DNA-pk / parp1 dual-targeting inhibitors and preparation method and application thereof

Abstract

The application discloses a DNA-PK / PARP1 dual-target inhibitor and a preparation method and application thereof. The DNA-PK / PARP1 dual-target inhibitor is a compound with a structure as shown in the figure. The compound disclosed by the application is novel in structure, can selectively act on DNA-PK and PARP1, and exhibits good DNA-PK enzyme and PARP1 protein inhibition activity. The preparation method of the compound is mature, safe and pollution-free, and has the advantages of industrial promotion. The compound can be widely applied to preparation of a DNA-PK inhibitor and / or a PARP1 inhibitor, and preparation of a medicine for treating and / or preventing cancer.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmacy, and particularly relates to a DNA-PK / PARP1 dual-target inhibitor and a preparation method and application thereof. BACKGROUND

[0002] Cancer is a serious threat to human life and health, and is the second leading cause of human death. Effective treatment options for cancer have become the focus of scientists. In the past decade, a large number of molecular targeted drugs have been marketed for cancer treatment. However, since cancer is a multi-gene related and highly complex disease, single-target drugs often have limited efficacy in treating cancer and are only effective for some patients. In view of the above problems, a combination drug regimen is often used in clinical practice. Although the combination of drugs with different mechanisms of action can compensate for the shortcomings of single-target drugs to some extent, it also raises a series of problems, such as the superposition of toxic and side effects, and unpredictable PK / PD properties.

[0003] In recent years, significant progress has been made in the field of tumor treatment, one of the aspects that has attracted much attention is targeted therapy. DNA-PK (DNA-dependent protein kinase) is a protein kinase that plays an important role in DNA double-strand break repair. When DNA is damaged, DNA-PK is rapidly recruited to the damage site to initiate the repair mechanism. PARP1 (Poly(ADP-ribose) polymerase 1) is a poly-ADP-ribose polymerase that plays a key role in DNA damage response. When DNA is damaged, PARP1 is activated to catalyze the production of poly-ADP-ribose, which in turn recruits other repair proteins to participate in the DNA repair process.

[0004] The genome of tumor cells is usually unstable, and DNA damage occurs frequently. In order to survive, tumor cells will activate various DNA repair mechanisms, including DNA-PK and PARP1-mediated repair. If DNA-PK and PARP1 are inhibited simultaneously, the DNA repair ability of tumor cells will be further disrupted, leading to accumulation of DNA damage and ultimately inducing tumor cell death. DNA-PK / PAPR1 are both involved in the self-damage repair process of cancer cells, and simultaneous inhibition of DNA-PK / PARP1 has potential synergistic anticancer activity, especially in BRCA-mutated breast cancer cells.

[0005] DNA-PK / PARP1 dual-target inhibitors are a promising direction in the field of tumor treatment. By simultaneously inhibiting two key DNA repair proteins, tumor cells can be effectively killed, but the development of related drugs still needs to be improved. SUMMARY

[0006] In order to overcome the problems of the prior art, one of the purposes of the present application is to provide a compound, another purpose of the present application is to provide a preparation method of the compound, a third purpose of the present application is to provide a pharmaceutical composition, and a fourth purpose of the present application is to provide an application of the compound. In order to achieve the above purposes, the technical scheme adopted by the present application is:

[0007] The first aspect of the present application provides a compound, the structure of the compound is shown in formula 1:

[0008]

[0009] In formula 1, R1 is selected from any one of alkyl, aryl, heterocycle, and R2 is selected from any one of alkyl, aryl, heterocycle.

[0010] Preferably, in formula 1, R1 is selected from any one of alkyl and aryl, and R2 is selected from any one of alkyl and heterocycle.

[0011] Preferably, the compound includes any one of the following compounds (1) to (10):

[0012]

[0013] Preferably, the compound is a DNA-PK and PARP1 dual-target inhibitor.

[0014] The second aspect of the present application provides a preparation method of the compound according to the first aspect of the present application, including the following steps:

[0015] 7) The compound shown in formula I is mixed with 1-boc piperazine, the protecting group is removed, and then reacted with an amino acid to obtain a compound shown in formula II;

[0016]

[0017] In the formula, R2 is selected from any one of alkyl, aryl and heterocycle;

[0018] 8) The compound shown in formula III is mixed with bromo acid methyl ester, and then reduced to obtain a compound shown in formula IV;

[0019]

[0020] In the formula, R1 is selected from any one of alkyl, aryl and heterocycle;

[0021] 9) The compound shown in formula V is reacted with 4-amino tetrahydro pyran hydrochloride, and then hydrolyzed to obtain a compound shown in formula VI;

[0022]

[0023] 10) reacting the compound of formula VI with diphenyl phosphorazide, and then with iodomethane to obtain the compound of formula VII;

[0024]

[0025] 11) reacting the compound of formula VII with the compound of formula IV, and then hydrolyzing to obtain the compound of formula VIII:

[0026]

[0027] 12) reacting the compound of formula II with the compound of formula VIII to obtain the target compound.

[0028] Preferably, in the step 1), the molar ratio of the compound of formula I to 1-boc piperazine is 1:0.8-1.2, more preferably 1:1.

[0029] Preferably, in the step 2), the molar ratio of the compound of formula III to bromoformic acid methyl ester is 1:1.1-1.5, more preferably 1:1.39.

[0030] Preferably, in the step 3), the molar ratio of the compound of formula V to 4-aminotetrahydropyran hydrochloride is 1:0.8-1.2, more preferably 1:1.

[0031] Preferably, in the step 4), the molar ratio of the compound of formula VI to diphenyl phosphorazide is 1:0.8-1.2, more preferably 1:1.

[0032] Preferably, in the step 5), the molar ratio of the compound of formula VII to the compound of formula IV is 1:1-1.4, more preferably 1:1.2.

[0033] Preferably, in the step 6), the molar ratio of the compound of formula II to the compound of formula VIII is 1:1-1.4, more preferably 1:1.2.

[0034] The third aspect of the present application provides a pharmaceutical composition comprising the compound of the first aspect of the present application or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt or co-crystal thereof.

[0035] The fourth aspect of the present application provides use of the compound of the first aspect of the present application or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt or co-crystal thereof in the preparation of a medicament for treating and / or preventing and / or delaying and / or assisting the treatment of cancer.

[0036] Preferably, the cancer comprises at least one of breast cancer, melanoma, leukemia.

[0037] Preferably, the cancer drug comprises a DNA-PK inhibitor and / or a PARP1 inhibitor.

[0038] The present application has the following advantages and effects over the prior art:

[0039] The compound disclosed by the present application has a novel structure, can selectively act on DNA-PK and PARP1, and exhibits good DNA-PK enzyme and PARP1 protein inhibition activity; the preparation method of the compound is mature, safe and pollution-free, and has the advantages of industrial promotion; the compound can be widely used for preparing a DNA-PK inhibitor and / or a PARP1 inhibitor, and a drug for treating and / or preventing cancer. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 A chemical structure diagram of the compound of the present application is shown in the following figure:

[0041] Figure 2 A preparation reaction route diagram of the compound of the present application is shown in the following figure. DETAILED DESCRIPTION

[0042] The specific implementation of the present application is further described below in combination with examples, but the implementation and protection of the present application are not limited thereto. It should be noted that if the following processes are not specifically described in detail, they can be implemented or understood by referring to the prior art by those skilled in the art. If the reagents or instruments used are not specified by the manufacturer, they are considered to be conventional products that can be obtained by market purchase.

[0043] The chemical structure of the compound of the present application is shown in the following figure: Figure 1

[0044] The preparation reaction route of the compound of the present application is shown in the following figure: Figure 2 According to the above reaction route, the preparation of the compound comprises the following steps:

[0045] Route A:

[0046]

[0047] According to the above reaction route, the preparation of the compound comprises the following steps:

[0048] 1. 4-methyl-3-nitrophenol (A) is dissolved in an N,N-dimethylformamide solution, 2.5 molar times of potassium carbonate is added according to the molar amount of 4-methyl-3-nitrophenol, and the reaction is carried out at 80°C for 30 minutes. Then 1.39 molar times of compound B is added according to the molar amount of 4-methyl-3-nitrophenol, and the reaction is carried out at 80°C for 30 minutes. After monitoring the completion of the reaction on a thin layer chromatography plate, the reaction liquid is extracted with ethyl acetate, and the organic phase is dried with saturated brine and anhydrous sodium sulfate, and then vacuum desolventized to obtain an intermediate C. The specific reaction formula is as follows:​

[0049]

[0050] 2. Take the intermediate C obtained in step 1 and dissolve it in methanol, add 2 scoops of Raney nickel under ice bath conditions, and add 2.5 molar times of hydrazine hydrate based on the molar amount of the intermediate. React at room temperature for 5 minutes, monitor the reaction completion with a thin layer chromatography plate, desolventize, and purify by column chromatography to obtain intermediate D. The specific reaction formula is as follows:

[0051]

[0052] Route B:

[0053]

[0054] According to the above reaction route, the preparation of the compound comprises the following steps;

