ATR inhibitor as well as preparation method and application thereof

Abstract

The invention discloses an ATR inhibitor as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The ATR inhibitor prepared by the preparation method disclosed by the invention can be used for effectively inhibiting ATR activity and reducing dependence and division and proliferation of tumor cells, and has the potential of researching and developing tumor targeted therapy.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly relates to an ATR inhibitor, a preparation method thereof, and an application thereof in cancer treatment. Background Art

[0002] Since the 1990s, a series of protein families with significantly different structures from typical serine / threonine protein kinases (STKs) have been gradually discovered. They have a relatively large molecular weight (between 280 and 470 kDa), and their domains are more similar in sequence to the phosphatidylinositol 3-kinase (PI3K) family (about 20-25% identical). Therefore, they are called phosphatidylinositol-3-kinase-related kinases (PIKK) family. [1] The PIKK family is ubiquitous in all eukaryotes and mainly includes ATM, ATR (ataxia telangiectasia and Rad3-related protein), DNA-PKcs (DNA-dependent protein kinase catalytic subunit), mTOR (mammalian target of rapamycin), SMG1 (suppressor of morphogenesis in genitalia 1), and TRRAP (transformation / transcription-associated protein) without protein kinase activity, etc. Members of the PIKK family are involved in a wide variety of cell activity regulations. For example, ATM, ATR, and DNA-PKcs are involved in the activation of the DNA damage response pathway, mTOR controls cell growth, proliferation, and metabolism, and SMG1 and TRRAP play key roles in the decay of mRNA and the epigenetic regulation of transcription, respectively. Many studies have shown that PIKK family kinases are closely related to the occurrence and development of tumors, making these kinases potential targets in cancer treatment.

[0003] Tens of thousands of DNA damages occur in human cells every day. The accumulation of DNA damages can lead to genomic instability and promote the occurrence of cancer. Therefore, cells have evolved extensive and complex DNA damage response (DDR) pathways to identify and process these specific types of DNA damages. ATR is also a key kinase in the DDR regulatory network. Different from ATM that responds to DNA double-strand break signals (DSBs), ATR is specifically activated by a wider range of single-stranded DNA damages (ssDNAs). There are many reasons for the formation of ssDNA, such as DNA replication stalling, resection of the damaged ends of DNA DSBs, DNA double-strand cross-linking, base adducts, and inhibition of DNA polymerase. In addition, ATR can be directly induced by ultraviolet irradiation, while ionizing radiation indirectly activates ATR by converting DSB damages into ssDNA through the activation of ATM. The functions of ATR are mainly reflected in: (1) regulating the cell cycle to inhibit the replication of damaged DNA. ATR is mainly activated during the S phase and the subsequent G2 phase when DNA is rapidly synthesized in cells to participate in DDR functions. Subsequently, CHK1 is phosphorylated and mediates the degradation of CDC25A, resulting in the inhibition of CDK1, which activates the S-phase or G2 / M-phase cell cycle checkpoints, delaying the time for DNA repair. In addition, phosphorylated CHK1 can also activate the G1 / S cell cycle checkpoint by mediating the degradation of CDC25C in response to DNA damages induced by ultraviolet light. (2) Protection and restart of replication forks. ATR phosphorylates downstream proteins such as SMARCAL1, inhibits kinases such as RNF4 or PLK1, etc., reducing the formation of DSBs mediated by the SLX4 structure-specific endonuclease and avoiding the collapse of replication forks. ATR also stabilizes the localization of RPA when replication forks stall by phosphorylating WRN. (3) Inhibiting mitotic abnormalities and catastrophes.

[0004] The decrease in ATR activity leads to defects in mitotic functions in cells, such as chromosome breakage, anaphase bridges, DNA micronuclei, etc. Therefore, the activation of ATR prevents cells with damaged DNA from prematurely entering the mitotic phase and avoids cell death caused by mitotic catastrophe.

[0005] The activation of ATR plays an important role in DNA single-strand damage repair, promotes the initiation of stalled replication forks and restores the DNA replication process, and maintains genomic stability by activating the cellular S or G2 / M cycle checkpoints. Therefore, targeting ATR has become a potential therapeutic target in cancer treatment. ATR activation mutations can lead to a rare autosomal recessive genetic disorder, Seckel syndrome (SS), which is characterized by short stature, skull deformities, and eye lesions. Different from other genetic diseases caused by the absence of DDR protein expression, such as A-T (ataxia-telangiectasia). ATR activity is essential in all proliferating cells, but low levels of ATR activity are sufficient to maintain the viability of highly proliferating adult tissues, indicating that pharmacological inhibition of ATR has less impact on normal tissue homeostasis. Tumor cells are more sensitive to ATR inhibition due to differences in DDR. Defects in one or more DDR pathways, a significant increase in replication stress, and an elevated level of endogenous DNA damage are the three main differences between cancer cells and normal cells. Therefore, the cancer DNA damage response pathway is a potential source of drug targets with fewer side effects. Replication stress (RS) is the slowdown and stalling of DNA replication forks. Factors such as replication errors, nucleotide deficiencies, DNA structural overlaps, and conflicts between replication and transcription functions can all lead to the emergence of replication stress. Cancer cells may be more dependent on ATR activation to reduce the impact of replication stress and achieve rapid proliferation. Finally, various types of ATM mutations and defects have been found in multiple tumors, while mutations and defects in ATR rarely occur in tumor cells, further verifying the dependence of tumors on the ATR pathway. Overall, although ATR is crucial for the development and survival of organisms, its inhibition may have less impact on normal cell survival and organismal homeostasis. Tumor cells are more sensitive to ATR inhibition due to replication stress and synthetic lethality and are more likely to become tumor-dependent. Therefore, targeting ATR is a potential tumor treatment strategy. Summary of the Invention

[0006] To solve the above problems, the present invention provides an ATR inhibitor, its preparation method, and its application in the treatment of genetic defects and tumors.

[0007] To achieve the above object, the present invention is realized through the following technical solutions:

[0008] An ATR inhibitor, which is a compound having the following structural formula I:

[0009] Wherein A and / or B is a nitrogen-containing heterocyclic group or heteroaryl group, or a nitrogen- and oxygen-containing heterocyclic group or heteroaryl group, or a nitrogen-, oxygen-, and fluorine-containing heterocyclic group or heteroaryl group, or a nitrogen-, oxygen-, fluorine-, and sulfur-containing heterocyclic group or heteroaryl group.

[0010] Further, the A and B rings in general formula I are R1 is any one or more of a hydrogen atom, a halogen, an alkyl group, and a substituted alkyl group.

[0011] Further, the ATR inhibitor includes the following structure:

[0012]

[0013] The present invention also discloses a preparation method of an ATR inhibitor, which is characterized by comprising the following steps:

[0014] (1) Using 2,4-dichloropurine as a starting material, reacting with dihydropyran (DHP) under acidic conditions to obtain intermediate 1, and then selectively reacting the chlorine at the 2-position of pyrimidine with (R)-3-methylmorpholine under the alkaline condition of K2CO3 to obtain a single-substituted intermediate 2;

[0015] (2) Under the conditions of anhydrous, N2 protection and -78 °C, the H on the imidazole of intermediate 2 is removed by n-butyllithium, forming a relatively stable C anion with tetramethylethylenediamine in the solvent, and then DMF is added dropwise, and finally an aldehyde intermediate 3 is formed. Intermediate 3 is then successively reduced by sodium borohydride and the tetrahydropyran (THP) is removed under acidic conditions to obtain intermediate 5;

[0016] (3) 1,2-dibromoethane is used as a ring-closing fragment group, and nucleophilic substitution reactions occur with a hydroxyl group and an amino group on both sides under alkaline conditions to obtain a tricyclic system intermediate 6 of morpholinoimidazopyrimidine; Intermediate 6 undergoes a Suzuki-Miyaura coupling reaction under the catalysis of Pd(Ph3P)4 to obtain the final product compound C01, compound C05-26, and the reaction formula is Scheme 1;

[0017] The specific reaction formula of Scheme 1 is:

[0018]

[0019] (4) The intermediate 6 is converted to the nitro compound intermediate 19 through Buchwald-Hartwig coupling reaction; the intermediate 19 is reduced to the amino-substituted benzene ring / heterocyclic derivative intermediate 20, and the intermediate 20 directly reacts with cyanogen bromide (BrCN) to obtain compounds C27 and C37; the intermediate 20 reacts with N,N'-carbonyldiimidazole (CDI) to obtain compound C28; when pyridine is used as the solvent and base, the intermediate 20 reacts with methyl isothiocyanate to obtain the methylthiourea intermediate 21; the intermediate 21 uses 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) as a condensing agent and undergoes ring closure under heating conditions to obtain the N-methylbenzimidazole compounds C29-36; the reaction formula is Scheme 4.

[0020] The specific reaction formula of Scheme 4 is as follows:

[0021]

[0022] The present invention provides another preparation method of an ATR inhibitor, and the steps are as follows:

[0023] (1) Using 2,6-dichloro-3-nitropyridin-4-amine as the starting material, first reacting with (R)-3-methylmorpholine under heating in the presence of an organic base to obtain the singly substituted intermediate 8 at the 2-position in a relatively high yield;

[0024] (2) Subsequently, the nitro group is reduced under the condition of Fe / HCl to obtain the aniline intermediate 9; the intermediate 9 and triethyl orthoformate undergo ring closure to obtain the imidazopyridine intermediate 10; the intermediate 10 reacts with DHP under acidic conditions to obtain the intermediate 11;

[0025] (3) Under the conditions of anhydrous, N2 protection and -78 °C, the H on the imidazole of the intermediate 11 is removed by lithium diisopropylamide (LDA) to form a relatively stable C anion with tetramethylethylenediamine in the solvent, and then DMF is added dropwise, and finally the aldehyde intermediate 12 is formed. The intermediate 12 is then reduced by sodium borohydride in sequence to obtain the intermediate 13. The intermediate 13 removes THP under acidic conditions to obtain the intermediate 14. The intermediate 14 undergoes ring closure under the conditions of potassium tert-butoxide and an aqueous solution of tetrabutylammonium bromide (TBAB) to obtain the intermediate 15. The intermediate 15 undergoes Suzuki-Miyaura coupling reaction under the catalysis of Pd(Ph3P)4 to obtain the compound C02; the reaction formula is Scheme 2, and the specific reaction formula of Scheme 2 is as follows:

[0026]

[0027] (4) The intermediate 11 is deprotonated in the presence of lithium diisopropylamide (LDA) and directly reacts with 1,3-dibromopropane or 1,4-dibromobutane to obtain the intermediate 16 with a single substituent. After removing THP, the imidazopyridine nucleus with exposed NH is obtained. Ring closure under basic conditions gives the intermediate 18, and the intermediate 18 undergoes a Suzuki-Miyaura coupling reaction under the catalysis of Pd(Ph3P)4 to obtain compounds C03 and C04. The reaction formula is Scheme 3, and the specific reaction of Scheme 3 is as follows:

[0028]

[0029] The present invention also protects a compound represented by the general formula I, or its stereoisomer, or its pharmaceutically acceptable salt, which is used for treating or improving gene defects and / or tumors.

