0056] The compound of formula (I) or its pharmaceutically acceptable salt or its deuterated compound described herein, or the pharmaceutical composition containing the compound of formula (I) or its pharmaceutically acceptable salt or its deuterated compound can be used alone or in combination with other drugs. The appropriate combination can produce a synergistic effect, improve the therapeutic effect of the above-mentioned cancer, or reduce the toxic and side effects of the drug, or can also reduce the toxic and side effects while producing a synergistic effect. The 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, chemotherapeutic 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, immunotherapy such as pamuzumab, nivolumab and pembrolizumab, ADC drugs such as gemtuzumab, trastuzumab enmetuzumab, ruconazole, 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

[0057] The specific embodiments of the present invention are described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate the present invention by way of example and are not used to limit the present invention.

[0058] Preparation of compounds

[0059] Preparation Example 1 Preparation of Compound 8 (MTB-1956)

[0060] Compound 8-2:

[0061]

[0062] Compound 8-1 (40 g, 232.32 mmol) in SOCl 2 (150 mL) was refluxed for 2 h, and SOCl was distilled off 2, add 1,4-dioxane (20mL) and evaporate twice, the residue is dissolved in 1,4-dioxane (100mL). Dissolve NaOH (18.6g, 464.64mmol) in water (232mL), then add glycine (17.4g, 232.32mmol) in sodium hydroxide aqueous solution, after dissolving, add dropwise to the above reaction solution at 0°C, the resulting reaction solution is heated to room temperature and reacted for 16h, EA (200mL) is added for extraction, the aqueous phase is adjusted to pH 2 with 5N HCl, the solid is precipitated, filtered, washed with water, and dried to obtain compound 8-2 (41.8g, 79%) as a light yellow solid.

[0063] 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).

[0064] m / z(ESI)[M+H] + =230.1.

[0065] Compound 8-4:

[0066]

[0067] Compound 8-3 (5 g, 30.49 mmol) was dissolved in THF (60 mL) and NaHCO 3 (5.1 g, 60.98 mmol), H was added dropwise at 0 °C 2 O 2 (30%, 6.2 mL, 60.98 mmol), react at room temperature for 16 h, add saturated sodium bisulfite solution to quench, extract with EA (200 mL), dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and chromatograph on a silica gel column (methanol:CH 2 Cl 2 =1:30-1:20) to give compound 8-4 (4.1 g, 99%) as a white solid.

[0068] 1 H NMR (400 MHz, CDCl 3)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).

[0069] m / z(ESI)[MH] - =135.1.

[0070] Compound 8-6:

[0071]

[0072] Compound 8-5 (3.5 g, 25.09 mmol), compound 8-4 (4.1 g, 30.11 mmol), K 2 CO 3 (3.8 g, 27.60 mmol) was added to DMF (50 mL), protected by nitrogen, reacted at 100°C for 16 h, cooled, 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.

[0073] m / z(ESI)[M+Na] + =279.9.

[0074] Compound 8-7:

[0075]

[0076] Compound 8-6 (6.4 g, 24.9 mmol) was dissolved in a mixed solvent of EA (50 mL) and MeOH (50 mL), 0.6 g of 10% Pd / C and hydrogen were added, and the mixture was reacted at room temperature for 16 h. The mixture was filtered through celite, washed with EA, and the filtrate was concentrated to obtain compound 8-7 (5.6 g, 99%) as a gray oil.

[0077] 1 H NMR (400 MHz, CDCl 3 )δ6.98-6.89(m,1H),6.88-6.83(m,1H),6.81(dd,J=7.8,1.6Hz,1H),6.79-6.74(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).

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

[0079] Compound 8:

[0080]

[0081] 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, 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 = 2:10-1:2), and then recrystallized (EA / petroleum ether) to obtain compound 8 (2.2 g, 57%) as an off-white solid.

[0082] 1 H NMR (400 MHz, CDCl 3 )δ8.53(s,1H),8.39(dd,J=7.9,1.1Hz,1H),8.34(d,J=8.4Hz,1H),7.92(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).

[0083] m / z(ESI)[M+H] + =439.1.

[0084] Preparation Example 2 Preparation of Compound 9 (MTB-1959)

[0085] Compound 9-9:

[0086]

[0087] Compound 9-8 (5 g, 25.12 mmol), biboric acid pinacol ester (7.7 g, 30.14 mmol), potassium acetate (7.4 g, 75.36 mmol), Pd(dppf)Cl 2(919 mg, 1.26 mmol), Dioxane (50 mL) was added, and the reaction was carried out 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.

[0088] 1 H NMR (400 MHz, CDCl 3 )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).

[0089] Compounds 9-10:

[0090]

[0091] Compound 9-9 (6 g, 24.38 mmol) was dissolved in THF (50 mL) and NaHCO 3 (4.1 g, 48.76 mmol), and then H was added dropwise at 0 °C. 2 O 2 (30%, 5 mL, 48.76 mmol), react at room temperature for 3 h, add saturated sodium bisulfite solution to quench, add EA (100 mL) to extract, dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography (methanol:CH 2 Cl 2 =1:30-1:20) to give compound 9-10 (3.23 g, 97%) as a colorless oil.