[0055] 1. Dissolve ethyl 2,4-dichloro-5-pyrimidinecarboxylate (E) in acetonitrile, add 1 molar times of 4-aminotetrahydropyran hydrochloride (F) based on the molar amount of ethyl 2,4-dichloro-5-pyrimidinecarboxylate, and add 2.5 molar times of potassium carbonate based on the molar amount of ethyl 2,4-dichloro-5-pyrimidinecarboxylate, and react at room temperature for 10 hours. Monitor the reaction completion with a thin layer chromatography plate. Desolventize, purify by column chromatography, and obtain intermediate G. The specific reaction formula is as follows:

[0056]

[0057] 2. Take the intermediate G obtained in step 1 and dissolve it in tetrahydrofuran, add 2.5 equivalents of sodium hydroxide solution, and react at 60°C oil bath for 1 hour. Monitor the reaction completion with a thin layer chromatography plate, and stop the reaction. Distill off the solvent under reduced pressure to obtain a crude product. Add water to the crude product, adjust the pH to weakly acidic (PH≈5-6) with dilute hydrochloric acid under ice bath conditions. Precipitate, and filter out the white solid, which is intermediate H. The specific reaction formula is as follows:

[0058]

[0059] 3. Take the intermediate H obtained in step 2 and dissolve it in DMA, add 1 molar times of diphenyl phosphorazide and 1 molar times of triethylamine, and react at room temperature for one hour. React at 120°C oil bath for 12 hours. Monitor the reaction completion with a thin layer chromatography plate, stop the reaction, quickly stir with ice water, precipitate, and filter out the light yellow solid, which is intermediate I. The specific reaction formula is as follows:

[0060]

[0061] 4. The intermediate I obtained in step 3 is dissolved in anhydrous tetrahydrofuran, and 2 moles of sodium hydride is added under ice bath condition. After the ice is removed, the reaction is allowed to proceed at room temperature for 30 minutes. Iodomethane is added, and the reaction is allowed to proceed at room temperature for 8 hours. After the reaction is completed as monitored by thin layer chromatography, the reaction is stopped. The reaction solution is extracted with ethyl acetate, and the organic phase is dried with saturated brine and anhydrous sodium sulfate, and then distilled under reduced pressure. The intermediate J is obtained by column chromatography. The specific reaction formula is as follows:

[0062]

[0063] Scheme C:

[0064]

[0065] 1. The amino acid (M) is dissolved in tetrahydrofuran, and 2 moles of sodium hydroxide is added. Water (THF: water = 1:1) and di-carbonic acid anhydride are added under ice bath condition. After the ice is removed, the reaction is allowed to proceed at room temperature for 8 hours. After the reaction is completed as monitored by thin layer chromatography, the reaction is stopped. The reaction solution is extracted with petroleum ether, and the water phase is adjusted to soft acidity (pH ≈ 5-6) with dilute hydrochloric acid under ice bath condition. The reaction is extracted with ethyl acetate, and the organic phase is dried with saturated brine and anhydrous sodium sulfate, and then distilled under reduced pressure to obtain the intermediate N. The specific reaction formula is as follows:

[0066]

[0067] 2. The 2-fluoro-5-((4-oxo-3,4-dihydrophthalidin-1-yl)methyl)benzoic acid (O) is dissolved in DMF, and 1 mole of piperazine-1-carboxylic acid tert-butyl ester acetate (P) is added according to the molar amount of the compound O. 1.5 moles of HATU and 2.5 moles of N,N-diisopropyl ethylamine are added, and the reaction is allowed to proceed at room temperature for 3 hours. After the reaction is completed as monitored by thin layer chromatography, the reaction is stopped. Ice water is added, and the slurry is filtered under reduced pressure to obtain the pink intermediate Q. The specific reaction formula is as follows:

[0068]

[0069] 3. The intermediate Q obtained in step 2 is dissolved in dichloromethane, and an appropriate amount of HCl·1,4 dioxane is added at room temperature. After the reaction is completed as monitored by thin layer chromatography, the precipitated solid is filtered under reduced pressure to obtain the pink intermediate R. The specific reaction formula is as follows:

[0070]

[0071] 4. Take the intermediate N obtained in step 1, and the intermediate R obtained in step 3, add 1.5 moles of intermediate R per mole of intermediate N, add 1.5 moles of HATU per mole of intermediate N, and add 2.5 moles of N,N-diisopropyl ethylamine per mole of intermediate N, and react at room temperature for 3 hours. After the reaction is completed, as monitored by TLC, stop the reaction, extract the reaction solution with ethyl acetate, dry the organic phase with saturated brine and anhydrous sodium sulfate, and distill under reduced pressure to obtain white intermediate S. The specific reaction formula is as follows:

[0072]

[0073] 5. Dissolve the intermediate S obtained in step 4 in dichloromethane, add an appropriate amount of HCl-1,4-dioxane at room temperature, and monitor the reaction completion by TLC. After the reaction is completed, the solid is precipitated and dried under reduced pressure to obtain intermediate T. The specific reaction formula is as follows:

[0074]

[0075] Scheme D:

[0076]

[0077] 1. Dissolve the intermediate J in a 1,4-dioxane solution, add 1.2 moles of intermediate D per mole of intermediate J, add 0.1 moles of tris(dibenzylideneacetone)dipalladium per mole of intermediate J, add 0.3 moles of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene per mole of intermediate J, and add 2.5 moles of cesium carbonate per mole of intermediate J, and react at 105°C for 12 hours under a nitrogen atmosphere. After the reaction is completed, as monitored by TLC, extract the reaction solution with dichloromethane, dry the organic phase with saturated brine and anhydrous sodium sulfate, and separate and purify by column chromatography to obtain intermediate K. The specific reaction formula is as follows:

[0078]

[0079] 2. Dissolve the intermediate K in a mixture of methanol and water, add 2.5 moles of sodium hydroxide per mole of intermediate K, and react at room temperature for 30 minutes. After the reaction is completed, as monitored by TLC, stop the reaction. Distill the solvent under reduced pressure to obtain a crude product. Add water to the crude product, and adjust the pH to weakly acidic (pH ≈ 5-6) using dilute hydrochloric acid under ice bath conditions. Filter out the white solid, which is intermediate L. The specific reaction formula is as follows:

[0080]

[0081] 3. Take the intermediate L obtained in step 2, add an appropriate amount of DMF as solvent, then add 1.2 molar times of intermediate T, 1.2 molar times of EDCI, 1.5 molar times of HOBT and 2.5 molar times of triethylamine according to the molar amount of intermediate L. Under normal temperature conditions, react for 2 hours, monitor the reaction completion by TLC plate, then add ice water, beat and filter to obtain the final product. The specific reaction formula is as follows:

[0082]

[0083] Example 1

[0084] The specific preparation steps of the compound of the present example are as follows:

[0085] 1) Synthesis of intermediate 5-(4-methyl-3-nitrophenoxy)pentanoic acid methyl ester (C): 1.0 g of 4-methyl-3-nitrophenol (A) (6.53 mmol) was dissolved in 10 ml of N,N-dimethylformamide solution, 2.26 g of potassium carbonate (16.33 mmol) was added, and the reaction was carried out at 80°C for 30 minutes, then 1.64 g of 5-bromomethyl pentanoate (B) (9.08 mmol) was added, and the reaction was carried out for 30 minutes. After monitoring the completion of the reaction by TLC plate, the reaction solution was extracted with ethyl acetate (30 mL x 3), and the organic phase was separated and washed with water (5 mL x 1), saturated brine (5 mL x 3), then dried with anhydrous sodium sulfate, filtered, and the ethyl acetate was removed under reduced pressure to obtain a solid 1.55 g, with a yield of 93.9%. The solid was identified by nuclear magnetic resonance method, and the results showed that the solid was 5-(4-methyl-3-nitrophenoxy)pentanoic acid methyl ester (C). Its structural formula is:

[0086]

[0087] 2) Synthesis of intermediate 5-(4-methyl-3-nitrophenoxy)pentanoic acid methyl ester (C): 1.55 g of 5-(4-methyl-3-nitrophenoxy)pentanoic acid methyl ester (C) was taken in a reaction bottle, dissolved in 15 mL of methanol, and 2 scoops of Raney nickel and 957.47 mg of 80% Raney nickel (0.147 mmol) were added in turn under ice bath conditions. The reaction was carried out at room temperature for 30 minutes, and after monitoring the completion of the reaction by TLC plate, the methanol was removed from the reaction solution under reduced pressure to obtain a solid, which was column chromatographed with dichloromethane:methanol to obtain a white solid 1.2 g, with a yield of 87.6%. The white solid obtained by column chromatography was identified by nuclear magnetic resonance method, and the results showed that the white solid was 5-(4-methyl-3-nitrophenoxy)pentanoic acid methyl ester (C). Its structural formula is:

[0088]

[0089] 3) Synthesis of 2-chloro-4-((tetrahydro-2h-pyran-4-yl)amino)pyrimidine-5- carboxylic acid ethyl ester (G): Take 5 g (22.62 mol) of 2,4-dichloro-5- pyrimidine carboxylic acid ethyl ester (E) in a reaction bottle, add 3.11 g (22.62 mol) of 4-aminotetrahydropyran hydrochloride (F), add 7.82 g (56.55 mol) of potassium carbonate, and dissolve in 30 ml of acetonitrile. React at room temperature for 10 hours, and after monitoring the completion of the reaction using a thin layer chromatography plate, wash the insoluble solid with 25 ml each of dichloromethane and ethanol, remove the liquid, and obtain a solid. Column chromatograph the solid using dichloromethane: petroleum ether to obtain 6 g of a white solid with a yield of 87.6%. Identify the white solid obtained by column chromatography using nuclear magnetic resonance, and the results show that the white solid is 5-(4-methyl-3-aminophenoxy)pentanoic acid methyl ester (G). The structural formula is:

[0090]

[0091] 4) Synthesis of 2-chloro-4-((tetrahydro-2h-pyran-4-yl)amino)pyrimidine-5- carboxylic acid (H): Dissolve 6 g (21 mol) of 2-chloro-4-((tetrahydro-2h-pyran-4- yl)amino)pyrimidine-5-carboxylic acid ethyl ester (G) in tetrahydrofuran, add 1.68 g (41.07 mol) of sodium hydroxide dissolved in 5 ml of water, and then add to the reaction solution. Place the reaction bottle in a 60°C oil bath and react for 1 hour. After monitoring the completion of the reaction using a thin layer chromatography plate, stop the reaction. Distill off the solvent from the reaction solution under reduced pressure to obtain a crude product. Add water to the crude product, and then adjust the pH to weak acidity (pH ≈ 5-6) using dilute hydrochloric acid under ice bath conditions. Filter out the white solid precipitate to obtain 4.4 g of a white solid with a yield of 81%. Identify the solid obtained using nuclear magnetic resonance, and the results show that the solid is 2-chloro-4-((tetrahydro-2h-pyran-4-yl)amino)pyrimidine-5-carboxylic acid (H). The structural formula is:

[0092]

[0093] 5) Synthesis of 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (I): Take 2 g (7.76 mol) of 2-chloro-4-((tetrahydro-2h-pyran-4-yl)amino)pyrimidine-5-carboxylic acid (H) dissolved in 2 ml of DMA, add 1.67 ml (7.76 mol) of diphenyl phosphorazide and 1.08 ml (7.76 mol) of triethylamine, and place in an oil bath at 120°C for 12 hours. After monitoring the reaction completion on a thin layer chromatography plate, stop the reaction, quickly stir with ice water, and precipitate the solid. Filter out the light yellow solid 0.92 g, and the yield is 46.54%. Identify the obtained solid by nuclear magnetic resonance method, and the result shows that the solid is 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (I). Its structural formula is:

[0094]

[0095] 6) Synthesis of 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (J): Take 920 mg (3.61 mol) of 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (I) dissolved in anhydrous tetrahydrofuran, add 362 mg (9.03 mol) of sodium hydride under ice bath condition, remove the ice after the reaction, and place in an oil bath at room temperature for 30 min. Then, add 563 μl (9.03 mol) of methyl iodide, and continue the reaction at room temperature for 8 hours. After monitoring the reaction completion on a thin layer chromatography plate, stop the reaction. Extract the reaction liquid with ethyl acetate, dry the organic phase with saturated brine and anhydrous sodium sulfate, desolventize, and separate and purify by column chromatography to obtain 673 mg of white solid, and the yield is 69.33%. Identify the obtained solid by nuclear magnetic resonance method, and the result shows that the solid is 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (J). Its structural formula is:

[0096]

[0097] 7) Synthesis of 4-((tert-butoxycarbonyl)amino)butyric acid (N): 500 mg (4.85 mol) of 4-aminobutyric acid (M) was dissolved in 5 ml of tetrahydrofuran, 484 mg (12.12 mol) of sodium hydroxide was added, 5 ml of water was added under ice bath, 1.17 ml (5.09 mol) of acetic anhydride was slowly added dropwise, and the reaction was allowed to proceed for 8 hours. After monitoring the reaction completion by thin layer chromatography, the reaction solution was extracted with ethyl acetate (30 ml x 1), and the aqueous phase was adjusted to be acidic, and then extracted with ethyl acetate (30 ml x 3). The organic phase was washed with water (5 ml x 1), saturated brine (5 ml x 3), and then dried with anhydrous sodium sulfate, filtered, and then the ethyl acetate was removed under reduced pressure to obtain 830 mg of a white solid, and the yield was 84.23%. The white solid was identified by nuclear magnetic resonance method, and the results showed that the white solid was 4-((tert-butoxycarbonyl)amino)butyric acid (N). The structural formula is:

[0098]

[0099] 8) Synthesis of tert-butyl 4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazine-1-yl)methyl)benzoyl)piperazine-1-carboxylate (Q): 2 g (6.71 mol) of 2-fluoro-5-((4-oxo-3,4-dihydrophthalazine-1-yl)methyl)benzoic acid (O) was dissolved in DMF, 1.25 g (6.71 mol) of tert-butyl piperazine-1-carboxylate acetate (P) was added, 3.82 g (10.06 mol) of HATU was added, 2.34 ml (13.41 mol) of N,N-diisopropyl ethylamine was added, and the reaction was allowed to proceed at room temperature for 3 hours. After monitoring the reaction completion by thin layer chromatography, the reaction was stopped, ice water was added, and the slurry was filtered under reduced pressure to obtain 2.7 g of a pink solid, and the yield was 86.31%. The solid was identified by nuclear magnetic resonance method, and the results showed that the solid was tert-butyl 4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazine-1-yl)methyl)benzoyl)piperazine-1-carboxylate (Q). The structural formula is:

[0100]

[0101] 9) Synthesis of 4-(4-fluoro-3-(piperazin-1-carbonyl)benzyl)phthalazin-1(2H)-one (R): Take 2.7 g of tert-butyl 4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1- yl)methyl)benzoyl)piperazine-1-carboxylate (Q) dissolved in 6 ml of dichloromethane, add 3 ml of HCl-1,4 dioxane at room temperature, monitor the reaction completion by TLC plate, after the solid precipitates, reduce pressure and filter to obtain a pink solid 1.6 g, yield 75.45%. The solid obtained was identified by nuclear magnetic resonance method, the results showed that the solid was 4-(4-fluoro-3-(piperazin-1-carbonyl)benzyl)phthalazin-1(2H)-one (R). Its structural formula is:

[0102]

[0103] 10) Synthesis of tert-butyl (4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1- yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)carbamate (S): Take 830 mg (4.08 mol) of 4-((tert-butoxycarbonyl)amino)butanoic acid (N), add 1.5 g (4.08 mol) of 4-(4-fluoro-3-(piperazin-1-carbonyl)benzyl)phthalazin-1(2H)-one (R), add 2.33 g (6.13 mol) of HATU, 1.78 (10.21 mol) of N,N-diisopropylethylamine, react at room temperature for 3 hours, stop the reaction after monitoring the reaction completion by TLC plate, extract the reaction liquid with ethyl acetate, dry the organic phase with saturated brine and anhydrous sodium sulfate, and distill under reduced pressure to obtain a white intermediate 1.85 g, yield 82.12%. The solid obtained was identified by nuclear magnetic resonance method, the results showed that the solid was tert-butyl (4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1- yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)carbamate (S). Its structural formula is:

[0104]

[0105] 11) Synthesis of 4-(3-(4-(4-aminobutanoyl)piperazine-l-carbonyl)-4- fluorobenzyl)phthalazine-l(2H)-one (T): Take 1.85 g (3.35 mol) of tert-butyl (4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazine-l-yl)methyl)benzoyl)piperazin- 1-yl)-4-oxobutyl)carbamate (S) dissolved in 4 ml of dichloromethane, add 4 ml of HCl-1,4 dioxane at room temperature, monitor the reaction completion by TLC plate, after the solid precipitates, reduce pressure and filter at low temperature to obtain 1.1 g of pink solid, with a yield of 72.64%. The solid obtained is identified by nuclear magnetic resonance method, the results show that the solid is 4-(3-(4-(4-aminobutanoyl)piperazine-l-carbonyl)-4- fluorobenzyl)phthalazine-l(2H)-one (T), and its structural formula is:

[0106]

[0107] 12) Synthesis of intermediate methyl 5-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro- 2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanoate (K): Take 0.4 g of 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8- one (J) (1.49 mmol) in a reaction bottle, add 0.365 g of methyl 5-(4-methyl-3- aminophenoxy)pentanoate (1.64 mmol), 0.258 g of 4,5-bis(diphenylphosphino)-9,9- dimethylxanthene (0.447 mmol), 0.97 g of cesium carbonate (2.98 mmol), 0.12 g of tris(dibenzylideneacetone)dipalladium, and 15 mL of 1,4-dioxane solution, and react at 105°C for 12 hours under a nitrogen atmosphere. After monitoring the reaction completion by TLC plate, the reaction liquid is extracted with ethyl acetate (30 mL x 3), and then separated by standing. The organic phase is washed with water (5 mL x 2), saturated brine (5 mL x 5) in sequence, and then dried with anhydrous sodium sulfate, filtered, and then the ethyl acetate is removed under reduced pressure to obtain a solid. The solid is subjected to column chromatography with dichloromethane:methanol to obtain 0.3 g of white solid, with a yield of 44.2%. The colorless liquid obtained by column chromatography is identified by nuclear magnetic resonance method, and the results show that the white solid is methyl 4,4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9- dihydro-7H-purin-2-yl)amino)phenoxy)pentanoate (K). Its structural formula is:

[0108]