[0030] Furthermore, the gene defect is the ataxia telangiectasia mutated gene, and the tumors include gastric cancer solid tumors, colorectal cancer solid tumors, head and neck squamous cell carcinomas, small cell lung cancers with DDR mutations, ovarian cancers, leukemias, B-cell lymphomas, and triple-negative breast cancers with HR mutations.

[0031] The present invention also protects a pharmaceutical composition containing the compound represented by the general formula I, or its stereoisomer, or its pharmaceutically acceptable salt.

[0032] Furthermore, the pharmaceutical composition further includes a pharmaceutically acceptable carrier, excipient, auxiliary material, and vehicle.

[0033] An ATR inhibitor of the present invention, its preparation method and application have the beneficial effects that: the ATR inhibitor obtained by the preparation method of the present invention can effectively inhibit ATR activity, reduce the dependence and division and proliferation of tumor cells, and has the potential for developing tumor-targeted therapies. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0035] Figure 1 is the kinase selectivity of compound C27 of the present invention.

[0036] Figure 2 is a schematic diagram of the in vitro antitumor activity of compound C27 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] To enable those skilled in the art to better understand the solution of the present invention, the technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0038] Example 1

[0039] A method for preparing an ATR inhibitor intermediate, comprising the following steps:

[0040] (1) 2,4-Dichloropurine (20 g, 105.8 mmol) and p-toluenesulfonic acid (0.9 g, 5 mmol) were dissolved in 200 ml of tetrahydrofuran, and DHP (26.7 g, 317.5 mmol) was gradually added dropwise with stirring. After the addition was completed within 5 minutes, stirring was continued at room temperature for about 30 minutes, and the reaction system changed from turbid to yellow-brown and clear. TLC plate monitoring showed that the raw materials had reacted completely. About 200 ml of ammonia water and 100 ml of ethyl acetate were added, and the organic phases were extracted and combined three times, and then concentrated under reduced pressure to obtain a yellow viscous liquid. Appropriate amount of petroleum ether was added for pulping to obtain a white solid, which was filtered and washed twice with petroleum ether, and then dried to obtain 25.7 g of white intermediate 1, with a yield of 89%;

[0041] (2) Intermediate 1 (20 g, 73.2 mmol) and K2CO3 (20.2 g, 146.4 mmol) were placed in a 250-ml eggplant-shaped flask, and 150 ml of acetonitrile was added to disperse them fully. (R)-3-Methylmorpholine (14.8 g, 146.4 mmol) was added dropwise with stirring, and stirring was continued at room temperature overnight. The next day, TLC plate monitoring showed that the raw materials had reacted completely. The potassium carbonate solid particles were removed by suction filtration through diatomaceous earth, and the filtrate was directly concentrated under reduced pressure to obtain a pale yellow viscous liquid. Appropriate amount of ether was added for pulping, and then suction filtered and dried to obtain 20.5 g of white intermediate 2, with a yield of 83%;

[0042] (3) Intermediate 2 (20 g, 59 mmol) and tetramethylethylenediamine (10.3 g, 88.8 mmol) were dissolved in 200 ml of anhydrous THF, and the temperature was lowered to -78 °C in a cold well. Subsequently, under N2 protection, 2.5 M n-butyllithium n-BuLi (35.5 ml, 88.8 mmol) was slowly added dropwise. After 30 minutes, the addition was complete, and the mixture was stirred at -78 °C for 1 h. Fumarate (DMF) (6.48 g, 88.8 mmol) was then slowly added at low temperature. After the addition was complete, the mixture was stirred for 1 h, and finally the reaction system was allowed to warm to room temperature naturally and stirred for about 2 h. TLC monitoring showed that the raw materials had basically reacted completely. The reaction was quenched by slowly adding about 200 ml of water in an ice bath. About 100 ml of ethyl acetate was added again, and the organic phases were extracted and combined three times. The solvent was concentrated under reduced pressure and purified by flash column chromatography (petroleum ether / ethyl acetate = 9 / 1) to obtain 18.6 g of pale yellow intermediate 3 with a yield of 86%;

[0043] (4) Intermediate 3 (15 g, 41 mmol) was dissolved in 100 ml of methanol, and solid sodium borohydride NaBH4 (1.87 g, 49 mmol) was slowly added in an ice bath, and gas bubbles gradually appeared in the reaction system. Subsequently, the temperature was raised to room temperature and stirred for about 2 h. TLC monitoring showed that the raw materials had reacted completely. The solvent methanol was removed by concentration under reduced pressure. A total of 200 ml of ethyl acetate / water (volume ratio 1 / 1) was added for extraction, and the organic phases collected three times were combined and dried by rotation to obtain 14.3 g of white intermediate 4a, which did not require further purification, with a yield of 95%;

[0044] (5) Intermediate 2 (3 g, 8.9 mmol) and tetramethylethylenediamine (1.55 g, 13.3 mmol) were dissolved in 30 ml of anhydrous THF, and the temperature was lowered to -78 °C in a cold well. Subsequently, under N2 protection, 2.5 M n-BuLi (5.3 ml, 13.3 mmol) was slowly added dropwise. After 5 minutes, the addition was complete, and the mixture was stirred at -78 °C for 1 h. Acetone (0.77 g, 13.3 mmol) was then slowly added at low temperature. After the addition was complete, the mixture was stirred for 1 h, and finally the reaction system was allowed to warm to room temperature naturally and stirred for about 2 h. TLC monitoring showed that the raw materials had basically reacted completely. The reaction was quenched by slowly adding about 30 ml of water in an ice bath. About 30 ml of ethyl acetate was added again, and the organic phases were extracted and combined three times. The solvent was concentrated under reduced pressure and purified by flash column chromatography (petroleum ether / ethyl acetate = 8 / 1) to obtain 1.62 g of pale yellow intermediate 4b with a yield of 46%;

[0045] (6) The intermediate 4a (14 g, 38 mmol) was dissolved in 100 ml of methanol. Approximately 5 ml of 3M aqueous HCl solution was added dropwise at room temperature, and then the mixture was heated to 50 °C and stirred for 3 h. TLC monitoring showed that the raw materials had reacted completely. After most of the methanol was removed by concentration under reduced pressure, 1M aqueous NaOH solution was added dropwise to the reaction system to adjust the pH to about 8, and a large amount of white solid appeared. The solid was filtered by suction and washed with water to obtain the filter cake layer, which was dried to obtain 8.85 g of white intermediate 5a and 5a. Without further purification, the yield of intermediate 5a was 82%; the yield of intermediate 5b was 85%;

[0046] (7) Intermediate 5a (8 g, 28.2 mmol), 1,2-dibromoethane (15.9 g, 84.6 mmol), TBAB (2.73 g, 8.5 mmol) and potassium tert-butoxide (6.32 g, 56.4 mmol) were dissolved in 80 ml of water. 1,2-Dibromoethane served as the ring-closing fragment group. The mixture was heated to 90 °C and stirred for about 10 h in a sealed pressure-resistant tube. TLC monitoring showed that the raw materials had reacted completely. The water was directly removed by concentration under reduced pressure and purified by flash column chromatography (dichloromethane / methanol = 49 / 1) to obtain 6.4 g of intermediate 6a. Beige solid, yield 73%; The synthesis method of intermediate 6b was the same as that of intermediate 6a, and intermediate 5b was used instead of intermediate 5a;

[0047] The specific reaction formula is as follows:

[0048]

[0049] (8) Intermediate 6a (300 mg, 0.97 mmol), 2-nitroaniline (44 mg, 1.16 mmol), Pd2(dba)3 (89 mg, 0.10 mmol), Xantphos (112 mg, 0.19 mmol) and t-BuOK (217 mg, 1.94 mmol) were dissolved in 15 ml of anhydrous 1,4-dioxane. The mixture was stirred at 100 °C overnight under a N2 atmosphere. The next day, TLC monitoring showed that the raw materials had reacted completely. After it was cooled to room temperature, the insoluble catalysts, ligands and organic bases were removed by suction filtration through diatomaceous earth. After the filtrate was concentrated under reduced pressure, it was purified by flash column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain 342 mg of intermediate 19a. Yellow solid, yield 86%;

[0050] The synthesis methods of intermediates 19b - 19h were the same as that of 19a, and the corresponding 2-nitroaniline derivatives were used instead of 2-nitroaniline. After TLC monitoring showed that the reaction was basically complete, the clear black filtrate was first obtained by suction filtration. After concentration under reduced pressure, it was preliminarily purified by flash column chromatography;

[0051] (9) Intermediate 19a (200 mg) was dissolved in 10 ml of methanol, about 20 mg of Pd / C solid was added, and the mixture was stirred at room temperature for about 6 h under a H2 atmosphere. After monitoring by TLC that the raw material reaction was complete, it was filtered through diatomaceous earth to obtain a light yellow clear solution, which was concentrated under reduced pressure and purified by flash column chromatography (dichloromethane / methanol = 97 / 3) to obtain 247 mg of Intermediate 20a. A light gray solid, with a yield of 89%;

[0052] The synthesis method of Intermediate 20b is the same as that of Intermediate 20a, just replace 19a with Intermediate 19b. A light yellow solid, with the total yield of the two-step reaction being 65%;

[0053] The synthesis method of Intermediate 20e is the same as that of Intermediate 20a, just replace 19a with Intermediate 19e. A light yellow solid, with the total yield of the two-step reaction being 68%;

[0054] The synthesis method of Intermediate 20e is the same as that of Intermediate 20a, just replace 19a with Intermediate 19f. A light yellow solid, with the total yield of the two-step reaction being 64%;

[0055] The synthesis method of Intermediate 20h is the same as that of Intermediate 20a, just replace 19a with Intermediate 19h. A light yellow solid, with the total yield of the two-step reaction being 62%.