[0092] 1 H NMR (400 MHz, CDCl 3 )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).

[0093] m / z(ESI)[M+H] + =137.1.

[0094] Compound 9-11:

[0095]

[0096] Compound 8-5 (3.0 g, 21.57 mmol), compound 9-10 (3.23 g, 23.72 mmol) and K 2 CO 3 (3.23 g, 23.72 mmol) was added to DMF (40 mL), and the reaction was carried out at 110° C. for 16 h under nitrogen protection. The mixture was 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 give compound 9-11 (5.08 g, 92%) as a yellow solid.

[0097] Compounds 9-12:

[0098]

[0099] 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.

[0100] 1 H NMR (400 MHz, CDCl 3 )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=7.9,1.5Hz,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).

[0101] m / z(ESI)[M+H] + =228.1.

[0102] Compound 9:

[0103]

[0104] 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, purified by silica gel column chromatography (EA: petroleum ether = 1:10-1:2), and then recrystallized (EA / petroleum ether) to obtain compound 9 (1.8 g, 47%) as a yellow solid.

[0105] 1 H NMR (400 MHz, CDCl 3 )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).

[0106] m / z(ESI)[M+H] + =439.1.

[0107] Preparation Example 3 Preparation of Compound 15 (MTB-1958)

[0108] Compound 15-14:

[0109]

[0110] Compound 8-5 (5.0 g, 35.44 mmol), compound 15-13 (5.4 g, 38.98 mmol) and K 2 CO 3 (5.4 g, 38.98 mmol) was added to DMF (50 mL), and the reaction was carried out at 100° C. for 16 h under nitrogen protection. The mixture was cooled to room temperature, and EA (200 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 = 1:10-1:3) to give compound 15-14 (9.0 g, 98%) as a yellow solid.

[0111] Compound 15-15:

[0112]

[0113] 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.

[0114] 1 H NMR (400 MHz, CDCl 3 )δ6.98-6.90(m,1H),6.81(dt,J=7.8,1.5Hz,2H),6.76-6.66(m,2H),6.5 6(d,J=2.4Hz,1H), 6.44(dd,J=8.4,2.5Hz,1H), 5.95(s,2H), 3.82(s,2H).

[0115] m / z(ESI)[M+H] + =230.1.

[0116] Compound 15:

[0117]

[0118] 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, 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 compound 15 (0.7 g, 18%) as an off-white solid.

[0119] 1 H NMR (400 MHz, CDCl 3)δ8.50(s,1H),8.39(d,J=6.9Hz,1H),8.36-8.29(m,1H),7.92(d,J=8.2Hz,1H),7.88-7.81(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=7.8,1.3H z,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).

[0120] m / z(ESI)[M+H] + =441.2.

[0121] Preparation Example 4 Preparation of Compound 17 (MTB-1961):

[0122] Compound 17-17:

[0123]

[0124] Compound 8-5 (3.0 g, 21.26 mmol), compound 17-16 (3.6 g, 23.39 mmol) and K 2 CO 3 (3.23 g, 23.39 mmol) was added to DMF (50 mL), and the reaction was carried out at 100°C for 16 h under nitrogen protection. The mixture was cooled to room temperature, 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 = 1:10-1:3) to give compound 17-17 (5.8 g, 99%) as a yellow solid.

[0125] m / z(ESI)[M+H] + =274.0.

[0126] Compounds 17-18:

[0127]

[0128] 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%).

[0129] m / z(ESI)[M+H] + =244.1.

[0130] Compound 17:

[0131]

[0132] 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, 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 compound 17 (0.5 g, 13%) as a yellow solid.

[0133] 1 H NMR (400 MHz, CDCl 3 )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(t d,J=7.8,1.2Hz,1H),7.04-6.98(m,1H),6.90(s,1H),6.83(dd,J=11.1,4.9Hz,2H),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).

[0134] m / z(ESI)[M+H] + =455.1.

[0135] Preparation Example 5 Preparation of Compound 20 (MTB-1965):

[0136] Compounds 20-21:

[0137]

[0138] 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, and potassium acetate (6.98 g, 71.1 mmol) and PdCl 2 (dppf) (0.87 g, 1.19 mmol) was dissolved in 1,4-dioxane (60 mL). The reaction was stirred at 90°C for 3 h. Water was added at 0°C to quench the mixture, and the mixture was extracted with EA (100 mL). The mixture was 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 obtain compound 20-21 (2.36 g, 38%) as a yellow solid.

[0139] 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).

[0140] m / z(ESI)[M+H] + =259.

[0141] Compounds 20-22:

[0142]

[0143] Compound 20-21 (2.36 g, 9.14 mmol) was placed in a round-bottom flask, 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, and the mixture 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:CH 2 Cl 2 =1:30-1:20) to give compound 20-22 (1.0 g) as a white solid, which was used directly in the next step.