[0109] 13) Synthesis of intermediate 5-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)- 8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanoic acid (L): Take 0.3 g of 4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2- yl)amino)phenoxy)pentanoic acid methyl ester (K) in a reaction bottle (0.658 mmol), dissolve with 5 ml of methanol. Add 0.065 g of NaOH (1.65 mmol), dissolve with 2 ml of water. Slowly drop the aqueous NaOH solution into the reaction bottle. React for 30 minutes under the condition of 60 degrees. After monitoring the reaction completion by thin layer chromatography, remove the solvent from the reaction liquid by reduced pressure distillation to obtain a crude product. After adding water to the crude product, stir quickly, and extract by suction filtration to obtain 260 mg of white solid, with a yield of 89.7%. Identify the white solid obtained by suction filtration by nuclear magnetic resonance method, and the result shows that the white solid is 4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanoic acid (L). Its structural formula:

[0110]

[0111] 14) Synthesis of product N-(4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophtalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)-5-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanamide: Take 0.04 g of intermediate 5-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanoic acid (L) (87.81 μmol) in a reaction bottle, add 0.039 g of 4-(3-(4-(4-aminobutanoyl)piperazine-1-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one (T) (87.81 μmol), 0.012 g of EDCI (105.38 μmol), 0.018 g of HOBT (131.72 μmol), 0.022 g of triethylamine (219.53 μmol) and 1 ml of DMF in turn. Reaction at room temperature for 2 hours, TLC plate monitoring reaction complete, add ice water to the reaction bottle, stir quickly, suction filtration to obtain 0.015 g of white solid, yield 19.2%. The white solid obtained by suction filtration was identified by nuclear magnetic resonance method, and the results showed that the white solid was N-(4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophtalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)-5-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanamide. Its structural formula:

[0112]

[0113] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum and mass spectrum of compound LSQ-1 are as follows: 1H NMR (400 MHz, CDC13) δ 8.44 - 8.32 (m, 1H), 7.84 (d, J = 1.7 Hz, 1H), 7.75 (d, J = 2.4 Hz, 1H), 7.69 (dt, J = 5.4, 2.0 Hz, 2H), 7.64 (dt, J = 8.4, 3.7 Hz, 1H), 7.34 - 7.21 (m, 2H), 7.02 - 6.86 (m, 2H), 6.81 (s, 1H), 6.43 (dd, J = 8.3, 2.8 Hz, 1H), 6.37 - 6.18 (m, 1H), 4.47 (tt, J = 12.2, 4.1 Hz, 1H), 4.21 (s, 2H), 4.03 (dd, J = 11.6, 4.5 Hz, 2H), 3.92 (d, J = 3.8 Hz, 2H), 3.46 (qd, J = 8.1, 4.3 Hz, 4H), 3.32 (s, 3H), 3.21 (dd, J = 11.1, 5.6 Hz, 3H), 2.78 - 2.61 (m, 2H), 2.18 (d, J = 6.7 Hz, 3H), 1.71 - 1.66 (m, 1H), 1.21 (s, 1H), 0.85 - 0.75 (m, 1H). 13 C NMR (101 MHz, CDC13) δ 172.12, 170.41, 164.18, 159.62, 156.80, 154.72, 154.33, 151.70, 149.25, 144.39, 137.80, 133.41, 132.63, 131.66, 130.78, 130.58, 129.70, 128.49, 128.15, 127.28, 126.09, 123.96, 117.68, 115.92, 107.22, 105.17, 76.38, 76.06, 75.74, 66.40, 48.98, 45.70, 44.05, 41.04, 38.18, 36.64, 35.36, 29.72, 28.91, 28.67, 27.86, 26.29, 23.54, 21.56, 16.29, 0.00. MS (ESI) m / z (M + H) + : calculated for C 47 H 53 FN 10 O7: 889.4156, found: 889.4166.

[0114] Example 2

[0115] In the preparation of the compound of this example, compound (B) of step 1) was replaced by 4-bromobutyric acid methyl ester, compound (M) of step 7) was replaced by 2-aminobutyric acid, and the remaining steps were the same as in Example 1 to obtain the compound of this example, N-(1-(4-(2-fluoro-5-((4-oxo-3,4-dihydropyrrolo[1,2-a]pyrazin-1-yl)methyl)benzoyl)piperazin-1-yl)-1-oxobutan-2-yl)-4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)butanamide, white solid product 0.027 g, yield 34.56%, which is recorded as compound LSQ-2. The structure is:

[0116]

[0117] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum and mass spectrum of compound LSQ-2 are as follows: 1 H NMR (400 MHz, CDCl3) δ 7.94-7.74 (m, 2H), 7.74-7.57 (m, 2H), 7.25 (s, 1H), 7.03-6.72 (m, 3H), 6.56-6.31 (m, 1H), 4.57-4.39 (m, 1H), 4.21 (s, 1H), 4.06 (qd, J = 13.4, 8.5 Hz, 2H), 3.94 (dt, J = 10.3, 5.5 Hz, 2H), 3.62 (d, J = 16.4 Hz, 2H), 3.57-3.38 (m, 3H), 3.38-3.24 (m, 3H), 2.77-2.67 (m, 2H), 2.45 (dt, J = 21.6, 6.0 Hz, 2H), 2.20 (s, 3H), 2.09-1.96 (m, 2H), 1.67 (t, J = 11.8 Hz, 2H), 1.51 (s, 1H), 0.92-0.70 (m, 2H). 13C NMR (101 MHz, CDC13) δ 173.86, 172.77, 170.79, 165.14, 160.62, 157.83, 157.75, 155.72, 155.36, 155.25, 152.72, 150.27, 150.07, 145.38, 138.86, 134.48, 133.61, 132.96, 132.63, 131.57, 130.71, 130.62, 129.51, 129.24, 128.31, 127.12, 124.99, 123.43, 118.69, 118.06, 116.93, 116.77, 116.29, 108.30, 106.66, 106.20, 106.12, 77.43, 77.11, 76.79, 67.40, 67.34, 66.69, 53.16, 52.18, 51.61, 50.03, 49.57, 47.28, 46.74, 45.33, 42.23, 41.91, 37.69, 32.96, 32.86, 30.63, 30.04, 29.92, 27.34, 27.29, 25.93, 25.70, 25.57, 24.77, 17.31, 17.27, 9.84, 9.76, 9.70, 1.03. MS (ESI) m / z (M+H) + : calculated for C 46 H 51 FN 10 O7: 875.3999, found: 875.4022.

[0118] Example 3

[0119] In the preparation of the compound of this example, compound (B) of Step 1) was replaced with 4-bromobutyric acid methyl ester, and compound (M) of Step 7) was replaced with 3-aminopropanoic acid, and the remaining steps were the same as in Example 1, to obtain the compound of this example, N-(3-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophtalazin-l-yl)methyl)benzoyl)piperazin-l-yl)-3-oxopropyl)-4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)butanamide, white solid product 0.030 g, yield 12.5%, which is denoted as compound LSQ-3. The structure is:

[0120]

[0121] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-3 are as follows: 1H NMR (400 MHz, CDC13) δ 8.45 - 8.25 (m, 1H), 7.87 - 7.76 (m, 2H), 7.71 - 7.59 (m, 2H), 7.25 (dt, J = 6.3, 3.6 Hz, 2H), 6.95 (td, J = 7.9, 4.6 Hz, 2H), 6.93 - 6.79 (m, 2H), 6.48 - 6.32 (m, 1H), 4.54 - 4.42 (m, 1H), 4.21 (s, 2H), 4.04 (ddd, J = 11.7, 5.7, 3.9 Hz, 2H), 3.95 - 3.82 (m, 2H), 3.63 (td, J = 21.9, 12.9 Hz, 3H), 3.45 (q, J = 11.4 Hz, 6H), 3.31 (d, J = 7.6 Hz, 4H), 3.23 - 3.12 (m, 1H), 2.70 (qd, J = 12.3, 4.6 Hz, 2H), 2.45 (dq, J = 24.4, 7.0 Hz, 2H), 2.36 - 2.26 (m, 2H), 2.19 (t, J = 3.6 Hz, 3H), 2.01 (p, J = 5.0 Hz, 2H), 1.67 (d, J = 11.8 Hz, 2H), 1.18 (t, J = 7.1 Hz, 1H). 13 C NMR (101 MHz, CDC13) δ 172.22, 172.04, 169.24, 169.06, 164.17, 164.04, 159.74, 156.79, 154.69, 154.35, 154.22, 151.70, 149.15, 144.43, 137.80, 137.73, 133.48, 132.62, 131.75, 131.54, 130.81, 130.57, 129.68, 129.63, 128.50, 128.20, 127.23, 126.06, 123.96, 117.36, 115.85, 115.81, 115.28, 115.06, 106.12, 106.06, 105.45, 105.25, 76.44, 76.32, 76.12, 75.88, 75.80, 66.39, 65.82, 65.71, 49.02, 43.81, 41.00, 40.80, 40.59, 36.59, 34.17, 34.01, 32.22, 32.07, 31.84, 28.99, 28.90, 26.32, 26.27, 24.42, 16.29, 16.24, 13.18, -0.00. MS (ESI) m / z (M+H)+: calculated for C 45 H 49 FN 10O7: 861.3843, found: 861.3849.