[0056] Example 2

[0057] A preparation method of an ATR inhibitor intermediate, comprising the following steps:

[0058] (1) 4-Bromo-6-trifluoromethyl-1H-indole (300 mg, 1.1 mmol), bis(pinacolato)diboron (577 mg, 2.2 mmol), PdCl2(dppf) (83 g, 0.1 mmol) and AcOK (223 mg, 2.2 mmol) were placed in a 50 ml three-necked round-bottom flask, about 20 ml of anhydrous dioxane was added as a solvent, and the mixture was heated to 100 °C and stirred overnight under a N2 atmosphere. The next day, TLC was used to monitor that the reaction was basically complete. After it was cooled to room temperature, it was filtered through diatomaceous earth to collect the filtrate, and the black crude product Intermediate 7a was obtained by concentration under reduced pressure and could be directly used for the next step without further purification;

[0059] The synthesis method of Intermediate 7b is the same as that of Intermediate 7a, just replace 4-bromo-6-trifluoromethyl-1H-indole with 4-bromo-6-methoxy-1H-indole. A black crude product, which could be directly used for the next step without further purification;

[0060] The synthesis method of intermediate 7c is the same as that of intermediate 7a, except that 4-bromo-6-(methoxycarbonyl)-1H-indole is used instead of 4-bromo-6-(trifluoromethyl)-1H-indole. It is purified by flash column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain a white solid with a yield of 76%.

[0061] The specific reaction formula is as follows:

[0062]

[0063] Example 3

[0064] A preparation method of an intermediate of an ATR inhibitor, comprising the following steps:

[0065] (1) 2,6-Dichloro-3-nitropyridin-4-amine (10 g, 48 mmol) and Et3N (9.7 g, 96 mmol) are dissolved in 100 ml of DMA, and (R)-3-methylmorpholine (9.7 g, 96 mmol) is added dropwise at room temperature. After the addition is complete, the temperature is raised to 80 °C and stirred for about 5 h. Monitored by TLC plate spotting, the raw materials have reacted completely. After it is cooled to room temperature, 400 ml of a mixed solvent of ethyl acetate / water (volume ratio 1 / 3) is added. After extraction, the organic phases collected three times are combined, dried over anhydrous Na2SO4 solid, concentrated under reduced pressure and purified by flash column chromatography (ethyl acetate / petroleum ether = 4 / 1) to obtain 11.7 g of intermediate 8. It is a bright yellow solid with a yield of 89%;

[0066] (2) Intermediate 8 (10 g, 37 mmol) and reduced iron powder (8.2 g, 148 mmol) are placed in a 250 ml eggplant-shaped flask, and 100 ml of a mixed solvent of methanol / water (volume ratio 9 / 1) is added. 5 ml of concentrated hydrochloric acid is added dropwise, and then heated to about 90 °C for reflux. After stirring for about 5 h, TLC plate spotting monitors that the raw materials have completely reacted. The large amount of insoluble Fe powder is removed by hot filtration with diatomaceous earth to obtain a clear yellow filtrate. Concentrate under reduced pressure, add an appropriate amount of water for pulping, filter again and dry the filter cake layer to obtain 6.9 g of light yellow intermediate 9 with a yield of 78%;

[0067] (3) Place intermediate 9 (6.5 g, 26.7 mmol) in a 250 ml eggplant-shaped flask, and successively add 25 ml of triethyl orthoformate and 25 ml of acetic anhydride. Then heat to 90 °C and stir for 2 h. After monitoring by TLC that the raw materials have reacted completely, cool the above-mentioned black reaction solution to room temperature, and white solid gradually precipitates. Filter by suction to obtain the solid in the filter cake layer, add 50 ml of 5M NaOH solid thereto, then heat to 85 °C and stir for 1 h. After monitoring by TLC that the raw materials have reacted completely, cool the reaction solution to room temperature, slowly add 3N dilute hydrochloric acid aqueous solution and adjust the pH to about 7, and a large amount of solid gradually appears. Filter by suction again, wash with water and dry the filter cake layer to obtain 4.4 g of intermediate 10 without further purification. White solid, yield 65%;

[0068] (4) Dissolve 2,4-dichloropurine (20 g, 105.8 mmol) and p-toluenesulfonic acid (0.9 g, 5 mmol) in 200 ml of tetrahydrofuran, and gradually add dropwise DHP (26.7 g, 317.5 mmol) under stirring at 50 °C. After adding dropwise within 5 minutes, continue to stir at 50 °C for about 3 h, and the reaction system changes from turbid to yellow-brown and clear. After monitoring by TLC that the raw materials have reacted completely, add about 200 ml of ammonia water and 100 ml of ethyl acetate, extract and combine the organic phases three times, and concentrate under reduced pressure to obtain a yellow viscous liquid. Add an appropriate amount of petroleum ether to slurry to obtain a white solid, filter and wash twice with petroleum ether, and dry to obtain white intermediate 11, yield 86%;

[0069] (5) Dissolve intermediate 11 (4.5 g, 13.4 mmol) in 50 ml of anhydrous THF, and cool to -78 °C in a cold well. Subsequently, under N2 protection, slowly add dropwise 2M LDA (10 ml, 20 mmol). After adding dropwise for 10 minutes, keep stirring at -78 °C for 1 h. Then slowly add DMF (1.46 g, 20 mmol) at low temperature. After adding dropwise, continue to stir for 1 h, and finally let the reaction system rise to room temperature naturally and stir for about 2 h. After monitoring by TLC that the raw materials have basically reacted completely, slowly add about 50 ml of water to quench the reaction in an ice bath. Add about 50 ml of ethyl acetate again, extract and combine the organic phases three times, concentrate under reduced pressure, and purify by flash column chromatography (petroleum ether / ethyl acetate = 8 / 1) to obtain 3.1 g of pale yellow intermediate 12, yield 63%;

[0070] (6) Dissolve intermediate 12 (15 g, 41 mmol) in 100 ml of methanol. Slowly add solid NaBH4 (1.87 g, 49 mmol) under an ice bath, and bubbles gradually appear in the reaction system. Subsequently, warm the temperature to room temperature and stir for about 2 h. Monitor by TLC plate that the raw materials have reacted completely, and concentrate under reduced pressure to remove the solvent methanol. Add a total of 200 ml of ethyl acetate / water (volume ratio 1 / 1) for extraction. Combine the organic phases collected three times and rotary evaporate to obtain white intermediate 13, without further purification, and the yield is 91%;

[0071] (7) Dissolve intermediate 13 (14 g, 38 mmol) in 100 ml of methanol. Dropwise add about 5 ml of 3M HCl aqueous solution at room temperature, then heat to 50 °C and stir for 3 h. Monitor by TLC plate that the raw materials have reacted completely. After concentrating under reduced pressure to remove most of the methanol, add 1M NaOH aqueous solution dropwise to the reaction system to adjust the pH to about 8, and a large amount of white solid appears. Filter by suction and wash with water to obtain the filter cake layer, and dry to obtain white intermediate 14, without further purification, and the yield is 85%;

[0072] (8) Dissolve intermediate 14 (8 g, 28.2 mmol), 1,2-dibromoethane (15.9 g, 84.6 mmol), TBAB (2.73 g, 8.5 mmol) and potassium tert-butoxide (6.32 g, 56.4 mmol) in 80 ml of water. Heat to 90 °C in a sealed pressure-resistant tube and stir for about 10 h. Monitor by TLC plate that the raw materials have reacted completely. Directly concentrate under reduced pressure to remove water, and purify by flash column chromatography (dichloromethane / methanol = 49 / 1) to obtain a white solid, namely intermediate 15, and the yield is 69%; The specific reaction equations for steps (1) - (8) are as follows:

[0073]

[0074] (9) The synthesis method of intermediate 16a is the same as that of intermediate 12, just replace DMF with 1,3-dibromopropane. Light yellow solid, and the yield is 43%; The synthesis method of intermediate 16b is the same as that of intermediate 12, just replace DMF with 1,4-dibromobutane. Light yellow solid, and the yield is 62%;

[0075] (10) The synthesis method of intermediate 17a is the same as that of intermediate 14, just replace intermediate 13 with intermediate 16a. White solid, and the yield is 83%; The synthesis method of intermediate 17b is the same as that of intermediate 14, just replace intermediate 13 with intermediate 16b. White solid, and the yield is 86%;

[0076] (11) Intermediate 17a (400 mg, 1.07 mmol) was dissolved in 10 ml of acetonitrile, Cs2CO3 (698 mg, 2.14 mmol) was added, and the mixture was heated to 80 °C and stirred overnight. The next day, TLC was used to monitor the reaction progress, and it was found that the raw material had completely reacted. The Cs2CO3 solid was removed by filtration, the filtrate was concentrated under reduced pressure, and purified by flash column chromatography (petroleum ether / ethyl acetate = 3 / 2) to obtain 119 mg of intermediate 18a. It was a white solid with a yield of 38%. The synthesis method of intermediate 18b was the same as that of intermediate 18a, except that intermediate 17b was used instead of intermediate 17a. It was a white solid with a yield of 51%. The specific reaction equations for steps (9)-(11) are as follows:

[0077]

[0078] Example 4

[0079] A method for preparing an ATR inhibitor C01 compound, comprising the following steps:

[0080] Intermediate 6a (100 mg, 0.32 mmol) obtained in Example 1, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine (98 mg, 0.39 mmol), Pd(Ph3P)4 (37 mg, 0.03 mmol) and K2CO3 (89 mg, 0.64 mmol) were placed in a 25 ml three-necked flask, and 8 ml of a mixed solvent of 1,4-dioxane / water (volume ratio 7 / 1) was added. Under a nitrogen atmosphere, the above mixture was heated to 100 °C and stirred overnight. The next day, TLC was used to monitor the reaction progress, and it was found that the raw materials had completely reacted. After the reaction was cooled to room temperature, the filtrate was obtained by filtration through diatomaceous earth and concentrated to obtain a black viscous substance, which was purified by a flash preparative column machine (methylene chloride / methanol = 98 / 2) to obtain 100 mg of white compound C01 with a yield of 83%. The NMR data of compound C01 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.78 (s, 1H), 9.14 (s, 1H), 8.81 (s, 1H), 7.77–7.66 (m, 1H), 7.44 (d, J = 2.9 Hz, 1H), 5.46 (s, 1H), 5.06 (s, 1H), 4.95 (s, 2H), 4.26 (d, J = 5.3 Hz, 2H), 4.17 (d, J = 5.1 Hz, 2H), 4.07–3.97 (m, 1H), 3.81 (d, J = 11.4 Hz, 1H), 3.76–3.69 (m, 1H), 3.58 (t, J = 11.6 Hz, 1H), 3.51–3.42 (m, 1H), 1.35 (d, J = 6.8 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 157.09, 152.34, 151.69, 144.97, 138.84, 134.94, 130.47, 130.26, 124.96, 117.24, 102.99, 70.37, 66.47, 64.43, 63.30, 41.10, 14.28. HRMS (ESI), m / z: 392.1839 [M+H+].