[0144] 1 H NMR (400MHz, DMSO) δ9.27 (s, 1H), 7.94 (s, 1H), 7.40 (d, J = 8.6 Hz, 1H), 6.82 ( d, J = 2.2 Hz, 1H), 6.69 ( dd, J = 2.2, 8.6 Hz, 1H), 3.72 ( s, 3H).

[0145] m / z(ESI)[M+H] + =149.

[0146] Compounds 20-23:

[0147]

[0148] 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, extracted with EA (100 mL), and 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.

[0149] 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).

[0150] m / z(ESI)[M+H] + =270.

[0151] Compounds 20-24:

[0152]

[0153] Compound 20-23 (1.7 g, 6.31 mmol) was placed in a round-bottom flask, EA (15 mL) and MeOH (10 mL) were added, and then 10% Pd / C (0.2 g) was added, and hydrogen was introduced. The reaction was carried out at room temperature for 16 h, filtered through diatomaceous earth, concentrated, and purified by silica gel column chromatography (methanol:CH 2 Cl 2 =1:30-1:10) to give compound 20-24 (1.5 g, 99%) as a foamy solid.

[0154] 1H 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).

[0155] m / z(ESI)[M+H] + =240.

[0156] Compound 20:

[0157]

[0158] 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, dissolved with DMF (20 mL), and finally DIPEA (3.4 mL, 19.44 mmol) was added, and the mixture was reacted at room temperature for 16 h. Water was added to quench at 0°C, and the mixture was 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, which was recrystallized with petroleum ether and ethyl acetate to obtain compound 20 (0.9 g, 31%) as a white solid.

[0159] 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=2.2,8.6Hz,1H), 6.81(dd,J=1.4,7.8Hz,1H), 4.19(d,J=6.0Hz,2H), 3.77(s,3H).

[0160] m / z(ESI)[M+H] + =451.

[0161] Preparation Example 6 Preparation of Compound 21 (MTB-1963):

[0162] Compounds 21-26:

[0163]

[0164] 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 mixture was reacted at 100°C overnight, cooled to room temperature, quenched with water at 0°C, extracted with EA (100 mL), and 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.

[0165] 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).

[0166] m / z(ESI)[M+H] + =270.

[0167] Compounds 21-27:

[0168]

[0169] Compound 21-26 (5.2 g, 19.31 mmol) was placed in a round-bottom flask, EA (45 mL) and MeOH (30 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 diatomaceous earth, concentrated, and dried in vacuo to obtain a foamy solid compound 21-27 (4.6 g, 99%).

[0170] 1H 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).

[0171] m / z(ESI)[M+H] + =240.

[0172] Compound 21:

[0173]

[0174] 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, dissolved with DMF (30 mL), and finally DIPEA (5.5 mL, 31.59 mmol) was added, and the mixture was reacted at room temperature for 16 h. Water was added to quench at 0°C, and the mixture was 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, which was recrystallized from ethyl acetate to obtain compound 21 (180 mg, 21%) as a white solid.

[0175] 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).

[0176] m / z(ESI)[M+H] + =451.

[0177] Preparation Example 7 Preparation of Compound 23 (MTB-1957)

[0178] Compound 23-29:

[0179]

[0180] Compound 23-28 (5 g, 23.8 mmol, pinacol diboron (9.1 g, 35.7 mmol), potassium acetate (7.0 g, 71.4 mmol) and Pd(dppf)Cl 2 (871 mg, 1.19 mmol) was added with Dioxane (60 mL), and the reaction was carried out 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.

[0181] 1 H NMR (400 MHz, CDCl 3 )δ8.16(s,1H),7.67(d,J=8.3Hz,1H),7.32(d,J=8.3Hz,1H),7.04(d,J=3.1Hz,1H),6.50(d,J=3.1Hz,1H),3.79(s,3H),1.37(s,12H).

[0182] Compounds 23-30:

[0183]

[0184] Compound 23-29 (5.9 g, 22.94 mmol) was dissolved in THF (50 mL) and NaHCO 3 (3.8 g, 45.88 mmol), and then H was added dropwise at 0°C. 2 O 2 (30%, 4.7 mL, 45.88 mmol), react at room temperature for 3 h, add saturated sodium bisulfite solution to quench, add EA (100 mL) to extract, the organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (methanol: CH 2 Cl 2 =1:30-1:20) to give compound 23-30 (1.38 g, 41%) as a yellow solid.

[0185] 1 H NMR (400 MHz, CDCl 3)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).

[0186] m / z(ESI)[M+H] + =148.1.

[0187] Compound 23-31:

[0188]

[0189] Compound 8-5 (1.2 g, 8.52 mmol), compound 23-30 (1.38 g, 9.37 mmol) and K 2 CO 3 (1.3 g, 9.37 mmol) was added to DMF (30 mL), and the reaction was carried out at 110° C. for 16 h under nitrogen protection. The mixture was cooled to room temperature, and 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 = 1:10-1:3) to give compound 23-31 (2.08 g, 91%) as a yellow solid.