[0122] Example 4

[0123] In the preparation of the compound of this example, compound (B) of Step 1) was replaced with 7-bromoheptanoic acid methyl ester, and compound (M) of Step 7) was replaced with 7-aminoheptanoic acid, and the remaining steps were the same as in Example 1, to obtain the compound of this example, N-(7-(4-(2-fluoro-5-((4-oxo-3,4-dihydropyrrolo[1,2-a]pyrazin-1-yl)methyl)benzoyl)piperazin-1-yl)-7-oxoheptyl)-7-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)heptanamide, white solid product 0.019 g, yield 10.1%, which is denoted as compound LSQ-4. The structure is:

[0124]

[0125] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-4 are as follows: 1 H NMR (400 MHz, CDC13) δ 8.48 - 8.22 (m, 1H), 7.83 (s, 1H), 7.75 (d, J = 2.6 Hz, 1H), 7.71 - 7.62 (m, 2H), 7.26 (dd, J = 9.2, 4.3 Hz, 2H), 6.98 (dd, J = 8.9, 4.0 Hz, 2H), 6.78 (s, 1H), 4.47 (s, 1H), 4.22 (s, 2H), 4.03 (dd, J = 11.6, 4.5 Hz, 2H), 3.89 (t, J = 6.3 Hz, 2H), 3.62 (d, J = 41.1 Hz, 3H), 3.54 - 3.37 (m, 4H), 3.32 (s, 4H), 3.14 (q, J = 6.6 Hz, 2H), 2.69 (dd, J = 12.5, 4.6 Hz, 2H), 2.28 (t, J = 7.5 Hz, 1H), 2.20 (s, 4H), 2.09 (t, J = 7.6 Hz, 2H), 1.66 (ddd, J = 19.3, 14.8, 10.9 Hz, 5H), 1.60 - 1.45 (m, 4H), 1.40 (q, J = 7.4 Hz, 4H), 1.33 - 1.20 (m, 6H). 13C NMR (101 MHz, CDC13) δ 160.69, 158.01, 155.42, 152.74, 150.30, 138.79, 134.49, 133.66, 132.65, 131.62, 130.70, 129.53, 128.31, 127.14, 125.03, 118.66, 116.95, 108.38, 106.34, 77.42, 77.30, 77.10, 76.78, 67.82, 67.45, 50.02, 39.27, 36.69, 29.91, 29.43, 29.25, 29.01, 28.90, 27.31, 26.63, 25.86, 25.73, 24.96, 17.33. MS (ESI) m / z (M+H) + : calculated for C 52 H 63 FN 10 O7: 959.4938, found: 959.4967.

[0126] Example 5

[0127] In the preparation of the compound of this example, compound (B) of Step 1) was replaced with methyl 3-(bromomethyl)benzoate, and compound (M) of Step 7) was replaced with glycine, and the remaining steps were the same as in Example 1, to obtain the compound of this example, N-(2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophtalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-2-oxoethyl)-3-((4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)methyl)benzamide, white solid product 0.021 g, yield 10.7%, which is denoted as compound LSQ-5. The structure is:

[0128]

[0129] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-5 are as follows: 1H NMR (400 MHz, CDC13) δ 10.91 (d, J = 12.0 Hz, 1H), 8.50 - 8.40 (m, 1H), 8.00 - 7.90 (m, 2H), 7.89 (s, 1H), 7.81 - 7.73 (m, 3H), 7.73 - 7.67 (m, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.48 - 7.38 (m, 2H), 7.34 (d, J = 4.5 Hz, 2H), 7.09 - 6.99 (m, 2H), 6.82 (s, 1H), 6.59 (dd, J = 8.3, 2.6 Hz, 1H), 5.11 (s, 2H), 4.53 (tt, J = 12.2, 4.0 Hz, 1H), 4.33 - 4.19 (m, 4H), 4.07 (dd, J = 11.5, 4.2 Hz, 2H), 3.65 - 3.54 (m, 3H), 3.54 - 3.43 (m, 3H), 3.39 (s, 3H), 2.77 (qd, J = 12.5, 4.5 Hz, 2H), 2.28 (s, 3H), 1.75 - 1.62 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 167.20, 167.03, 160.47, 157.50, 155.27, 152.74, 150.28, 145.45, 138.90, 138.04, 133.71, 132.64, 131.68, 130.78, 129.51, 128.87, 128.33, 127.20, 126.66, 126.31, 124.99, 119.02, 117.02, 116.35, 116.13, 108.51, 106.44, 77.37, 77.26, 77.05, 76.73, 69.61, 67.38, 50.02, 41.74, 37.65, 29.93, 27.32, 17.36. MS (ESI) m / z (M + H) + : calculated for C 48 H 47 FN 10 O7: 895.3686, found: 895.3693.

[0130] Example 6

[0131] In the preparation of the compound of the present example, compound (B) of step 1) was replaced with methyl 3-(bromomethyl)benzoate, compound (M) of step 7) was replaced with 3-aminopropanoic acid, and the remaining steps were the same as in Example 1, to obtain the compound of the present example, N-(3-(4-(2-fluoro-5-((4-oxo-3,4-dihydropyrrolo[1,2-a]pyrazin-1-yl)methyl)benzoyl)piperazin-1-yl)-3-oxopropyl)-3-((4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)methyl)benzamide, white solid product 0.027 g, yield 11.6%, which is designated as compound LSQ-6. The structure is:

[0132]

[0133] The nuclear magnetic resonance spectrum of hydrogen, the nuclear magnetic resonance spectrum of carbon, and the mass spectrum of compound LSQ-6 are as follows: 1 H NMR (400 MHz, CDCl3) δ 11.11 (d, J = 9.2 Hz, 1H), 8.48-8.35 (m, 1H), 7.98-7.82 (m, 3H), 7.72 (ddd, J = 16.9, 6.2, 3.0 Hz, 4H), 7.59 (d, J = 7.6 Hz, 1H), 7.40 (dt, J = 13.2, 7.0 Hz, 2H), 7.34-7.27 (m, 2H), 7.08-6.97 (m, 2H), 6.87 (s, 1H), 6.58 (dd, J = 8.3, 2.5 Hz, 1H), 5.09 (s, 2H), 4.52 (ddt, J = 12.1, 8.0, 3.9 Hz, 1H), 4.27 (s, 2H), 4.07 (dd, J = 11.5, 4.0 Hz, 2H), 3.80-3.65 (m, 4H), 3.64 (s, 1H), 3.60-3.41 (m, 6H), 3.39 (s, 3H), 3.28 (s, 2H), 2.81-2.56 (m, 4H), 2.28 (s, 3H), 1.72 (d, J = 10.0 Hz, 2H). 13C NMR (101 MHz, CDC13) δ 169.46, 169.26, 166.36, 166.27, 164.16, 164.02, 159.48, 156.47, 154.23, 151.69, 149.23, 144.40, 137.87, 136.84, 133.59, 133.42, 132.63, 131.63, 130.72, 130.60, 129.73, 128.47, 128.19, 127.74, 127.27, 126.12, 125.60, 125.22, 123.95, 118.03, 115.98, 115.28, 115.06, 107.42, 105.45, 76.34, 76.23, 76.02, 75.71, 68.66, 66.34, 48.97, 45.90, 45.62, 44.44, 43.88, 40.97, 40.82, 40.66, 40.17, 36.63, 34.60, 31.87, 28.90, 26.29, 16.32, -0.00. MS (ESI) m / z (M+H) + : calculated for C 49 H 49 FN 10 O7: 909.3843, found: 909.3816.

[0134] Example 7

[0135] In the preparation of the compound of this example, compound (B) of step 1) was prepared using methyl 4-(bromoethyl)benzoate, compound (R) of step 9) was reacted with compound (L) of step 13) to perform step 14), and the remaining steps were the same as in Example 1, to obtain the compound of this example, 4-(4-fluoro-3-(4-(4-((4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)methyl)benzoyl)piperazin-1- carbonyl)benzyl)phthalazine-1(2H)-one, white solid product 0.018 g, yield 23.04%, which is denoted as compound LSQ-7. The structure is:

[0136]

[0137] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-7 are as follows: 1H NMR (400 MHz, CDC13) δ 8.28 (s, 2H), 8.06 (s, 1H), 7.94 (d, J = 7.9 Hz, 2H), 7.85 (s, 1H), 7.59 - 7.35 (m, 5H), 7.29 (d, J = 49.1 Hz, 1H), 7.06 (d, J = 8.4 Hz, 1H), 6.63 (dd, J = 8.4, 2.6 Hz, 1H), 5.12 (d, J = 15.5 Hz, 2H), 4.36 (d, J = 30.9 Hz, 2H), 3.92 (dd, J = 11.6, 4.3 Hz, 2H), 3.65 (s, 3H), 3.30 (s, 3H), 2.61 - 2.52 (m, 2H), 2.17 (s, 3H), 1.66 (dd, J = 12.7, 4.0 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 159.85, 156.87, 155.97, 152.48, 150.08, 139.89, 135.28, 133.99, 131.02, 129.88, 127.94, 127.69, 125.93, 122.60, 116.95, 109.59, 69.03, 66.98, 49.60, 40.57, 40.37, 40.16, 39.95, 39.74, 39.53, 39.32, 29.95, 27.55, 17.77. MS (ESI) m / z (M+H) + : calculated for C 46 H 44 FN9O6: 838.3472, found: 838.3455.