[0081] Example 5

[0082] A preparation method of an ATR inhibitor C02 compound. The synthesis method of compound C02 is the same as that of C01 in Example 4, replacing intermediate 6a with intermediate 15 obtained in Example 3 and replacing 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 6-azaindole-4-borate. White solid, yield 75%. The NMR data of compound C02 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.72 (s, 1H), 8.74 (s, 1H), 8.66 (s, 1H), 7.68 (s, 1H), 7.47 (s, 1H), 7.05 (d, J = 2.9 Hz, 1H), 5.46 (q, J = 7.1 Hz, 1H), 4.97 (s, 2H), 4.82 (d, J = 13.4 Hz, 1H), 4.25 (d, J = 4.0 Hz, 2H), 4.18 (d, J = 5.0 Hz, 2H), 3.98 (dd, J = 11.3, 3.4 Hz, 1H), 3.80–3.72 (m, 2H), 3.58 (dt, J = 11.6, 6.0 Hz, 1H), 3.40 (dd, J = 12.3, 3.7 Hz, 1H), 1.25 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 149.38, 146.36, 145.54, 141.02, 137.01, 133.61, 133.54, 129.78, 127.62, 125.85, 101.29, 95.62, 70.66, 66.73, 64.49, 63.33, 47.65, 42.11, 13.60. HRMS (ESI), m / z: 391.1887 [M+H+].

[0083] Example 6

[0084] A preparation method of an ATR inhibitor C03 compound. The synthesis method of compound C03 is the same as that of C01 in Example 4, and intermediate 18a obtained in Example 3 is used to replace intermediate 6a. It is a white solid with a yield of 85%. The NMR data of compound C03 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 11.73 (s, 1H), 8.74 (s, 1H), 8.64 (s, 1H), 7.69 (d, J = 3.1 Hz, 1H), 7.43 (s, 1H), 7.04 (d, J = 2.9 Hz, 1H), 5.50 (d, J = 7.2 Hz, 1H), 4.80 (d, J = 13.4 Hz, 1H), 4.20–4.11 (m, 2H), 3.98 (dd, J = 11.2, 3.4 Hz, 1H), 3.80–3.68 (m, 2H), 3.59 (td, J = 11.6, 2.8 Hz, 1H), 3.33 (s, 1H), 2.96 (td, J = 7.4, 2.8 Hz, 2H), 2.64 (p, J = 7.4 Hz, 2H), 1.24 (d, J = 6.7 Hz, 3H). HRMS (ESI), m / z: 375.1941 [M+H+].

[0085] Example 7

[0086] A preparation method of an ATR inhibitor C04 compound. The synthesis method of compound C04 is the same as that of C01 in Example 4, and intermediate 18b obtained in Example 3 is used to replace intermediate 6a. It is a white solid with a yield of 78%. The NMR data of compound C04 are as follows: 1 H NMR(400MHz,DMSO-d6)δ12.17(s,1H),8.85(s,1H),8.68(s,1H),7.89(d,J=3.0Hz,1H),7.46(s,1H),7.17(d,J=3.0Hz,1H),5.49(d,J=7.2Hz,1H),4.81(d,J=13.4Hz,1H),4.21–4.08(m,2H),3.98(dd,J=11.3,3.3Hz,1H),3.76(d,J=2.1Hz,2H),3.58(td,J=11.7,2.8Hz,1H),3.37–3.32(m,1H),2.97(t,J=6.3Hz,2H),2.05(h,J=5.9,5.4Hz,2H),1.98–1.89(m,2H),1.24(d,J=6.7Hz,3H).HRMS(ESI),m / z:389.2106[M+H+].

[0087] Example 8

[0088] A preparation method of an ATR inhibitor C05 compound. The synthesis method of compound C05 is the same as that of C01 in Example 4, and intermediate 6b obtained in Example 1 is used to replace intermediate 6a. It is a white solid with a yield of 86%. The NMR data of compound C05 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.20 (s, 1H), 8.12 (d, J = 7.5 Hz, 1H), 7.55–7.39 (m, 3H), 7.18 (t, J = 7.7 Hz, 1H), 5.44 (s, 1H), 5.13 (s, 1H), 4.30–4.11 (m, 4H), 4.03 (d, J = 10.8 Hz, 1H), 3.83 (d, J = 11.4 Hz, 1H), 3.75 (d, J = 10.3 Hz, 1H), 3.59 (t, J = 11.6 Hz, 1H), 3.47 (t, J = 12.8 Hz, 1H), 1.61 (s, 6H), 1.35 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 158.56, 152.36, 151.60, 151.30, 137.01, 130.02, 126.21, 125.77, 120.37, 120.30, 117.19, 112.96, 103.48, 73.73, 70.41, 66.47, 57.89, 41.48, 27.39, 27.34。

[0089] Example 9

[0090] A preparation method of an ATR inhibitor, compound C06. The synthesis method of compound C06 is the same as that of C01 in Example 4, except that intermediate 6b obtained in Example 1 is used to replace intermediate 6a, and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine is used to replace 4-indoleboronic acid pinacol ester. It is a white solid with a yield of 86%. The NMR data of compound C06 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.73 (s, 1H), 8.32 (d, J = 5.0 Hz, 1H), 8.02 (d, J = 5.1 Hz, 1H), 7.56 (s, 1H), 7.39 (s, 1H), 5.43 (s, 1H), 5.13 (s, 1H), 4.24 (t, J = 5.1 Hz, 2H), 4.16 (d, J = 4.9 Hz, 2H), 4.03 (d, J = 10.7 Hz, 1H), 3.83 (d, J = 11.5 Hz, 1H), 3.75 (d, J = 11.5 Hz, 1H), 3.59 (t, J = 11.6 Hz, 1H), 3.50 (d, J = 13.1 Hz, 1H), 1.61 (s, 6H), 1.36 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 156.64, 152.46, 152.07, 151.40, 150.16, 142.20, 137.23, 126.79, 126.62, 117.94, 117.68, 114.59, 102.12, 73.74, 70.36, 66.41, 57.87, 41.55, 27.35, 27.31。

[0091] Example 10

[0092] A preparation method of an ATR inhibitor C07 compound. The synthesis method of compound C07 is the same as that of C01 in Example 4, except that 4-indolylboronic acid pinacol ester is used instead of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine. A white solid is obtained by the reaction, and the yield is 84%. The NMR data of compound C06 are as follows: 11H NMR(400 MHz, DMSO-d6) δ 11.22 (s, 1H), 8.13 (d, J = 7.4 Hz, 1H), 7.49 (s, 2H), 7.44 (t, J = 2.7 Hz, 1H), 7.24–7.15 (m, 1H), 5.47 (s, 1H), 5.07 (s, 1H), 4.94 (s, 2H), 4.24 (d, J = 5.1 Hz, 2H), 4.17 (d, J = 5.2 Hz, 2H), 4.06–3.98 (m, 1H), 3.81 (d, J = 11.5 Hz, 1H), 3.73 (dd, J = 11.4, 3.1 Hz, 1H), 3.57 (td, J = 11.7, 11.2, 2.7 Hz, 1H), 3.47 (d, J = 13.2 Hz, 1H), 1.35 (d, J = 6.7 Hz, 3H). 13C NMR(101 MHz, DMSO-d6) δ 158.57, 152.27, 151.84, 144.77, 137.03, 129.94, 126.22, 125.79, 120.38, 120.35, 116.94, 113.03, 103.52, 70.38, 66.49, 64.46, 63.33, 41.08, 14.28. HRMS(ESI), m / z: 391.1890 [M+H+].

[0093] Example 11

[0094] A preparation method of an ATR inhibitor C08 compound. The synthesis method of compound C08 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole. A white solid is obtained by the reaction with a yield of 89%. The NMR data of compound C08 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 13.17 (s, 1H), 8.92 (s, 1H), 8.21 (d, J = 7.2 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.46 (t, J = 7.9 Hz, 1H), 5.56–5.34 (m, 1H), 5.07 (s, 1H), 4.95 (s, 2H), 4.28 (d, J = 5.1 Hz, 2H), 4.18 (d, J = 5.3 Hz, 2H), 4.02 (d, J = 10.9 Hz, 1H), 3.82 (d, J = 11.4 Hz, 1H), 3.73 (d, J = 11.4 Hz, 1H), 3.58 (t, J = 11.6 Hz, 1H), 3.49 (d, J = 13.3 Hz, 1H), 1.36 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 156.90, 152.35, 151.71, 145.18, 135.22, 131.28, 125.59, 121.05, 120.87, 117.42, 70.35, 66.45, 64.43, 63.30, 41.10, 14.30。

[0095] Example 12

[0096] A method for preparing an ATR inhibitor compound C09. The synthesis method of compound C09 is the same as that of C01 in Example 4, except that 7-azaindole-4-borate is used instead of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine. A white solid is obtained by the reaction with a yield of 83%. The NMR data of compound C09 are as follows: 1HNMR(400MHz, DMSO-d6) δ 11.73 (s, 1H), 8.32 (dd, J = 4.9, 2.1 Hz, 1H), 8.02 (dd, J = 4.8, 2.2 Hz, 1H), 7.56 (d, J = 2.9 Hz, 1H), 7.39 (d, J = 3.5 Hz, 1H), 5.46 (s, 1H), 5.24–5.00 (m, 1H), 4.95 (s, 2H), 4.26 (t, J = 5.3 Hz, 2H), 4.17 (d, J = 5.2 Hz, 2H), 3.81 (d, J = 11.5 Hz, 1H), 3.73 (d, J = 11.7 Hz, 1H), 3.58 (t, J = 12.1 Hz, 1H), 3.47 (t, J = 13.0 Hz, 1H), 1.36 (d, J = 6.6 Hz, 3H). 13C NMR(101MHz, DMSO-d6) δ 156.63, 152.35, 151.63, 145.54, 142.20, 137.17, 126.79, 117.69, 114.61, 102.16, 70.33, 66.43, 64.42, 63.26, 41.12, 14.31。

[0097] Example 13

[0098] A preparation method of an ATR inhibitor C10 compound. The synthesis method of compound C10 is the same as that of C01 in Example 4, except that 5-indolylboronic acid pinacol ester is used to replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine. A white solid is obtained by the reaction, and the yield is 76%. The NMR data of compound C10 are as follows: 1 H NMR(400MHz, DMSO-d6) δ 11.19 (s, 1H), 8.64 (s, 1H), 8.22 (d, J = 8.5 Hz, 1H), 7.44 (d, J = 8.7 Hz, 1H), 7.37 (d, J = 2.7 Hz, 1H), 6.55 (s, 1H), 5.47 (s, 1H), 5.07 (s, 1H), 4.92 (s, 2H), 4.24–4.18 (m, 2H), 4.15 (d, J = 5.1 Hz, 2H), 4.01 (dd, J = 11.3, 3.3 Hz, 1H), 3.80 (d, J = 11.5 Hz, 1H), 3.76–3.70 (m, 1H), 3.57 (td, J = 11.7, 2.6 Hz, 1H), 3.42 (t, J = 13.8 Hz, 1H), 1.34 (dd, J = 6.9, 2.0 Hz, 3H).