[0190] m / z(ESI)[M+H] + =269.1.

[0191] Compound 23-32:

[0192]

[0193] 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.

[0194] m / z(ESI)[M+H] + =239.1.

[0195] Compound 23:

[0196]

[0197] 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, and EA (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After purification by silica gel column chromatography (EA: petroleum ether = 1:10-1:2), the white solid compound 23 (1.7 g, 50%) was obtained by recrystallization (EA / petroleum ether).

[0198] 1 H NMR (400 MHz, CDCl 3 )δ8.54(s,1H),8.42(d,J=7.1Hz,1H),8.35(d,J=8.3Hz,1H),7.90(d,J=8.3Hz,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),7.30-7.2 4(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,1 H), 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).

[0199] m / z(ESI)[M+H] + =450.1.

[0200] Preparation Example 8 Preparation of Compound 24 (MTB-1960):

[0201] Compound 24-34:

[0202]

[0203] Compound 24-33 (3.4 g, 13.22 mmol) was dissolved in THF (50 mL) and NaHCO 3 (2.2 g, 26.44 mmol), then H was added dropwise at 0°C. 2 O 2 (30%, 2.7 mL, 26.44 mmol), react at room temperature for 3 h, add saturated sodium bisulfite solution to quench, add EA (100 mL) to extract, the organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (methanol: CH2 Cl 2 =1:30-1:20) to give yellow oily compound 24-34 (1.9 g, 97%).

[0204] 1 H NMR (400 MHz, CDCl 3 )δ7.45(d,J=8.4Hz,1H),6.93(d,J=3.1Hz,1H),6.77(d,J=2.2Hz,1H),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).

[0205] m / z(ESI)[M+H] + =148.1.

[0206] Compounds 24-35:

[0207]

[0208] Compound 8-5 (1.5 g, 10.76 mmol), compound 24-34 (1.9 g, 12.91 mmol) and K 2 CO 3 (1.6 g, 11.8 mmol) was added to DMF (30 mL), and the reaction was carried out at 110° C. for 16 h under nitrogen protection. The mixture was cooled to room temperature, and EA (100 mL) was added for extraction. 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 a yellow solid 24-35 (2.4 g, 83%).

[0209] m / z(ESI)[M+H] + =269.1.

[0210] Compound 24-36:

[0211]

[0212] 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 mixture was reacted 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%).

[0213] m / z(ESI)[M+H] + =239.1.

[0214] Compound 24:

[0215]

[0216] 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, and EA (100 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. It 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.

[0217] 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).

[0218] m / z(ESI)[M+H] + =450.1.

[0219] Preparation Example 9 Preparation of Compound 44

[0220]

[0221]

[0222] 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%).

[0223] 1 H NMR (400 MHz, CDCl 3 )δ8.68(s,1H),8.38(d,J=7.5Hz,1H),8.33(d,J=8.4Hz,1H),7.90(d,J=8.1Hz,1 H),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,1H),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.

[0224] Preparation Example 10 Preparation of Compound 90:

[0225]

[0226] Compound 90 was prepared using the method for preparing compound 44.

[0227] m / z(ESI)[M+H] + =469.1.

[0228] Preparation Example 11 Preparation of Compound 93 (MTB-1993):

[0229]

[0230] Using the method for preparing compound 44, compound 93 was prepared from compound 93-38.

[0231] 1 H NMR (400 MHz, CDCl 3)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.

[0232] m / z(ESI)[M+H] + =457.2.

[0233] Preparation Example 12 Preparation of Compound 99

[0234]

[0235] 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%). 1 H NMR (400 MHz, CDCl 3)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.

[0236] Preparation Example 13 Preparation of Compound 100 (MTB-2006)

[0237]

[0238] 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%).

[0239] 1 H NMR (400 MHz, CDCl 3 )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,1H),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.

[0240] Preparation Example 14 Preparation of other analogs

[0241]

[0242] Using a similar method, the target products shown in Table 1 were prepared by condensation of compounds Ia and Ib.

[0243] Table 1

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250] Preparation Example 15 Preparation of Comparative Compound 1 (MTB-1962)

[0251] Compound 1-39:

[0252]

[0253] 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, and potassium acetate (20.63 g, 210.2 mmol) and PdCl were added. 2 (dppf) (2.56 g, 3.5 mmol) 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.

[0254] m / z(ESI)[M+H] + =262.

[0255] Compound 1-40:

[0256]

[0257]

[0258] Compound 1-39 (6.0 g, 22.98 mmol) was placed in a round-bottom flask, 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, and the mixture was stirred at room temperature for 3 h. The mixture was quenched with saturated sodium bisulfite, extracted with EA (100 mL), the EA layer was washed once with water, once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and recrystallized with petroleum ether and ethyl acetate to obtain 2.66 g of yellow-white solid compound 1-40, which was used directly in the next step.