[0138] Example 8

[0139] In the preparation of the compound of this example, step 7) compound (M) uses 5- aminopentanoic acid, and the remaining steps are the same as in Example 1, to obtain the compound of this example, N-(5-(4-(2-fluoro-5-((4-oxo-3,4-dihydropyrrolo[1,2-a]pyrazin-1- yl)methyl)benzoyl)piperazin-1-yl)-5-oxopentyl)-5-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro- 2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)pentanamide, white solid product 0.021 g, yield 26.88%, recorded as compound LSQ-8. The structure is:

[0140]

[0141] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum and mass spectrum of compound LSQ-8 are as follows: 1H NMR (400 MHz, CDC13) δ 8.44 (d, J = 7.1 Hz, 1H), 7.90 (s, 1H), 7.75 (q, J = 11.3 Hz, 4H), 7.32 (p, J = 7.8 Hz, 3H), 7.04 (d, J = 9.0 Hz, 2H), 6.51 (d, J = 8.8 Hz, 2H), 4.53 (s, 1H), 4.28 (s, 2H), 4.13-4.05 (m, 2H), 4.03-3.91 (m, 2H), 3.84-3.61 (m, 3H), 3.60-3.46 (m, 4H), 3.41 (d, J = 14.7 Hz, 4H), 3.34-3.14 (m, 4H), 2.83-2.66 (m, 2H), 2.37 (td, J = 17.4, 8.1 Hz, 2H), 2.26 (s, 5H), 1.77 (q, J = 15.8 Hz, 7H), 1.67-1.63 (m, 1H), 1.59-1.46 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 173.40, 171.67, 165.27, 160.77, 158.17, 157.77, 155.71, 155.19, 152.70, 150.46, 145.51, 142.06, 138.56, 134.46, 133.66, 132.04, 131.84, 131.60, 130.74, 129.49, 128.20, 127.04, 126.46, 125.10, 124.99, 119.19, 118.19, 116.86, 116.31, 110.47, 108.57, 106.64, 77.46, 77.15, 76.83, 67.39, 51.60, 50.07, 47.10, 46.75, 45.63, 45.06, 42.16, 41.94, 41.72, 41.21, 38.98, 38.79, 37.63, 36.28, 33.46, 32.43, 29.91, 28.84, 27.34, 22.58, 22.05, 21.72, 17.31. MS (ESI) m / z (M + H) + : calculated for C 48 H 55 FN 10 O7: 903.4312, found: 903.4315.

[0142] Example 9

[0143] In the preparation of the compound of this example, compound (B) of step 1) was replaced with 6-bromohexanoic acid methyl ester, and the remaining steps were the same as in Example 1 to obtain the compound of this example, N-(4-(4-(2-fluoro-5-((4-oxo-3,4-dihydropyrrolo[1,2-a]pyrazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)-6-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)hexanamide, white solid product 0.024 g, yield 30.72%, which is denoted as compound LSQ-9. The structure is:

[0144]

[0145] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-9 are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.42-8.33 (m, 1H), 7.84 (s, 1H), 7.74 (d, J = 2.6 Hz, 1H), 7.69 (tq, J = 8.8, 4.4 Hz, 2H), 7.65 (t, J = 3.7 Hz, 1H), 7.25 (dd, J = 8.2, 4.8 Hz, 2H), 7.00-6.91 (m, 2H), 6.79 (s, 1H), 6.44 (dt, J = 8.7, 2.4 Hz, 1H), 6.14 (d, J = 20.9 Hz, 1H), 4.47 (t, J = 4.1 Hz, 1H), 4.21 (s, 2H), 4.04 (dd, J = 11.6, 4.4 Hz, 2H), 3.91 (tt, J = 6.3, 2.8 Hz, 2H), 3.67 (s, 2H), 3.58 (s, 2H), 3.48 (dt, J = 12.3, 3.0 Hz, 3H), 3.42 (d, J = 4.2 Hz, 1H), 3.33 (s, 3H), 3.24-3.18 (m, 3H), 2.69 (dd, J = 12.5, 4.6 Hz, 1H), 2.35 (t, J = 6.6 Hz, 1H), 2.20 (s, 2H), 2.12 (t, J = 7.5 Hz, 2H), 1.78 (d, J = 6.2 Hz, 2H), 1.68 (d, J = 3.7 Hz, 1H), 1.65 (d, J = 3.5 Hz, 1H), 1.44 (d, J = 7.8 Hz, 1H), 1.28 (d, J = 15.6 Hz, 1H), 1.18 (s, 2H). 13CNMR (101 MHz, CDC13) δ 171.47, 160.49, 157.93, 155.42, 152.76, 150.31, 145.46, 138.80, 134.42, 133.70, 132.67, 131.67, 130.73, 129.53, 128.34, 127.18, 125.01, 118.76, 116.96, 108.35, 106.34, 77.38, 77.27, 77.07, 76.86, 76.75, 70.53, 67.63, 67.46, 50.04, 45.11, 39.20, 37.68, 36.70, 31.94, 30.91, 29.94, 29.71, 29.08, 27.32, 25.84, 25.51, 24.56, 17.33. MS (ESI) m / z (M+H) + : calculated for C 48 H 55 FN 10 O7: 903.4312, found: 903.4315.

[0146] Example 10

[0147] In the preparation of the compound of this example, compound (B) of step 1) was replaced by 4-bromobutyric acid methyl ester, and compound (M) of step 7) was replaced by 3-azetidinecarboxylic acid, and the remaining steps were the same as in Example 1, to obtain the compound of this example, 4-(4-fluoro-3-(4-(1-(4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)butanoyl)azetidin-3-carbonyl)piperazin-1- carbonyl)benzyl)phthalazine-1(2H)-one, white solid product 0.029 g, yield 37.12%, recorded as compound LSQ-10. The structure is:

[0148]

[0149] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum and mass spectrum of compound LSQ-10 are as follows: 1H NMR (400 MHz, CDC13) δ 8.41 - 8.31 (m, 1H), 7.83 (q, J = 4.7 Hz, 1H), 7.75 (dd, J = 8.9, 2.6 Hz, 1H), 7.72 - 7.61 (m, 3H), 7.26 (dt, J = 10.3, 5.1 Hz, 2H), 7.00 - 6.90 (m, 2H), 6.86 (d, J = 5.3 Hz, 1H), 6.44 (dd, J = 8.3, 2.9 Hz, 1H), 4.46 (tt, J = 13.2, 4.7 Hz, 2H), 4.36 (d, J = 7.0 Hz, 1H), 4.21 (s, 2H), 4.17 - 4.08 (m, 1H), 4.03 (dd, J = 9.4, 5.3 Hz, 2H), 3.97 - 3.90 (m, 2H), 3.88 (s, 1H), 3.75 - 3.50 (m, 4H), 3.44 (dd, J = 11.3, 7.4 Hz, 2H), 3.33 - 3.30 (m, 3H), 3.26 - 3.04 (m, 3H), 2.74 - 2.64 (m, 2H), 2.49 (d, J = 7.4 Hz, 1H), 2.36 (d, J = 7.2 Hz, 1H), 2.19 (q, J = 4.2 Hz, 3H), 2.06 - 1.93 (m, 3H), 1.79 - 1.58 (m, 3H). 13 C NMR (101 MHz, CDC13) δ 171.90, 168.55, 164.20, 159.72, 156.74, 156.69, 154.33, 154.27, 151.69, 149.31, 144.44, 137.80, 137.71, 133.52, 132.64, 131.46, 130.94, 130.60, 129.74, 128.48, 127.21, 126.07, 123.94, 117.85, 117.77, 115.95, 107.33, 105.45, 105.05, 76.41, 76.29, 76.09, 75.87, 75.77, 66.37, 65.82, 65.69, 50.76, 49.20, 48.99, 43.73, 41.04, 40.96, 36.61, 29.65, 29.05, 28.89, 26.64, 26.35, 26.29, 23.56, 16.28, 16.24, 0.00. MS (ESI) m / z (M + H) + : calculated for C 46 H 49 FN 10 O7:873.3843, found:873.3815.

[0150] Comparative Example 1

[0151] In the preparation of the compound of this example, compound (B) of Step 1) was replaced with 7-bromo-2,2-dimethylheptanoate, compound (M) of Step 7) was replaced with 6-aminohexanoic acid, and the remaining steps were the same as in Example 1, to obtain the compound of this example, N-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazine-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexyl)-2,2-dimethyl-7-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)heptanamide, white solid product 0.013 g, yield 16.64%, which is designated as compound LSQ-11. The structure is:

[0152]

[0153] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-11 are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.39 (dd, J = 7.0, 2.1 Hz, 1H), 7.83 (s, 1H), 7.77-7.56 (m, 3H), 7.30-7.22 (m, 1H), 6.98 (dd, J = 8.2, 4.9 Hz, 2H), 6.79 (s, 1H), 6.47-6.33 (m, 1H), 5.76 (d, J = 5.5 Hz, 1H), 3.32 (s, 3H), 2.69 (dd, J = 12.5, 4.4 Hz, 2H), 1.08 (d, J = 2.3 Hz, 5H). 13C NMR (101 MHz, CDC13) δ 177.75, 171.60, 165.22, 160.52, 157.97, 155.74, 155.42, 152.74, 150.31, 145.43, 138.78, 134.43, 134.40, 133.66, 132.62, 131.77, 131.70, 131.62, 130.70, 129.52, 129.16, 128.33, 127.16, 125.01, 118.75, 116.99, 116.96, 116.12, 108.44, 106.47, 106.42, 77.38, 77.27, 77.07, 76.75, 67.88, 67.44, 50.02, 46.82, 45.09, 42.19, 42.02, 41.67, 41.26, 39.12, 37.69, 35.53, 32.97, 29.91, 29.70, 29.35, 29.26, 27.30, 26.71, 26.66, 26.49, 26.19, 25.50, 24.73, 24.44, 17.33, 1.04. MS (ESI) m / z (M+H) + : calculated for C 50 H 59 FN 10 O7: 931.4625, found: 931.4655.