[0099] Example 14

[0100] A preparation method of an ATR inhibitor C11 compound. The synthesis method of compound C11 is the same as that of C01 in Example 4. Just replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with indole-6-boronic acid pinacol ester. A white solid is obtained by the reaction, and the yield is 82%. The NMR data of compound C11 are as follows: 1 HNMR(400MHz,DMSO-d6)δ11.24(s,1H),8.50(s,1H),8.13(d,J=8.4Hz,1H),7.58(d,J=8.4Hz,1H),7.43(d,J=2.9Hz,1H),6.46(s,1H),5.47(s,1H),5.07(s,1H),4.93(s,2H),4.21(d,J=5.2Hz,2H),4.16(d,J=5.1Hz,2H),4.02(d,J=11.1Hz,1H),3.81(d,J=11.5Hz,1H),3.77–3.70(m,1H),3.57(t,J=11.8Hz,1H),3.45(d,J=12.9Hz,1H),1.34(d,J=6.6Hz,3H。

[0101] Example 15

[0102] A preparation method of an ATR inhibitor C12 compound. The synthesis method of compound C12 is the same as that of C01 in Example 4. Just replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 7-indoleboronic acid pinacol ester. A white solid is obtained by the reaction, and the yield is 73%. The NMR data of compound C12 are as follows: 1 H NMR(400MHz,DMSO-d6)δ11.23(s,1H),8.25(d,J=7.5Hz,1H),7.69(d,J=7.8Hz,1H),7.48(d,J=2.9Hz,1H),7.14(t,J=7.6Hz,1H),6.56(s,1H),5.48(s,1H),5.07(s,1H),4.96(s,2H),4.42(q,J=4.4Hz,2H),4.20(t,J=5.2Hz,2H),4.02(dd,J=11.3,3.2Hz,1H),3.81(d,J=11.5Hz,1H),3.73(dd,J=11.6,3.2Hz,1H),3.58(td,J=11.6,2.6Hz,1H),3.46(t,J=13.4Hz,1H),1.35(d,J=6.7Hz,3H)。

[0103] Example 16

[0104] A preparation method of an ATR inhibitor C13 compound. The preparation method is as follows: Put the intermediate 6a (200 mg, 0.64 mmol) obtained in Example 1, 1-p-toluenesulfonylindole-3-boronic acid pinacol ester (307 mg, 0.77 mmol), Pd(Ph3P)4 (74 mg, 0.06 mmol) and K2CO3 (178 mg, 1.29 mmol) into a 25-ml three-necked flask, and add 8 ml of a mixed solvent of 1,4-dioxane / water (volume ratio 7 / 1). Under a nitrogen atmosphere, heat the above mixture to 100 °C and stir overnight. The next day, monitor by TLC plate that the raw materials have reacted completely. After cooling the reaction to room temperature, filter with diatomaceous earth to obtain the filtrate and concentrate to obtain a black viscous substance. Redissolve the crude product in 8 ml of methanol, then add 1 M NaOH aqueous solution, heat to 50 °C and stir for about 4 h. Concentrate under reduced pressure to remove the solvent, and finally purify by a rapid preparative column chromatograph to obtain 168 mg of a white compound C13, and the total yield of the two-step reaction is 84%. The NMR data of compound C13 are as follows: 1 H NMR(400MHz,DMSO-d6)δ11.49(s,1H),8.66–8.55(m,1H),8.13(d,J=2.8Hz,1H),7.51–7.40(m,1H),7.20–7.08(m,2H),5.45(s,1H),5.06(s,1H),4.91(s,2H),4.21(d,J=5.2Hz,2H),4.15(d,J=5.1Hz,2H),4.01(dd,J=11.4,3.3Hz,1H),3.80(d,J=11.4Hz,1H),3.72(dd,J=11.4,3.1Hz,1H),3.62–3.52(m,1H),3.48–3.38(m,1H),1.34(d,J=6.7Hz,3H).

[0105] Example 17

[0106] A preparation method of an ATR inhibitor C14 compound. The synthesis method of compound C14 is the same as that of C01 in Example 4, and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline is used instead of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine. A white solid is obtained by the reaction, and the yield is 80%. The NMR data of compound C14 are as follows: 11H NMR (400 MHz, CD3Cl) δ 7.39 (s, 2H), 7.19 (s, 5H), 6.70 (s, 4H), 5.03 (s, 4H), 4.74 (d, J = 14.9 Hz, 7H), 4.66 (s, 8H), 4.04 (s, 6H), 3.74–3.37 (m, 36H), 2.99 (s, 4H), 1.27 (s, 12H). 13 13C NMR (125 MHz, CD3Cl) δ 158.28, 156.93, 153.17, 148.21, 147.99, 130.66, 129.81, 129.67, 127.01, 115.13, 110.58, 72.43, 67.90, 66.67, 65.60, 48.55, 45.87, 43.95, 39.71, 31.61, 16.69。

[0107] Example 18

[0108] A preparation method of an ATR inhibitor C15 compound. The synthesis method of compound C15 is the same as that of C01 in Example 4. Just replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indol-2-one with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine. A white solid is obtained by the reaction with a yield of 78%. The NMR data of compound C15 are as follows: 1 1H NMR (400 MHz, CD3Cl) δ 10.87 (s, 1H), 8.56 (s, 1H), 7.65 (d, J = 32.0 Hz, 2H), 7.39 (s, 1H), 5.04 (s, 1H), 4.77 (d, J = 18.6 Hz, 2H), 4.66 (s, 2H), 4.05 (s, 2H), 3.64 (d, J = 16.7 Hz, 3H), 3.61–3.56 (m, 3H), 3.46 (d, J = 27.5 Hz, 2H), 1.17 (s, 3H). 13 13C NMR (100 MHz, CD3Cl) δ 175.92, 158.28, 156.93, 149.44, 148.21, 147.98, 130.19, 129.67, 128.60, 122.30, 121.62, 117.46, 72.42, 67.91, 66.66, 65.60, 48.55, 43.95, 42.48, 39.71, 16.69, -19.90。

[0109] Example 19

[0110] Preparation method of an ATR inhibitor C16 compound. The synthesis method of compound C16 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 5-fluoro-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole. A white solid is obtained by the reaction, and the yield is 88%. The NMR data of compound C16 are as follows: 1 H NMR(400MHz,DMSO-d6)δ11.23(s,1H),7.43(dt,J=9.2,3.4Hz,2H),7.03–6.95(m,1H),6.74(s,1H),5.36(s,1H),4.95(s,3H),4.15(d,J=4.1Hz,4H),3.96(dd,J=11.4,3.3Hz,1H),3.76(d,J=11.4Hz,1H),3.68(dd,J=11.5,3.1Hz,1H),3.58–3.49(m,1H),3.46–3.37(m,1H),1.32(d,J=6.7Hz,3H).

[0111] Example 20

[0112] Preparation method of an ATR inhibitor C17 compound. The synthesis method of compound C17 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 6-fluoro-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole. A white solid is obtained by the reaction, and the yield is 81%. The NMR data of compound C17 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.28 (s, 1H), 7.91 (dt, J = 11.6, 2.1 Hz, 1H), 7.50 (d, J = 3.0 Hz, 1H), 7.45 (t, J = 2.5 Hz, 1H), 7.29 (dd, J = 9.3, 2.5 Hz, 1H), 5.45 (s, 1H), 5.05 (s, 1H), 4.94 (s, 2H), 4.24 (d, J = 5.1 Hz, 2H), 4.16 (d, J = 5.1 Hz, 2H), 4.05–3.98 (m, 1H), 3.81 (d, J = 11.5 Hz, 1H), 3.73 (dd, J = 11.5, 3.2 Hz, 1H), 3.57 (t, J = 11.5 Hz, 1H), 3.46 (t, J = 13.5 Hz, 1H), 1.35 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 152.28, 151.69, 145.12, 137.01, 136.88, 130.95, 126.48, 123.07, 117.23, 108.02, 107.76, 103.66, 99.04, 98.79, 70.36, 66.46, 64.44, 63.29, 48.60, 41.09, 14.28。

[0113] Example 21

[0114] A method for preparing an ATR inhibitor C18 compound. The synthesis method of compound C18 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 6-trifluoromethyl-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole. A white solid is obtained as the product, and the yield is 60%. The NMR data of compound C18 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 11.68 (s, 1H), 8.38 (s, 1H), 7.83 (s, 1H), 7.71 (t, J = 2.5 Hz, 1H), 7.58 (d, J = 3.1 Hz, 1H), 5.46 (s, 1H), 4.96 (s, 3H), 4.28 (d, J = 5.2 Hz, 2H), 4.17 (d, J = 5.2 Hz, 2H), 4.04 (d, J = 10.8 Hz, 1H), 3.83 (d, J = 11.5 Hz, 1H), 3.74 (dd, J = 11.4, 3.1 Hz, 1H), 3.58 (t, J = 11.6 Hz, 1H), 3.48 (t, J = 12.9 Hz, 1H), 1.36 (d, J = 6.6 Hz, 3H).

[0115] Example 22

[0116] A preparation method of an ATR inhibitor C19 compound. The synthesis method of compound C19 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 6-methoxy-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole. A white solid is obtained by the reaction, and the yield is 55%. The NMR data of compound C19 are as follows: 1 H NMR(400MHz,DMSO-d6)δ10.99(s,1H),7.78(d,J=2.3Hz,1H),7.38(s,1H),7.28(s,1H),7.03(d,J=2.4Hz,1H),5.44(s,1H),5.04(s,1H),4.94(s,2H),4.25(t,J=5.3Hz,2H),4.17(d,J=5.1Hz,2H),4.09–3.98(m,1H),3.81(d,J=12.8Hz,4H),3.73(dd,J=11.6,3.1Hz,1H),3.64–3.51(m,1H),3.45(t,J=12.3Hz,1H),1.35(d,J=6.7Hz,3H).

[0117] Example 23

[0118] A preparation method of an ATR inhibitor C20 compound. The synthesis method of compound C20 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 6-(methoxycarbonyl)indole-4-boronic acid pinacol ester. A white solid is obtained by the reaction, and the yield is 86%. The NMR data of compound C20 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 8.75 (s, 1H), 8.15 (s, 1H), 7.70 (d, J = 2.9 Hz, 1H), 7.53 (d, J = 2.7 Hz, 1H), 5.47 (s, 1H), 5.07 (s, 1H), 4.95 (s, 2H), 4.26 (d, J = 5.3 Hz, 2H), 4.16 (t, J = 5.2 Hz, 2H), 4.07–3.97 (m, 1H), 3.90 (s, 3H), 3.82 (d, J = 11.5 Hz, 1H), 3.74 (dd, J = 11.5, 3.1 Hz, 1H), 3.59 (td, J = 11.6, 11.1, 2.5 Hz, 1H), 3.48 (t, J = 12.7 Hz, 1H), 1.37 (d, J = 6.7 Hz, 3H).