[0259] 1 H 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).

[0260] m / z(ESI)[M+H] + =152.

[0261] Compound 1-41:

[0262]

[0263] 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, extracted with EA (100 mL), and 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.

[0264] 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).

[0265] m / z(ESI)[M+H] + =273.

[0266] Compound 1-42:

[0267]

[0268] 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, and hydrogen was introduced. The mixture was reacted at room temperature for 16 h, filtered through diatomaceous earth, concentrated, and dried in vacuo to obtain yellow oily compound 1-42 (4.28 g), which was directly used in the next step.

[0269] 1 H 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).

[0270] m / z(ESI)[M+H] + =243.

[0271] Preparation of Compound 44-37

[0272] Compound 44-37-2

[0273]

[0274] Compound 44-37-1 (10 g, 53.46 mmol), 1,2-dibromoethane (30.1 g, 160.38 mmol), potassium carbonate (11.1 g, 80.19 mmol) were added to acetonitrile (150 mL), and the mixture was reacted at 85 °C overnight under nitrogen protection. 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 colorless oil 44-37-2 (7.4 g, 47%).

[0275] 1 H NMR (400 MHz, CDCl 3 )δ7.40(d,J=7.9Hz,1H),6.72(d,J=1.5Hz,1H),6.69(dd,J=7.9,1.5Hz,1H),4.32(t,J=6.6Hz,2H),3.67(t,J=6.6Hz,2H),2.31(s,3H).

[0276] 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.

[0277] Compound 44-37-3

[0278]

[0279] 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.

[0280] Compound 44-37-4

[0281]

[0282] Compound 44-37-3 (3.06 g, 22.81 mmol) was dissolved in acetonitrile (45 mL), NBS (4.46 g, 25.08 mmol) was added in batches at 0°C, and then reacted at room temperature for 16 h. The solvent was distilled off, EA was added, and the organic phase was washed with saturated aqueous sodium bicarbonate solution, saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a light yellow solid 44-37-4 (4.8 g, 99%).

[0283] 1 H NMR (400 MHz, CDCl 3 )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).

[0284] Compound 44-37-5

[0285]

[0286] Compound 44-37-4 (4.8 g, 22.53 mmol), biboric acid pinacol ester (6.9 g, 27.03 mmol), potassium acetate (6.6 g, 67.59 mmol), Pd(dppf)Cl 2(824 mg, 1.13 mmol) was added with 1,4-dioxane (50 mL), and the reaction was carried out at 90 ° C for 6 h under nitrogen protection. The mixture was cooled to room temperature, EA and water were added, and the mixture was filtered through diatomaceous earth. The filtrate was extracted, 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 light yellow solid 44-37-5 (2.7 g, 46%).

[0287] Compound 44-37-6

[0288]

[0289] Compound 44-37-5 (2.7 g, 10.38 mmol) was dissolved in THF (20 mL) and NaHCO 3 (1.7 g, 20.76 mmol), and then 30% H 2 O 2 (2.1 mL, 20.76 mmol), react at room temperature for 3 h, add saturated sodium bisulfite solution to quench, add EA to extract, dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography (methanol:CH 2 Cl 2 =1:30-1:20) to give 44-37-6 (600 mg, 38%) as a colorless oil.

[0290] 1 H NMR (400 MHz, CDCl 3 )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).

[0291] Compound 44-37-7

[0292]

[0293] Compound 44-37-6 (600 mg, 4.0 mmol), 8-5 (470 mg, 3.33 mmol), and K 2 CO 3 (506 mg, 3.66 mmol) was added to DMF (10 mL), and the mixture was 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 44-37-7 (860 mg, 79%) as a brown oil.

[0294] Compound 44-37

[0295]

[0296] 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, filtered through celite, washed with EA, and the filtrate was concentrated to obtain brown oil 44-37 (764 mg, 99%). m / z (ESI) [M+H] + =242.1.

[0297] Preparation of Compound 93-38

[0298] Compound 93-38-2

[0299]

[0300] Compound 93-38-1 (10 g, 50.24 mmol) was dissolved in THF (100 mL), and n-BuLi (22 mL, 55.26 mmol, 2.5 M) was added dropwise at -78 °C under nitrogen protection, and stirred at -78 °C for 30 min. 3 I (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, quenched by adding saturated ammonium chloride solution, extracted with EA, and 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.

[0301] 1 H NMR (400 MHz, CDCl 3 )δ7.07(d,J=7.4Hz,1H),6.66(dd,J=7.4,1.4Hz,1H),6.63(d,J=1.4Hz,1H),4.55(t,J=8.7Hz,2H),3.16(t,J=8.7Hz,2H).