[0154] Comparative Example 2

[0155] In the preparation of the compound of this example, compound (B) of Step 1) was replaced with 7-bromo-2,2-dimethylheptanoate, and compound (M) of Step 7) was replaced with 3-azetidinecarboxylic acid, and the remaining steps were the same as in Example 1, to obtain the compound of this example, 4-(3-(4-(1-(2,2-dimethyl-7-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)heptanoyl)azetidin-3- carbonyl)piperazin-1-ylcarbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one, white solid product 0.016 g, yield 20.48%, which is denoted as compound LSQ-12. The structure is:

[0156]

[0157] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-12 are as follows: 1H NMR (400 MHz, CDC13) δ 8.38 (dt, J = 7.1, 3.0 Hz, 1H), 7.88 - 7.79 (m, 1H), 7.78 - 7.60 (m, 4H), 7.26 (dq, J = 9.0, 3.2 Hz, 2H), 6.98 (td, J = 8.7, 3.9 Hz, 2H), 6.80 (d, J = 8.5 Hz, 1H), 6.46 (dt, J = 8.2, 2.4 Hz, 1H), 4.47 (tt, J = 12.3, 4.1 Hz, 1H), 4.21 (d, J = 7.6 Hz, 2H), 4.03 (dd, J = 11.8, 4.3 Hz, 3H), 3.89 (td, J = 6.6, 2.4 Hz, 3H), 3.57 (s, 3H), 3.50 - 3.39 (m, 4H), 3.32 (d, J = 2.7 Hz, 4H), 3.27 - 3.07 (m, 3H), 2.69 (qd, J = 12.4, 4.3 Hz, 2H), 2.19 (d, J = 2.8 Hz, 3H), 1.68 (ddd, J = 24.1, 10.5, 5.3 Hz, 5H), 1.48 - 1.34 (m, 4H), 1.27 - 1.16 (m, 4H), 1.13 - 1.02 (m, 6H). 13 C NMR (101 MHz, CDC13) δ 176.29, 168.79, 168.59, 164.20, 159.70, 156.98, 154.69, 154.36, 151.70, 149.28, 144.40, 137.74, 133.51, 132.64, 131.56, 130.91, 130.83, 130.60, 129.67, 128.48, 128.19, 127.24, 126.10, 123.94, 122.40, 117.72, 115.94, 115.31, 115.05, 107.49, 105.36, 76.39, 76.28, 76.08, 75.76, 66.80, 66.40, 53.26, 48.98, 45.94, 45.64, 44.17, 43.73, 41.14, 41.07, 41.01, 40.77, 40.54, 39.26, 36.63, 30.30, 28.87, 28.66, 28.19, 26.28, 25.62, 24.51, 24.25, 23.55, 16.29, 16.26, -0.00. MS (ESI) m / z (M + H) + : calculated for C 51 H 59 FN 10 O7: 943.4625, found: 943.4612.

[0158] Comparative Example 3

[0159] In the preparation of the compound of this example, compound (B) of Step 1) was replaced with methyl 4-(bromomethyl)benzoate, and compound (M) of Step 7) was replaced with 7-aminoheptanoic acid, and the remaining steps were the same as in Example 1, to obtain the compound of this example, N-(6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazine-1-yl)methyl)benzoyl)piperazin-1-yl)-6-oxohexyl)-4-(4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)phenoxy)benzamide, white solid product 0.019 g, yield 24.32%, which is designated as compound LSQ-13. The structure is:

[0160]

[0161] The nuclear magnetic resonance hydrogen spectrum, nuclear magnetic resonance carbon spectrum, and mass spectrum of compound LSQ-13 are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.50-8.40 (m, 1H), 7.96 (d, J = 2.7 Hz, 1H), 7.88 (s, 1H), 7.80-7.71 (m, 2H), 7.71-7.64 (m, 1H), 7.47-7.42 (m, 2H), 7.31 (q, J = 7.3 Hz, 3H), 7.06 (dd, J = 13.4, 8.5 Hz, 3H), 6.89-6.79 (m, 1H), 6.59 (dd, J = 8.4, 2.5 Hz, 1H), 4.53 (tt, J = 12.4, 4.2 Hz, 1H), 4.27 (s, 2H), 4.06 (dd, J = 11.6, 4.5 Hz, 3H), 3.71 (d, J = 4.2 Hz, 4H), 3.64 (d, J = 3.1 Hz, 2H), 3.59-3.44 (m, 8H), 3.39 (s, 3H), 3.17 (t, J = 6.6 Hz, 3H), 2.75 (s, 3H), 2.28 (s, 3H), 2.16-1.95 (m, 4H), 1.72 (dd, J = 13.0, 4.1 Hz, 2H), 1.58 (d, J = 8.0 Hz, 3H), 1.42 (s, 1H). 13C NMR (101 MHz, CDC13) δ 160.52, 157.63, 155.30, 152.73, 150.31, 145.44, 138.89, 136.47, 134.66, 134.40, 133.66, 132.58, 131.63, 130.76, 129.61, 129.52, 128.33, 127.16, 125.01, 119.10, 117.04, 108.51, 106.52, 77.39, 77.07, 76.76, 69.72, 67.39, 50.05, 43.58, 42.20, 39.52, 37.70, 33.06, 29.94, 29.24, 28.84, 27.33, 26.52, 24.93, 17.36. MS (ESI) m / z (M+H) + : calculated for C 46 H 49 FN 10 O7: 979.4625, found: 979.4606.

[0162] Performance test

[0163] I. Test of DNA-PK inhibitory activity of compounds

[0164] The enzyme inhibitory activity of compounds was determined by fluorescence analysis, in which DNA-PK0 (V4106) enzyme, ADP-Glo TM Kinase Assay (V9101) and ATP (V915B) were purchased from Promega, 384 well small volume white plate (784075) was purchased from Greiner, and Envision was purchased from PerkinElmer. The specific test method is as follows:

[0165] The experimental process is as follows:

[0166] 1. Thaw DNA-PK enzyme, DNA-Dependent Protein Kinase Substrate, 5X Reaction Buffer A, DNA-PK Activation Buffer, DTT (100 mM) and ATP (10 mM) on ice, and the above reagents need to be placed on ice throughout the experiment;

[0167] 2. 5x Reaction Buffer A and 10x DNA-PK Activation Buffer were prepared with deionized water to 1x buffer, and DTT was added to the 1x buffer, the concentration of DTT in the 1x buffer was 50 μM;

[0168] 3. The test compound stock solution was prepared with 1x buffer to 5 times the final concentration gradient, 1 μl / well of test compound was added to the white microplate, at this time the DMSO concentration of the compound could not be more than 5%, the final DMSO concentration of the experiment could not be more than 1%, and the microplate was centrifuged at 1000 rpm for 1 minute on the centrifuge;

[0169] Positive control well (Pos. Ctrl): 1 μL / well of 1x buffer containing 5% DMSO;

[0170] Blank control well (Blank): 1 μL / well of 1x buffer containing 5% DMSO;

[0171] 4. After the DNA-PK enzyme was completely thawed, the DNA-PK enzyme was diluted to 2.5 unit / μL using 1x buffer, and 2 μL / well was added to the white microplate, at this time the amount of DNA-PK enzyme in each well was 5 units; 2 μL / well of 1x buffer was added to the blank control well; this step was performed on ice, and after addition, the microplate was centrifuged at 1000 rpm for 1 minute on the centrifuge;

[0172] 5. Prepare DNA-PK peptide substrate / ATP mixture:

[0173] DNA-PK peptide substrate / ATP mixture: 1x buffer was used to dilute the DNA-PK peptide substrate 20 times, and ATP (10 mM) was diluted 80 times, at this time the concentration of DNA-PK peptide substrate was 0.5 ug / ul, and the concentration of ATP was 125 μM. This step was performed on ice;

[0174] 6. 2 μL / well of DNA-PK peptide substrate / ATP mixture was added to the white microplate, at this time the concentration of DNA-PK peptide substrate was 0.2 ug / ul, and the concentration of ATP was 50 μM, and after addition the microplate was centrifuged at 1000 rpm for 1 minute;

[0175] 7 After centrifugation, the microplate was covered with film, the film was pressed tightly, and incubated at 25°C for 1 hour;

[0176] 8. After the end of the incubation, chemiluminescence detection was performed using Envision, and the luminescence value (RLU) was read;

[0177] 9. Inhibition rate calculation:

[0178] %Inhibtion = (RLU (Sample) - RLU (Pos. Ctrl)) / (RLU (Blank) - RLU (Pos. Ctrl)) x 100%

[0179] The results are shown in Table 1.