[0119] Example 24

[0120] A preparation method of an ATR inhibitor C21 compound, comprising the following steps: Dissolve the compound C20 (100 mg, 0.22 mmol) obtained in Example 23 in 10 ml of anhydrous THF under a N2 atmosphere, and add LiAlH4 (12 mg, 0.33 mmol) when cooled to 0 °C. Subsequently, seal the reaction system and heat it to 70 °C and stir overnight. The next day, monitor by TLC plate spotting, and the raw material has reacted completely. After cooling to room temperature, slowly add about 10 ml of water to quench the reaction. Extract the reaction solution with an ethyl acetate / water system, combine the organic phases collected three times, dry over anhydrous Na2SO4 solid, concentrate under reduced pressure and purify by flash column chromatography to obtain 80 mg of compound C21. White solid, with a yield of 80%. The NMR data of compound C21 is as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.14 (s, 1H), 8.09 (s, 1H), 7.47 (s, 1H), 7.43 (d, J = 2.4 Hz, 1H), 7.39 (t, J = 2.8 Hz, 1H), 5.49 (s, 1H), 5.15 (t, J = 5.7 Hz, 1H), 5.11–5.00 (m, 1H), 4.95 (s, 2H), 4.66 (d, J = 5.5 Hz, 2H), 4.25 (t, J = 5.1 Hz, 2H), 4.16 (t, J = 5.2 Hz, 2H), 4.03 (dd, J = 11.4, 3.4 Hz, 1H), 3.82 (d, J = 11.4 Hz, 1H), 3.73 (dd, J = 11.4, 3.1 Hz, 1H), 3.58 (td, J = 11.6, 2.6 Hz, 1H), 3.51–3.43 (m, 1H), 1.35 (d, J = 6.8 Hz, 3H). HRMS (ESI), m / z: 421.1992 [M+H+].

[0121] Example 25

[0122] A preparation method of an ATR inhibitor C22 compound. The synthesis method of compound C22 is the same as that of C01 in Example 4, except that 5-aminopyridine-3-boronic acid pinacol ester is used instead of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine. A pale yellow solid is obtained by the reaction, and the yield is 72%. The NMR data of compound C22 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 8.72 (d, J = 1.8 Hz, 1H), 7.98 (d, J = 2.7 Hz, 1H), 7.85 (t, J = 2.3 Hz, 1H), 5.44 (s, 3H), 5.02 (s, 1H), 4.93 (s, 2H), 4.16 (dq, J = 10.1, 6.0, 5.0 Hz, 4H), 3.99 (dd, J = 11.4, 3.3 Hz, 1H), 3.78 (d, J = 11.4 Hz, 1H), 3.70 (dd, J = 11.5, 3.1 Hz, 1H), 3.55 (td, J = 11.7, 2.7 Hz, 1H), 3.42 (d, J = 14.5 Hz, 1H), 1.32 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 155.40, 152.17, 151.68, 145.06, 144.47, 137.16, 136.98, 133.61, 118.55, 117.58, 70.33, 66.43, 64.40, 63.26, 41.12, 14.31.

[0123] Example 26

[0124] A preparation method of ATR inhibitor C23 compound. The synthesis method of compound C23 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 5-hydroxypyridine-3-boronic acid pinacol ester. A white solid was obtained by reaction with a yield of 76%. The NMR data of compound C23 are as follows: 1 H NMR(400MHz,DMSO-d6)δ10.13(s,1H),8.99(d,J=1.7Hz,1H),8.18(d,J=2.8Hz,1H),8.06(s,1H),5.42(s,1H),5.02(s,1H),4.94(s,2H),4.25–4.10(m,4H),4.00(dd,J=11.4,3.4Hz,1H),3.79(d,J=11.4Hz,1H),3.70(dd,J=11.5,3.2Hz,1H),3.56(dd,J=12.0,2.8Hz,1H),3.48–3.38(m,1H),1.33(d,J=6.7Hz,3H).

[0125] Example 27

[0126] A preparation method of ATR inhibitor C24 compound. The synthesis method of compound C24 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine with 2-aminopyrimidine-5-boronic acid pinacol ester to obtain a light gray solid with a yield of 74%. The NMR data of compound C24 are as follows: 1 HNMR(400MHz,DMSO-d6)δ9.06(s,2H),7.03(s,2H),5.40(s,1H),4.89(s,3H),4.22–4.06(m,4H),4.04–3.91(m,1H),3.76(d,J=11.5Hz,1H),3.73–3.63(m,1H),3.52(t,J=11.3Hz,1H),3.38(d,J=15.4Hz,1H),1.30(d,J=6.7Hz,3H).

[0127] Example 28

[0128] A preparation method of an ATR inhibitor C25 compound. The synthesis method of compound C25 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-ol with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-pyrrolo[2,3-c]pyridine to obtain a white solid with a yield of 72%. The NMR data of compound C25 are as follows: 1 H NMR(400MHz,DMSO-d6)δ9.35(s,1H),8.76–8.69(m,2H),5.44(s,1H),5.34(s,1H),5.03(s,1H),4.93(s,2H),4.21(d,J=5.2Hz,2H),4.15(d,J=5.0Hz,2H),4.00(dd,J=11.4,3.3Hz,1H),3.80(d,J=11.5Hz,1H),3.70(dd,J=11.5,3.2Hz,1H),3.60–3.50(m,1H),3.48–3.39(m,1H),1.52(s,6H),1.33(d,J=6.7Hz,3H).

[0129] Example 29

[0130] A preparation method of an ATR inhibitor C26 compound. The synthesis method of compound C26 is the same as that of C01 in Example 4. Replace 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-pyrrolo[2,3-c]pyridine with 3-(methylsulfonamido)phenylboronic acid pinacol ester to obtain a white solid with a yield of 79%. The NMR data of compound C26 are as follows: 1 HNMR(400MHz,DMSO-d6)δ9.85(s,1H),8.30(s,1H),8.11(d,J=7.7Hz,1H),7.43(t,J=7.9Hz,1H),7.32(dd,J=7.8,2.3Hz,1H),5.43(s,1H),5.05(s,1H),4.93(s,2H),4.18(d,J=4.6Hz,2H),4.15(d,J=4.5Hz,2H),4.00(dd,J=11.3,3.3Hz,1H),3.79(d,J=11.4Hz,1H),3.71(dd,J=11.5,3.1Hz,1H),3.56(td,J=11.7,2.7Hz,1H),3.42(t,J=11.2Hz,1H),3.02(s,3H),1.33(d,J=6.7Hz,3H).

[0131] Example 30

[0132] A preparation method of an ATR inhibitor C27 compound, comprising the following steps: Dissolve the intermediate 20a (100 mg, 0.26 mmol) obtained in Example 1 in 10 ml of methanol, add BrCN (42 mg, 0.39 mmol) at room temperature, and then stir overnight. The next day, monitor by TLC plate. The raw material has reacted completely. Concentrate under reduced pressure to remove the solvent methanol. Add 20 ml of a mixed solvent of ethyl acetate / water (volume ratio 1 / 1) thereto. Extract the reaction solution, combine the organic phases three times, dry over anhydrous Na2SO4, and then concentrate under reduced pressure again. Purify by flash column chromatography (dichloromethane / methanol = 97 / 3) to obtain 72 mg of compound C27. White solid, with a yield of 68%. The NMR data of compound C27 are as follows: 1 H NMR (400 MHz, DMSO-d6) δ 8.19 (d, J = 7.9 Hz, 1H), 7.72 (s, 2H), 7.29 (d, J = 7.8 Hz, 1H), 7.10 (td, J = 7.6, 1.2 Hz, 1H), 7.02 (td, J = 7.7, 1.3 Hz, 1H), 4.94 (s, 2H), 4.28–4.21 (m, 2H), 4.16 (d, J = 5.1 Hz, 2H), 4.02 (d, J = 8.9 Hz, 1H), 3.81 (d, J = 11.6 Hz, 1H), 3.72 (dd, J = 11.6, 3.1 Hz, 1H), 3.17 (d, J = 4.9 Hz, 4H), 1.37 (d, J = 6.8 Hz, 3H). HRMS (ESI), m / z: 407.1953 [M + H+].

[0133] Example 31

[0134] A preparation method of an ATR inhibitor C28 compound, comprising the following steps: Dissolve the intermediate 20a (100 mg, 0.26 mmol) obtained in Example 1 in 10 ml of 1,4-dioxane, add CDI (32 mg, 0.52 mmol) at room temperature, and then heat to 100 °C and stir for about 3 h. TLC plate monitoring shows that the raw material has reacted completely. Concentrate under reduced pressure to remove the solvent, and purify by flash column chromatography (dichloromethane / methanol = 48 / 2) to obtain 61 mg of compound C28. White solid, with a yield of 57%. The NMR data of compound C28 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 11.04 (s, 1H), 7.56 (d, J = 7.7 Hz, 1H), 7.10–6.96 (m, 3H), 4.94 (s, 4H), 4.15 (s, 4H), 4.01–3.92 (m, 1H), 3.76 (d, J = 11.5 Hz, 1H), 3.67 (dd, J = 11.5, 3.1 Hz, 1H), 3.52 (td, J = 11.7, 2.7 Hz, 1H), 3.40 (s, 1H), 1.32 (d, J = 6.8 Hz, 3H). HRMS (ESI), m / z: 408.1792 [M+H+].

[0135] Example 32

[0136] A method for preparing an ATR inhibitor C29 compound, comprising the following steps: Dissolve the intermediate 20a (100 mg, 0.26 mmol) obtained in Example 1 in 5 ml of pyridine, add methyl isothiocyanate (29 mg, 0.39 mmol) dropwise at room temperature, and then heat to 90 °C and stir for about 2 h. TLC monitoring shows that the raw materials have reacted completely to obtain intermediate 21b, which does not require further purification. After the reaction solution is cooled to room temperature, add EDCI (75 mg, 0.39 mmol) again, heat to 90 °C and stir overnight. The next day, TLC monitoring shows that intermediate 21b has reacted completely. Concentrate under reduced pressure and purify by flash column chromatography (dichloromethane / ethyl acetate = 3 / 2) to obtain 87 mg of compound C29. White solid, with a total yield of 79% for the two-step reaction. The NMR data of compound C29 are as follows: 11H NMR (400 MHz, DMSO-d6) δ 8.21 (dd, J = 10.2, 6.4 Hz, 2H), 7.26 (d, J = 7.8 Hz, 1H), 7.08 (td, J = 7.5, 1.2 Hz, 1H), 6.97 (td, J = 7.7, 1.3 Hz, 1H), 4.93 (s, 2H), 4.29–4.22 (m, 2H), 4.15 (t, J = 5.3 Hz, 2H), 4.05–3.98 (m, 1H), 3.81 (d, J = 11.5 Hz, 1H), 3.72 (dd, J = 11.6, 3.1 Hz, 1H), 3.63–3.45 (m, 2H), 3.34 (s, 2H), 3.08 (d, J = 4.8 Hz, 3H), 1.37 (d, J = 6.7 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 154.65, 152.64, 150.76, 150.64, 144.71, 142.87, 132.33, 122.41, 119.10, 115.61, 115.15, 113.72, 70.10, 66.19, 64.34, 63.16, 41.19, 29.44. HRMS (ESI), m / z: 421.2106 [M+H+].