[0302] Compound 93-38-3

[0303]

[0304] Compound 93-38-2 (6.3 g, 45.92 mmol) was weighed in a round-bottom flask, acetonitrile (90 mL) was added, and NBS (9.0 g, 50.51 mmol) was added in batches, and the mixture was reacted at room temperature for 16 h. The solvent was removed by distillation, and the mixture was extracted with EA and water. The organic layer was washed with saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid 93-38-3 (9.9 g, 99%).

[0305] 1 H NMR (400 MHz, CDCl 3 )δ7.31(s,1H),6.67(s,1H),4.56(t,J=8.7Hz,2H),3.17(t,J=8.7Hz,2H).

[0306] Compound 93-38-4

[0307]

[0308] Compound 93-38-3 (9.9 g, 45.81 mmol), pinacol diboron (17.4 g, 68.72 mmol), potassium acetate (13.5 g, 137.43 mmol), Pd(dppf)Cl 2 (1.7 g, 29 mmol) was added with 1,4-dioxane (100 mL), and the reaction was carried out at 100 ° C for 16 h under nitrogen protection. 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%).

[0309] Compound 93-38-5

[0310]

[0311] Compound 93-38-4 (7.7 g, 29.26 mmol) was dissolved in THF (70 mL) and NaHCO 3 (4.9 g, 58.52 mmol), and then H was added dropwise at 0°C. 2 O 2 (6 mL, 58.52 mmol, 30%), react at room temperature for 16 h, add saturated sodium bisulfite solution to quench, add EA to extract, dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography (methanol:CH 2 Cl 2 =1:30-1:20) to obtain a light yellow solid 93-38-5 (2.4 g, 54%).

[0312] 1 H NMR (400 MHz, CDCl 3 )δ6.66(s,1H),6.56(s,1H),4.51(t,J=8.6Hz,2H),4.34(s,1H),3.13(t,J=8.6Hz,2H).

[0313] Compound 93-38-6

[0314]

[0315] Compound 93-38-5 (2.4 g, 15.67 mmol), 8-5 (2.0 g, 14.24 mmol), K 2 CO 3 (2.2 g, 15.67 mmol) was added to DMF (30 mL), and the reaction was carried out 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 a yellow solid 93-38-6 (1.9 g, 44%).

[0316] 1 H NMR (400 MHz, CDCl 3 )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, 1.0 Hz, 1H), 6.68 (s, 1H), 4.59 (t, J = 8.6 Hz, 2H), 3.18 (t, J = 8.6 Hz, 2H).

[0317] Compound 93-38

[0318]

[0319] To compound 93-38-6 (1.7 g, 6.20 mmol), NH 4 EtOH (40 mL) was added to Cl (2.0 g, 37.2 mmol), and then zinc powder (2.4 g, 37.2 mmol) was added in batches. The mixture was reacted at 80 °C for 16 h under nitrogen protection, filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (methanol: CH 2 Cl 2

[0320] =1:30-1:10) to obtain 93-38 (1.51 g, 99%) as a red oil. m / z (ESI) [M+H] + =245.1.

[0321] Preparation of Compound 99-39

[0322] Compound 99-39-2

[0323]

[0324] Compound 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 in batches at 0°C, and then reacted at room temperature for 2 h, quenched with water, extracted with EA, and 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 obtain a yellow solid 99-39-2 (320 mg, 33%). m / z (ESI) [M+H] + =290.1.

[0325] Compound 99-39

[0326]

[0327] Compound 99-39-2 (500 mg, 1.73 mmol) and NH 4 Cl (555 mg, 10.38 mmol) was added to THF (20 mL) and water (10 mL), and then zinc powder (673 mg, 10.38 mmol) was added in batches. Under nitrogen protection, the mixture was reacted at 70°C for 5 h, filtered, and the filtrate was extracted with EA. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain brown oily 99-39 (325 mg, 72%). m / z (ESI)

[0328] [M+H] + =260.1.

[0329] Preparation of Compound 100-40

[0330] Compound 100-40-1

[0331]

[0332] 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, quenched with water, extracted with EA, and 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 obtain a yellow solid 100-40-1 (750 mg, 39%) (determined according to the literature for the ortho-selective substitution reaction of the nitro group).

[0333] 1 H NMR (400 MHz, CDCl 3 )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).

[0334] 13 C NMR (101 MHz, CDCl 3 )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).

[0335] Compound 100-40

[0336]

[0337] Compound 100-40-1 (750 mg, 2.57 mmol), NH 4 Cl (825 mg, 15.42 mmol) was added to THF (20 mL), water (10 mL), and then zinc powder (1.0 g, 15.42 mmol) was added in batches. Under nitrogen protection, the mixture was reacted at 70°C for 5 h, filtered, and the filtrate was extracted with EA. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain brown oil 100-40 (670 mg, 99%). m / z (ESI) [M+H] + =263.1.

[0338] Comparative Compound 1:

[0339]

[0340] 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, dissolved with DMF (25 mL), and finally DIPEA (4.3 mL, 24.75 mmol) was added, and the mixture was reacted at room temperature for 16 h. Water was added to quench at 0°C, and the mixture was 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, which was recrystallized from petroleum ether and ethyl acetate to obtain a white solid comparative compound 1 (1.48 g, 40%).