[0180] Table 1 Test results of compounds on DNA-PK inhibition activity

[0181] Compound DNA-PK IC 50 (nM)]]> LSQ-1 10.84 LSQ-2 23.21 LSQ-3 15.04 LSQ-4 1.99 LSQ-5 1.98 LSQ-6 4.51 LSQ-7 8.28 LSQ-8 8.38 LSQ-9 29.58 LSQ-10 27.04 LSQ-11 ＞50 LSQ-12 ＞50 LSQ-13 ＞50 Wortmannin 15.35

[0182] wherein the positive control Wortmannin was compared with samples LSQ-1-10, respectively. The experimental results of Table 1 show that compounds LSQ-1-LSQ-10 have strong inhibition effect on DNA-PK, and the activity is comparable to that of Wortmannin. Compounds LSQ-11-LSQ-13 have poor inhibition effect.

[0183] II. Test of PARP1 inhibition activity of compounds

[0184] The enzyme inhibition activity of the compounds was determined by fluorescence analysis, wherein the PARP1 Chemiluminescent Assay Kit (80569) was purchased from BPS Company, and the Envision was purchased from PerkinElmer Company. The specific test method is as follows:

[0185] The experimental process is as follows:

[0186] 1. One day before the experiment, thaw 5x histone mixture on ice;

[0187] 2. Dilute 5x histone mixture 5 times with 1x PBS to prepare 1x histone mixture;

[0188] 3. Take 25 μL / well of 1x histone mixture and add it to the test plate, which is centrifuged at 1000 rpm for 1 minute on a centrifuge; incubate at 4°C overnight;

[0189] 4. After the end of the incubation, spin dry the liquid in the test plate, and take 100 μL / well of PBST buffer to wash the test plate 3 times;

[0190] 5. Add 100 μL / well of Blocking buffer 3 to the assay plate and incubate at 25°C for 60-90 minutes;

[0191] 6. Dilute 10x PARP Buffer 10-fold with deionized water and place on ice until use;

[0192] 7. After the incubation is complete, flick the liquid from the assay plate and repeat the plate washing 3 times;

[0193] 8. Take 2.5 μL / well of 10-fold final concentration gradient of compound working solution and add to the well plate according to the experimental layout;

[0194] Positive control wells (Pos. Ctrl): 2.5 μL / well of IX PARP buffer containing 10% DMSO;

[0195] Blank control wells (Blank): 2.5 μL / well of IX PARP buffer containing 10% DMSO;

[0196] 9. After the enzyme is completely dissolved, dilute the enzyme stock solution to 2 ng / μL with IX PARP buffer, take 10 μL / well of enzyme solution and add to the assay plate, at this time the enzyme amount of PARP1 in each well is 20 ng / well; add 10 μL / well of IX PARP buffer to the blank control wells; this step is performed on ice, after adding, centrifuge at 1000 rpm for 1 minute;

[0197] 10. Add 12.5 μL of master mixture to each well of the PARP assay plate, 12.5 μL of master mixture includes 1.25 μL of 10x PARP buffer, 1.25 μL of 10x PARP Assay mixture, 2.5 μL of Activated DNA (5x), 7.5 μL of water; seal the assay plate and incubate at 25°C for 60 minutes;

[0198] 11. After the incubation is complete, flick the liquid from the assay plate and repeat the plate washing 3 times;

[0199] 12. Dilute Streptavidin-HRP in the kit 50-fold with Blocking buffer solution, take 25 μL / well and add to the assay plate, incubate at 25°C for 30 minutes;

[0200] 13. After the incubation is complete, flick the liquid from the assay plate and repeat the plate washing 3 times;

[0201] 14. Mix HRP chemiluminescent substrate A and HRP chemiluminescent substrate B in a 1:1 ratio, add 50 μL / well of the mixture to the test plate, and immediately perform Luminescence detection using Envision to read the luminescence value (RLU);

[0202] 15. Inhibition rate calculation:

[0203] % Inhibtion = (RLU (Sample) - RLU (Pos. Ctrl)) / (RLU (Blank) - RLU (Pos. Ctrl)) x 100%

[0204] The results are shown in Table 2.

[0205] Table 2. Test results of PARP1 inhibition activity of compounds

[0206]

[0207]

[0208] Positive control Niraparib was compared with sample LSQ-1-10. The experimental results in Table 2 show that compounds LSQ-1 to LSQ-10 have strong inhibitory effect on PARP1, and their activity is comparable to Niraparib. Compounds LSQ-11 to LSQ-13 have poor inhibitory effect.

[0209] III. Test of activity of compounds on breast cancer cells

[0210] (1) Test of cytotoxicity of compounds on 4T1 cells

[0211] MTT was used to determine the cytotoxicity of the compounds, 4T1 cells were purchased from RiboBioscience, MTT was purchased from Bide Pharmatech Co., Ltd., 1640 was purchased from RiboBioscience, bovine serum solution was purchased from RiboBioscience, and penicillin streptomycin antibacterial solution was purchased from RiboBioscience. The specific test method is as follows:

[0212] The experimental process is as follows:

[0213] 1. Preparation of 4T1 cell suspension: cell counting; 2. Inoculation into 96-well plates, about 3000 cells per well, about 100ul cell suspension per well; 3. Incubation in a 37°C incubator overnight; 4. Aspirate the culture medium, and sequentially add drugs LSQ-1 to LSQ-13, and positive drugs AZD-7648 and Olaparib diluted to 25μmol into 98-well plates, with three replicates; 5. Incubation in a 37°C incubator for 48h; 6. Add 20ul MTT in the dark, and add the gun head into the culture solution, and gently tap the culture plate after adding the reagent to help mixing; 7. Incubation in a 37°C incubator for 4h; 8. Measure the absorbance at 490nm and 570nm; 9. Calculate the inhibition rate.

[0214] Table 3 Results of compound toxicity test on 4T1 cells

[0215] Compound 4T1 cell 25 μmol inhibition rate (%) LSQ-1 14 LSQ-2 44 LSQ-3 81 LSQ-4 19 LSQ-5 50 LSQ-6 54 LSQ-7 40 LSQ-8 36 LSQ-9 35 LSQ-10 24 LSQ-11 ＜10 LSQ-12 ＜10 LSQ-13 ＜10 AZD-7648 32 Olaparib 53

[0216] Comparison of positive controls AZD-7648 and Olaparib with samples LSQ-1-10. The experimental results in Table 3 show that compounds LSQ-1~LSQ-10 have strong toxicity on 4T1 cells, and most of the activity is comparable to AZD-7648. Compounds LSQ-11~LSQ-13 have poor inhibition effect.

[0217] (2) Compound toxicity test on B16-F10 cells

[0218] MTT was used to determine the cytotoxicity of the compounds, and B16-F10 cells were purchased from Hunan Biotech Co., Ltd., and MTT was purchased from Bide Pharmaceutical Co., Ltd. The B16-F10 special medium was purchased from Ponsay Biotech Co., Ltd., and the specific test method is as follows:

[0219] The experimental process is as follows:

[0220] 1. Preparation of B16-F10 cell suspension: cell counting; 2. Inoculation into 96-well plates, about 3000 cells per well, about 100ul cell suspension per well; 3. Incubation in a 37°C incubator overnight; 4. Aspirate the culture medium, and sequentially add drugs LSQ-1 to LSQ-13, and positive drugs AZD-7648 and Olaparib diluted to 25μmol into 98-well plates, with three replicates; 5. Incubation in a 37°C incubator for 48h; 6. Add 20ul MTT in the dark, and add the gun head into the culture solution, and gently tap the culture plate after adding the reagent to help mixing; 7. Incubation in a 37°C incubator for 4h; 8. Measure the absorbance at 490nm and 570nm; 9. Calculate the inhibition rate.

[0221] Table 4 Results of compound toxicity test on B16-F10 cells

[0222] Compound B16-F10 cell 25 μmol inhibition rate (%) LSQ-1 79 LSQ-2 79 LSQ-3 94 LSQ-4 80 LSQ-5 83 LSQ-6 84 LSQ-7 84 LSQ-8 75 LSQ-9 93 LSQ-10 82 LSQ-11 ＜10 LSQ-12 ＜10 LSQ-13 ＜10 AZD-7648 84 Olaparib 59

[0223] The positive control AZD-7648 and olaparib were compared with sample LSQ-1-10. The experimental results of Table 4 show that compounds LSQ-1-LSQ-10 have strong toxicity to B16-F10 cells, and the activity is comparable to AZD-7648. Compounds LSQ-11-LSQ-13 have poor inhibition effect.

[0224] The weight of the related components mentioned in the embodiment specification of the present application can not only refer to the specific content of each component, but also represent the proportional relationship between the weights of each component. Therefore, as long as the content of the related components in the embodiment specification of the present application is enlarged or reduced in proportion, it is within the scope disclosed in the embodiment specification of the present application.

Claims

1. A compound, characterized in that: The compound is: 。 2. A pharmaceutical composition, characterized by: The pharmaceutical composition comprises the compound or a pharmaceutically acceptable salt thereof as claimed in claim 1.

3. Use of the compound or a pharmaceutically acceptable salt thereof as claimed in claim 1 in the preparation of a medicament for the treatment and / or prevention and / or adjuvant therapy of cancer.

4. The use according to claim 3, wherein: The cancer is selected from at least one of breast cancer, melanoma, leukemia.

5. Use of the compound or a pharmaceutically acceptable salt thereof as claimed in claim 1 in the preparation of a DNA-PK inhibitor and / or a PARP1 inhibitor.

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