[0137] Example 33

[0138] A preparation method of an ATR inhibitor C30 compound. The synthesis method of compound C30 is the same as the preparation method of compound C29 in Example 32, and only the intermediate 20b obtained in Example 1 needs to be used to replace the intermediate 20a. It is a white solid, and the total yield of the two-step reaction is 76%. The NMR data of compound C30 are as follows: 11H NMR (400 MHz, Chloroform-d) δ 8.51 (q, J = 5.0 Hz, 1H), 8.06 (dd, J = 7.5, 1.5 Hz, 1H), 6.95–6.85 (m, 2H), 5.28 (s, 2H), 4.95 (s, 2H), 4.22 (dt, J = 5.1, 2.9 Hz, 4H), 4.08 (dd, J = 11.4, 3.4 Hz, 1H), 3.85 (t, J = 2.5 Hz, 2H), 3.70 (td, J = 11.7, 11.2, 2.6 Hz, 1H), 3.64–3.54 (m, 1H), 3.25 (d, J = 4.9 Hz, 3H), 1.48 (d, J = 6.8 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 155.38, 153.24, 151.73, 151.03, 150.46, 144.08, 135.36, 135.28, 130.96, 119.59, 119.52, 116.54, 110.42, 110.38, 109.36, 109.19, 71.11, 67.15, 65.50, 63.94, 41.55, 29.80。

[0139] Example 34

[0140] A method for preparing an ATR inhibitor C31 compound. The synthesis method of compound C31 is the same as that of compound C29 in Example 32. A white solid can be obtained by using 20c to replace 20a. The total yield of the two-step reaction is 76%. The NMR data of compound C31 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 5.2 Hz, 1H), 8.15 (dd, J = 8.8, 5.3 Hz, 1H), 7.03 (dd, J = 9.7, 2.6 Hz, 1H), 6.79–6.71 (m, 1H), 4.93 (s, 4H), 4.27–4.20 (m, 2H), 4.16 (d, J = 5.0 Hz, 2H), 4.06–4.00 (m, 1H), 3.81 (d, J = 11.5 Hz, 1H), 3.72 (dd, J = 11.6, 3.1 Hz, 1H), 3.55 (dtd, J = 26.1, 14.7, 13.4, 7.3 Hz, 2H), 3.08 (d, J = 4.8 Hz, 3H), 1.36 (d, J = 6.8 Hz, 3H).

[0141] Example 35

[0142] A preparation method of an ATR inhibitor C32 compound. The synthesis method of compound C32 is the same as that of compound C29 in Example 32. A white solid can be obtained by replacing 20a with 20d. The total yield of the two-step reaction is 68%. The NMR data of compound C32 are as follows: 1 H NMR(400MHz,Chloroform-d)δ8.46–8.38(m,1H),8.05(dd,J=10.3,2.7Hz,1H),7.32(dd,J=8.6,5.0Hz,1H),6.88(ddd,J=9.5,8.6,2.7Hz,1H),5.25(s,2H),4.95(s,2H),4.21(ddt,J=8.2,5.3,2.6Hz,4H),4.09(dd,J=11.5,3.4Hz,1H),3.86(t,J=2.7Hz,2H),3.71(td,J=11.7,11.1,2.6Hz,1H),3.61(dt,J=13.8,6.8Hz,1H),3.21(d,J=5.0Hz,3H),1.49(d,J=6.8Hz,3H).13C NMR(101MHz,Chloroform-d)δ158.82,155.78,153.26,151.73,151.10,144.05,139.02,116.53,115.58,115.49,109.84,109.60,102.57,102.26,71.10,67.14,65.52,63.96,41.56,29.76。

[0143] Example 36

[0144] A preparation method of an ATR inhibitor C33 compound. The synthesis method of compound C33 is the same as that of compound C29 in Example 32. A white solid can be obtained by replacing 20a with 20e. The total yield of the two-step reaction is 64%. The NMR data of compound C33 are as follows: 11H NMR (400 MHz, Chloroform-d) δ 7.42 (q, J = 5.0 Hz, 1H), 7.25 (d, J = 7.6 Hz, 1H), 7.09 (td, J = 8.0, 4.7 Hz, 1H), 6.77 (dd, J = 11.4, 8.2 Hz, 1H), 5.39 (s, 2H), 4.95 (s, 2H), 4.18 (tt, J = 7.9, 3.2 Hz, 4H), 4.06–3.99 (m, 1H), 3.79 (t, J = 2.4 Hz, 2H), 3.63 (td, J = 11.8, 2.7 Hz, 1H), 3.56–3.45 (m, 1H), 3.16 (d, J = 5.0 Hz, 3H), 1.42 (d, J = 6.8 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 155.31, 153.12, 151.49, 150.18, 146.41, 146.37, 144.15, 123.27, 123.20, 117.06, 112.19, 112.16, 108.05, 107.84, 71.18, 67.34, 65.51, 63.97, 41.58, 29.72。

[0145] Example 37

[0146] A method for preparing an ATR inhibitor C34 compound. The synthesis method of compound C34 is the same as that of compound C29 in Example 32. A white solid can be obtained by using 20f instead of 20a. The total yield of the two-step reaction is 64%. The NMR data of compound C34 are as follows: 1 1H NMR (500 MHz, CDCl3) δ 7.59 (t, J = 1.0 Hz, 1H), 7.52 (d, J = 7.7 Hz, 1H), 7.18–7.09 (m, 1H), 5.93 (q, J = 4.4 Hz, 1H), 4.55–4.51 (m, 4H), 4.27 (ddd, J = 13.9, 6.2, 3.8 Hz, 1H), 4.06–3.99 (m, 1H), 3.93–3.81 (m, 6H), 3.71 (d, J = 6.2 Hz, 1H), 3.15 (d, J = 4.4 Hz, 3H), 1.30 (d, J = 8.3 Hz, 3H).

[0147] Example 38

[0148] A method for preparing an ATR inhibitor C35 compound. The synthesis method of compound C35 is the same as that of compound C29 in Example 32. A white solid can be obtained by using 20g instead of 20a. The total yield of the two-step reaction is 65%. The NMR data of compound C35 are as follows: 11H NMR (400 MHz, Chloroform-d) δ 8.55 (q, J = 5.1 Hz, 1H), 8.26 (d, J = 8.7 Hz, 1H), 7.30 (d, J = 1.2 Hz, 1H), 6.89 (ddd, J = 8.7, 2.4, 1.0 Hz, 1H), 5.07 (d, J = 57.0 Hz, 2H), 4.95 (s, 2H), 4.21 (dhept, J = 8.4, 3.0 Hz, 4H), 4.09 (dd, J = 11.4, 3.4 Hz, 1H), 3.90–3.81 (m, 2H), 3.70 (td, J = 11.7, 11.1, 2.6 Hz, 1H), 3.65–3.56 (m, 1H), 3.22 (d, J = 4.9 Hz, 3H), 1.48 (d, J = 6.8 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 156.55, 153.32, 151.75, 151.10, 145.40, 144.08, 143.93, 131.26, 116.58, 114.31, 112.41, 108.95, 71.10, 67.13, 65.52, 63.93, 41.56, 29.67。

[0149] Example 39

[0150] A method for preparing an ATR inhibitor C36 compound. The synthesis method of compound C36 is the same as that of compound C29 in Example 32. A white solid can be obtained by replacing 20h with 20a. The total yield of the two-step reaction is 71%. The NMR data of compound C36 are as follows: 11H NMR (400 MHz, Chloroform-d) δ 8.50 (q, J = 5.0 Hz, 1H), 8.17 (d, J = 8.8 Hz, 1H), 7.03 (d, J = 2.6 Hz, 1H), 6.63 (dd, J = 8.8, 2.6 Hz, 1H), 5.23 (s, 2H), 4.94 (s, 2H), 4.20 (dtt, J = 8.2, 5.2, 3.3 Hz, 4H), 4.08 (dd, J = 11.3, 3.4 Hz, 1H), 3.84 (d, J = 6.5 Hz, 5H), 3.70 (td, J = 11.7, 11.3, 2.6 Hz, 1H), 3.63–3.54 (m, 1H), 3.22 (d, J = 4.9 Hz, 3H), 1.47 (d, J = 6.8 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 156.56, 155.99, 153.31, 151.98, 151.20, 144.07, 143.79, 126.86, 116.29, 114.60, 106.78, 100.65, 71.13, 67.16, 65.52, 63.95, 55.80, 55.77, 41.50, 29.71。

[0151] Example 40

[0152] A method for preparing an ATR inhibitor C37 compound. The synthesis method of compound C37 is the same as that of compound C27 in Example 30. A white solid can be obtained by using 20b instead of 20a. The total yield of the two-step reaction is 65%. The NMR data of compound C37 are as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 8.18 (d, J = 8.1 Hz, 1H), 7.53 (s, 2H), 7.07 (t, J = 7.7 Hz, 1H), 6.98 (d, J = 8.2 Hz, 1H), 5.76 (s, 1H), 4.94 (s, 3H), 4.24 (s, 2H), 4.15 (s, 2H), 4.02 (d, J = 10.0 Hz, 1H), 3.82 (d, J = 11.6 Hz, 1H), 3.73 (d, J = 11.9 Hz, 1H), 3.57 (dd, J = 26.1, 14.8 Hz, 2H), 1.37 (d, J = 6.7 Hz, 3H).

[0153] Example 41

[0154] Detection of the enzyme activity of the compounds of the present invention and their in vitro anti-proliferation against tumor cells

[0155] Inhibition of ATM, ATR, DNA-PKcs and other kinase activities. Each kinase was separately reacted with the corresponding compounds C01 - C037 in a specified reaction solution. The reaction was initiated by adding a Mg(n) / ATP mixture and incubated at room temperature for a certain period of time. Then the reaction was terminated by adding phosphoric acid with a concentration of 0.5%. 10 μL of the terminated reaction solution was spotted onto a P30 filter pad and washed 4 times with 0.425% phosphoric acid solution for 4 minutes each time, and finally washed once with methanol, followed by drying and scintillation counting. The results are shown in Table 1. It can be seen from Table 1 that the compounds C01 - C037 of the present invention have a significant inhibitory effect on ATR.