[0341] 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).

[0342] m / z(ESI)[M+H] + =454.

[0343] Preparation Example 16 Preparation of Comparative Compound 2 (AOH-1996)

[0344]

[0345] 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 for 16 h under nitrogen protection. EA (100 mL) was added for extraction, and 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 an off-white solid AOH-1996 (630 mg, 16%).

[0346] 1 H NMR (400 MHz, CDCl 3 )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.

[0347] Biological Testing

[0348] 2.1 Antitumor activity test

[0349] 2.1.1 Cell culture

[0350] 2.1.2 Cell culture medium and seeding density

[0351] Table 2 Cell culture conditions

[0352] Cell lines Culture medium Seeding density (cells / well) HL-60 IMDM + 20% FBS 3200 NCI-H522 1640+10%FBS 1600

[0353] 2.1.3 Cell recovery

[0354] a) Take out the cell cryopreservation tube from liquid nitrogen and place it in a 37℃ water bath to thaw quickly. Wipe the surface of the cryopreservation tube with 75% alcohol and move it into a biosafety cabinet.

[0355] b) Transfer the frozen cells to a 15 mL sterile centrifuge tube, add 10 times the volume of the freezing medium into the culture medium, and centrifuge at 1,200 rpm for 5 min.

[0356] c) Discard the culture medium in the centrifuge tube, 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, and change the medium the next day.

[0357] 2.1.4 Cell passaging

[0358] Adherent cells:

[0359] a) For adherent cells in logarithmic growth phase, discard the culture medium, add 8 mL PBS and wash twice, then discard the PBS.

[0360] b) Add 5 mL of digestion solution containing 0.25% trypsin and place at 37°C for 3-5 minutes. Under a microscope, most cells become round in shape.

[0361] c) Add 10 mL of culture medium to terminate digestion, and gently pipette to digest the cells to make a cell suspension for passaging and experiments.

[0362] Suspension cells:

[0363] a) Collect the cells directly by centrifugation.

[0364] 2.1.5 Cell plating

[0365] a) Preparation of cell suspension

[0366] i. Remove the culture medium in the culture flask;

[0367] ii. Rinse the cells twice with PBS;

[0368] iii. Add trypsin to digest, and after the culture medium stops digestion, collect by centrifugation; collect suspended cells directly;

[0369] iv. Resuspend in culture medium, count and adjust to appropriate concentration.

[0370] b) Add the cell suspension to a 384-well plate at a volume of 40 μL per well. The cell numbers are shown in Table 2.

[0371] c) 37°C, 5% CO 2 Incubate in an incubator for 48 hours.

[0372] 2.2 Compound treatment

[0373] 2.2.1 Compound dilution

[0374] a) Prepare gradient dilution solutions of positive compounds and test compounds: Prepare 10 mM stock solution of each compound with DMSO. Then perform 3-fold continuous gradient dilution with DMSO, with a total of 10 concentration points for the compound.

[0375] 2.2.2 Cell inoculation

[0376] a) Transfer 40 nL of compound using the Echo550 system; add cell suspension to the cell culture plate at a volume of 40 μL per well.

[0377] b) Transfer the cells to an incubator and incubate for 48 hours.

[0378] 2.3CTG detection

[0379] a) Take out the cell culture plate and equilibrate it at room temperature for half an hour.

[0380] b) Prepare the CTG detection working solution according to the Promega kit product instructions and place it in a constant temperature box to equilibrate to 25 degrees Celsius.

[0381] c) Transfer 30 μL of CellTiter-Glo to each well of the cell culture plate, incubate at room temperature for 15 minutes, and read the chemiluminescence using Ensight.

[0382] 2.4 Data Analysis

[0383] The inhibition rate (%Inhibition) was calculated using the following formula:

[0384]

[0385] Lum HC :1‰ DMSO control group cell reading

[0386] Lum Sample : Cell readout after compound addition

[0387] Lum LC :Blank medium reading

[0388] IC calculation using XLfit (5.3.1.3) 50 , the fitting formula is selected from formula 201: y = (A + ((BA) / (1 + ((x / C) ^ D))))

[0389] A:Bottom

[0390] B:Top

[0391] C:IC 50 (Rel IC50)

[0392] D:Hill Slope

[0393] Table 3 Compounds inhibiting HL-60 activity

[0394]

[0395]

[0396] The test results show that by replacing the methoxyphenyl group of comparative compound 2 (AOH-1996) with a specific bicyclic ring, or further replacing the O oxygen atom between the two benzene rings with an S atom, the anti-tumor (HL-60) activity of the compound is significantly enhanced, such as the HL-60 inhibitory activity of compound 8 (MTB-1956) is 3 times that of comparative compound 2 (AOH-1996), and the HL-60 inhibitory activity of compound 44, compound 90, and compound 93 is about 9 times that of comparative compound 2 (AOH-1996). In comparative compound 1 (MTB-1962), after replacing the methoxyphenyl group of comparative compound 2 (AOH-1996) with a bicyclic ring of benzothiazole, the activity of comparative compound 1 is significantly lower, and its activity is 11 times lower than that of comparative compound 2 (AOH-1996). The amino group is usually easily oxidized by P450, and the substitution of fluorine can improve its biological stability. 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.

[0397] Table 4 Compounds inhibiting the activity of NCI-H522

[0398]

[0399]

[0400] The test results show that the anti-tumor (NCI-H522) activity of the compound is significantly enhanced by replacing the methoxyphenyl group of AOH-1996 with a specific bicyclic ring, such as compound 8 (MTB-1956). However, the activity of the comparative compound 1 is lower after replacing the methoxyphenyl group of AOH-1996 with a benzothiazole bicyclic ring.

[0401] Broad-spectrum anti-tumor activity test:

[0402] 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):

[0403]

[0404]

[0405] The test results show that the inhibitory activity of compound 44 (MTB-1970) and compound 99 and their methyl deuterated compounds 93 (MTB-1993) and compound 100 (MTB-2006) on all tumor cells tested is 5-10 times that of AOH-1996, and there is no obvious cytotoxicity to normal human peripheral blood mononuclear cells at the highest dose tested, 10 μM concentration. It shows great development potential. The anti-tumor activity of compound 100 (MTB-2006) after the hydrogen at the amino position of compound 93 (MTB-1993) is replaced by fluorine is equivalent to that of compound 93 (MTB-1993).

[0406] Pharmacokinetic studies

[0407] Pharmacokinetic study of single oral administration of compound 2 (AOH-1996), compound 100 (MTB-2006) and compound 99 in female CB-17 SCID (B-Ces1c KO) mice

[0408] 1. Research methods and experimental design

[0409] 1.1 Test system

[0410]

[0411] 1.2 Experimental Design

[0412] 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.

[0413] 2. Sample collection

[0414] 2.1. Drug delivery preparations

[0415] After each preparation was prepared, two portions of clear dosing solution (upper and bottom layers) or three portions of suspension (upper, middle and bottom layers) were collected and analyzed by HPLC-UV to investigate the accuracy of the preparation concentration.

[0416] 2.2 Sample collection and processing

[0417] Plasma samples were collected at 0.167, 0.333, 0.5, 1, 2, 4, 6, and 12 h after administration.

[0418] Whole blood collection and plasma sample preparation: At each time point, blood samples were collected from the saphenous vein of each animal (approximately 0.02 μg / mL at each time point) using pre-cooled commercial blood collection tubes (containing EDTA-K2 as an anticoagulant).

[0419] mL) whole blood was collected, and stabilizer was added within 10 seconds at a ratio of whole blood: 400mM DDV. (v:v, 19:1) and placed on wet ice until centrifugation. Each blood collection will be placed on wet ice before centrifugation. Blood samples will be centrifuged at approximately 4°C, 3,200g for 10 minutes to prepare plasma samples. Then they were transferred to low-absorption tubes, quickly frozen on dry ice, and stored at -60°C or lower until LC-MS / MS. Sample processing was performed under wet ice conditions.

[0420] 3. Sample analysis

[0421] 3.1. Drug delivery preparations

[0422] The concentration of the drug preparation was determined by HPLC-UV method, and the calibration curve contained at least 6 concentration levels. Each sample was analyzed twice in parallel, and the actual measured concentration should be within 80 to 120% of the theoretical preparation concentration, otherwise the pharmacokinetic parameters should be calculated based on the actual dose.

[0423] 3.2 Bioanalytical method development and sample analysis

[0424] Under Non-GLP conditions, an LC-MS / MS quantitative determination method for the test compounds in biological matrices was established.

[0425] Table 6. Pharmacokinetic data of AOH-1996, compound 99 and MTB-2006

[0426] AOH-1996 Compound 99 MTB-2006 AUC 0-last (ng.h / mL) 162 345 457 Cmax(ng / mL) 54.3 68.6 98.5 <![CDATA[T 1 / 2 (h)]]> 2.99 3.21 3.73

[0427] The results of the pharmacokinetic test (Table 6) show that compound 99 and compound 100 (MTB-2006) have better biostability. The deuterated 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 with compound 99. max The half-life and duration of MTB-2006 were significantly improved compared with those of AOH-1996.

[0428] From the above test results, it can be seen that the inhibitory activity of the exemplary compounds on various tumor cells in vitro is 5-10 times that of the comparative compound 2 (AOH-1996), and its biological stability and drug exposure are also significantly improved, which has greater development potential and clinical value.

[0429] The above is an exemplary description of the specific implementation of the present invention. However, the present invention is not limited to the above specific implementation. Any modification, equivalent replacement, etc. made within the spirit and principle 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 Both of them are hydrogen 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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