[0156] Preferably, compounds C01, C02, C07, C09, C16, C17, C27, C29, C30, C31, C32, C37 in Table 1 with excellent ATR inhibition rates were used. Granta-519 cells were seeded in a 96-well plate at a density of 5×10 3 . The corresponding preferred ATR inhibitors (compounds C01 - C037) were added and co-incubated in an incubator with 5% carbon dioxide for 3 days. 10 μL of CCK8 solution was added and incubated for 1 h, and the absorbance at 450 nm was measured with an enzyme-linked immunosorbent assay (ELISA) reader. The ATR kinase inhibitor AZD-6738 was selected as the control group. The inhibition rate corresponding to the compound concentration was calculated according to the formula.

[0157] Inhibition rate = (absorbance value of the control group - absorbance value of the experimental group) / (absorbance value of the control group - absorbance value of the blank group) * 100%;

[0158] Finally, the half-maximal inhibitory concentration IC 50 was calculated by Graphpad prism 8.0 software. The results are shown in Table 2.

[0159] Table 1 Results of kinase activities

[0160]

[0161]

[0162]

[0163] In Table 1, * inhibition rate < 20%; ** inhibition rate 20% - 50%; *** inhibition rate 50% - 80%; **** inhibition rate > 80; among them, # indicates IC 50 200 - 500 nM; ## indicates IC 50 100 - 200 nM; indicates IC 50 50 - 100 nM: # indicates IC 50 < 50 nM.

[0164] Kinase Selectivity and Cellular Activity of Preferred Compounds in Table 2

[0165]

[0166]

[0167] In Table 2, #IC 50 > 2000 nM; ##IC 50 > 3000 nM; IC 50 > 5000 nM; *IC 50 500 - 1000 nM; **IC 50 200 - 500 nM; ***IC 50 100 - 200 nM; ****IC 50 50 - 100 nM: *****IC 50 < 50 nM.

[0168] Example 42

[0169] Kinase Selectivity Test

[0170] In a reaction tube, successively add buffer (8 mM MOPS, pH 7.0, 0.2 mM EDTA, 10 mM MnCl2), the kinase to be tested, the substrate of the kinase to be tested, 10 mM magnesium acetate and γ33P - ATP solution, as well as compounds of the present invention at different concentrations. Then add MgATP to the reaction to initiate the enzyme reaction process, and incubate at room temperature for 40 minutes. Finally, terminate the reaction with 5 μL of 3% phosphate buffer, titrate 10 μL of the reaction solution onto a Filtermat A membrane, wash it three times with 75 mM phosphate solution for 5 minutes each time, then wash it once with methanol, and finally dry the Filtermat A membrane and perform scintillation counting on it. Detailed experimental operations can be viewed on the website https: / / www.eurofinsdiscoveryservices.com. The numbers represent the remaining activity, and the smaller the number, the stronger the kinase activity.

[0171] At a concentration of 1 micromole per liter, a comprehensive selectivity evaluation of the compound was carried out on 226 human protein kinases related to oncology and their mutants. As Figure 1As shown, the compound C27 of the present invention has no significant inhibitory activity against most kinases, and only the inhibition rates of two kinases (cyclin-dependent kinase 9 (CDK9) and mammalian target of rapamycin (m-TOR)) exceed 50% (53% and 83% respectively). Overall, these results indicate that C27 has excellent kinase selectivity for ATR compared to other kinases widely related to oncology.

[0172] Example 43

[0173] In vitro anti-tumor activity test

[0174] LoVo and HCT116 cells were collected by centrifugation, then resuspended in complete DMED medium and counted. The cells were placed in a 24-well plate at a density of 400 cells per well. The 24-well plate was placed in an incubator containing 5% CO2 and cultured at 37 °C for three days. The preset concentration of the compound was co-incubated with LoVo and HCT116 cells for 1 hour, and the cells were placed in the incubator for one week. After one week, the cell supernatant was removed and the cells were washed twice with PBS, then methanol was slowly added and fixed at room temperature for 20 minutes. After removing the methanol, crystal violet was added to the dye and stained for 20 min, and then it was washed off with PBS. Subsequently, each well plate was photographed using a chemiluminescence imaging system, and the results of the clone experiment were analyzed using Image J for quantitative analysis, and finally used to calculate the corresponding inhibition rate and IC50 value.

[0175] As Figure 2 shown, the compound C27 has good anti-tumor activity in vitro, inhibits the proliferation of LoVo cells in a concentration-dependent manner, and additionally inhibits the proliferation of HCT116 cells in combination with the ATM inhibitor AZD1390.

[0176] References to "examples" in this document mean that the specific features, structures, or characteristics described in connection with the examples can be included in at least one embodiment of the present invention. The phrase appears in various places in the specification and does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment mutually exclusive with other embodiments. Those skilled in the art will explicitly and implicitly understand that the embodiments described herein can be combined with other embodiments.

[0177] Finally, it should be noted that: the embodiments disclosed in the present invention are only the preferred embodiments of the present invention, and are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An ATR inhibitor, characterized in that: Compounds having the following general structural formula I: The A and / or B is a nitrogen-containing heterocyclic group or heteroaryl group, or a nitrogen- and oxygen-containing heterocyclic group or heteroaryl group, or a nitrogen-, oxygen- and fluorine-containing heterocyclic group or heteroaryl group, or a nitrogen-, oxygen-, fluorine- and sulfur-containing heterocyclic group or heteroaryl group.

2. The ATR inhibitor according to claim 1, wherein: The A and B rings in General Formula I are R1 is any one or more of a hydrogen atom, a halogen, an alkyl group, and a substituted alkyl group.

3. The ATR inhibitor according to claim 1, wherein: Including the following structure:

4. A method for preparing an ATR inhibitor according to any one of claims 1-3, characterized in that: Including the following steps: (1) Using 2,4-dichloropurine as the starting material, reacting with dihydropyran (DHP) under acidic conditions to obtain intermediate 1, and then selectively reacting the chlorine at the 2-position of pyrimidine with (R)-3-methylmorpholine under the alkaline condition of K2CO3 to obtain a single-substituted intermediate 2; (2) Under anhydrous conditions, under N2 protection and at -78 °C, the H on the imidazole of intermediate 2 is removed by n-butyllithium, forming a relatively stable C anion with tetramethylethylenediamine in the solvent, and then adding DMF dropwise, finally forming aldehyde intermediate 3. Intermediate 3 is successively reduced by sodium borohydride and the tetrahydropyran (THP) is removed under acidic conditions to obtain intermediate 5; (3) 1,2-Dibromoethane is used as the ring-closing fragment group, and nucleophilic substitution reactions occur with the hydroxyl group and the amino group on both sides under alkaline conditions to obtain a tricyclic system intermediate 6 of morpholinoimidazopyrimidine; Intermediate 6 undergoes a Suzuki-Miyaura coupling reaction under the catalysis of Pd(Ph3P)4 to obtain the final product compound C01, compound C05-26; (4) The intermediate 6 is obtained as a nitro compound intermediate 19 through Buchwald-hartwig coupling reaction; Intermediate 19 is reduced to an amino-substituted benzene ring / heterocyclic derivative intermediate 20, and intermediate 20 directly reacts with cyanogen bromide (BrCN) to obtain compounds C27 and C37; Intermediate 20 reacts with N,N'-carbonyldiimidazole (CDI) to obtain compound C28; When intermediate 20 uses pyridine as the solvent and base, it reacts with methyl isothiocyanate to obtain a methylthiourea intermediate 21; The intermediate 21 uses 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) as a condensing agent and undergoes ring closure under heating conditions to obtain an N-methylbenzimidazole compound C29-36.

5. A method for preparing an ATR inhibitor according to any one of claims 1-3, characterized in that: (1) Using 2,6-dichloro-3-nitropyridin-4-amine as the starting material, first reacting with (R)-3-methylmorpholine in the presence of an organic base under heating to obtain a 2-position single-substituted intermediate 8 in a relatively high yield; (2) Subsequently, reducing the nitro group under the condition of Fe / HCl to obtain an aniline intermediate 9; Intermediate 9 and triethyl orthoformate undergo ring closure to obtain an imidazopyridine intermediate 10; Intermediate 10 reacts with DHP under acidic conditions to obtain intermediate 11; (3) Under anhydrous conditions, under N2 protection and at -78 °C, the H on the imidazole of intermediate 11 is removed by lithium diisopropylamide (LDA), forming a relatively stable C anion with tetramethylethylenediamine in the solvent, and then adding DMF dropwise, finally forming aldehyde intermediate 12. Intermediate 12 is successively reduced by sodium borohydride to obtain intermediate 13, intermediate 13 removes THP under acidic conditions to obtain intermediate 14, intermediate 14 undergoes ring closure under the conditions of potassium tert-butoxide and an aqueous solution of tetrabutylammonium bromide (TBAB) to obtain intermediate 15, and intermediate 15 undergoes a Suzuki-Miyaura coupling reaction under the catalysis of Pd(Ph3P)4 to obtain compound C02; ​ (4) The intermediate 11 is deprotonated in the presence of lithium diisopropylamide (LDA) and directly reacts with 1,3-dibromopropane or 1,4-dibromobutane to obtain the intermediate 16 with a single substituent, and the imidazopyridine nucleus with exposed NH is obtained after removing THP; cyclization is carried out under basic conditions to obtain the intermediate 18, and the intermediate 18 undergoes Suzuki-Miyaura coupling reaction under the catalysis of Pd(Ph3P)4 to obtain compounds C03 and C04.

6. The compound represented by the general formula I according to any one of claims 1-3, or its stereoisomer, or its pharmaceutically acceptable salt, for the treatment or improvement of gene defects and / or tumors.

7. The compound of general formula I as claimed in claim 1 for treating or ameliorating gene defects and / or tumors, or its stereoisomers, or its pharmaceutically acceptable salts as claimed in claim 6, characterized in that: The gene defect is ataxia telangiectasia mutated gene, and the tumors include gastric cancer solid tumors and colorectal cancer solid tumors, head and neck squamous cell carcinoma, small cell lung cancer with DDR mutation, ovarian cancer, leukemia, B-cell lymphoma, triple-negative breast cancer with HR mutation.

8. A pharmaceutical composition, characterized in that: It includes the compound represented by the general formula I according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt.

9. The pharmaceutical composition according to claim 8, wherein: It also includes pharmaceutically acceptable carriers, excipients, adjuvants and vehicles.

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