A tricyclic pyrazolopyrimidine aryl ester derivative and its use

Abstract

The present invention relates to a tricyclic pyrazolopyrimidine aryl ester derivative and its use. The derivative uses ethyl 5-amino-1-methylpyrazole-4-carboxylate as the starting material, reacts with 2-azepanone under the catalysis of phosphorus oxychloride, reacts with an equal amount of 3,4,5-trimethoxybenzaldehyde and a catalytic amount of sodium hydroxide in an ethanol solution, then refluxes with an acetic acid solution of 40% hydrobromic acid as the solvent, and finally reacts with differently substituted acyl chlorides under the action of a catalytic amount of base to obtain derivatives (D1-D22). In addition, under different conditions, compound (B) is dehydrogenated or thio-substituted to obtain compounds (E1-E3); 1,5,6-trimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one reacts with differently substituted aldehydes to obtain compounds (F1-F3); and the inhibitory activities of these 29 compounds were investigated simultaneously. The results showed that all 29 compounds had inhibitory activities against HCT116 colon cancer cells and HGC27 gastric cancer cells.

Description

Technical Field

[0001] The present invention relates to a tricyclic pyrazolopyrimidine aryl ester derivative and its use. Through cell activity screening of such compounds, the results show that 25 compounds have inhibitory activity against HCT116 colon cancer cells, and 25 compounds have inhibitory activity against HGC27 gastric cancer cells. Background Art

[0002] Cancer, commonly known as malignant tumor, is a class of diseases that seriously endanger human life and health. In recent years, the incidence and fatality rate of cancer have been on the rise. According to the latest data released by the International Agency for Research on Cancer of the World Health Organization, in 2020, the number of newly diagnosed cancer cases globally was approximately 19.3 million, and the number of deaths was approximately 10 million. Therefore, finding highly effective new anti-tumor drugs has become a very important and urgent topic in current cancer treatment research.

[0003] Anti-tumor small molecule targeted preparations have the characteristics of high targeting and remarkable efficacy, thus improving the quality of life of patients. They have now become a type that has attracted much attention in clinical medications. In the medical field, saturated heterocyclic compounds represented by piperidine and pyrimidine have long been known, and among all small molecule therapeutic drugs listed in the US FDA Orange Book, they are currently the most representative saturated heterocyclic systems. On this basis, introducing some non-pure aromatic or heteroaromatic cyclic systems such as benzene, pyrrole, pyrazole, pyrimidine, and pyridine into the molecule will always achieve some unexpected effects, including providing greater diversity (through stereoisomerism) on the basis of a slightly increased molecular weight, and at the same time, it can further enrich the novelty of the structure of such compounds. Therefore, the structure of drug molecules is gradually changing, and saturated ring systems represented by heterocycles are attracting more and more attention.

[0004] The pyrimidine or pyrimidinone fragment can form various bases due to its unique structure, so it is an important class of molecules indispensable in the field of life science. If pyrimidine is combined with a five-membered heterocycle, it will provide a wider range of nitrogen-containing groups in the process of drug design and development.

[0005] Our research group has been engaged in the synthesis of heterocyclic compounds, the anti-tumor structure-activity relationship, and the study of the anti-tumor multi-drug resistance reversal activity of natural compounds for a long time. M. Chauhan et al. replaced the thiophene ring with a pyrazole group and synthesized a series of new tricyclic pyrazolo[3,4-d]pyrimidine derivatives. It was found that pyrazolo[3,4-d]pyrimidine compounds have a variety of biological activities, such as antibacterial, anti-inflammatory, antiviral, etc. At the same time, enzyme inhibition activity screening found that pyrazolo[3,4-d]pyrimidine compounds can also be used as various kinase inhibitors, especially in the aspect of being a PDE9A inhibitor with better effects. Currently, most of the reported PDE9A inhibitors that have completed phase II clinical trials for Alzheimer's disease contain a pyrazolopyrimidone skeleton. On the other hand, the newly designed pyrazolo[3,4-d]pyrimidine compounds show good anti-tumor activity and are further studied as lead compounds for chemotherapy drugs.

[0006] References

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[0018] Our research group has been engaged in the synthesis and biological activity research of pyrazolo[3,4-d]pyrazolopyrimidinone compounds for a long time. Research has shown that pyrazolo[3,4-d]pyrazolopyrimidinone compounds have anti-tumor activity. Searching for and exploring the synthesis methods and biological activities of pyrazolo[3,4-d]pyrazolopyrimidinone compounds has great theoretical and application value for the discovery of new drugs and lead compounds. Based on the analysis of relevant patents and literature at home and abroad, combined with the previous research work of our research group, the structure of pyrazolo[3,4-d]pyrazolopyrimidinone compounds was modified, and different types of heterocycles and substituents were introduced into pyrazolo[3,4-d]pyrazolopyrimidinone compounds to improve their anti-tumor activity and drug-likeness, and the inhibitory activities of these compounds against HCT116 colon cancer cells and HGC27 gastric cancer cells were studied to discover anti-tumor candidate drugs with high efficiency and low toxicity.

[0019] In this invention, substituent modification of pyrazolo[3,4-d]pyrazolopyrimidinone compounds and the use of drug design principles were carried out to explore their anti-tumor activity. The results of activity screening showed that 29 compounds had inhibitory activity against HCT116 colon cancer cells; 29 compounds had inhibitory activity against HGC27 gastric cancer cells; Summary of the Invention

[0020] The object of the present invention is to provide a tricyclic pyrazolopyrimidine aryl ester derivative and its use. The derivative uses ethyl 5-amino-1-methylpyrazole-4-carboxylate as the starting material, reacts with 2-azepanone under the catalysis of phosphorus oxychloride to obtain 1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (A). An equal amount of 3,4,5-trimethoxybenzaldehyde and a catalytic amount of sodium hydroxide are added to an ethanol solution, and heated under reflux to obtain the monosubstituted compound (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (B). Then, reflux reaction is carried out using an acetic acid solution of 40% hydrobromic acid as the solvent to obtain (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (C). Finally, under the action of a catalytic amount of base, it reacts with differently substituted acyl chlorides to obtain the target (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one derivatives (D1-D23). In addition, compounds (E1-E3) are obtained by dehydrogenation or thio-substitution of compound (B) under different conditions. Compounds (F1-F3) are obtained by reacting 1,5,6-trimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one with different aldehydes under alkaline conditions. The antitumor activities of the above compounds were investigated. The results showed that: using doxorubicin (DOX) as the positive control drug, 29 compounds had inhibitory activities against HCT116 colon cancer cells, and 29 compounds had inhibitory activities against HGC27 gastric cancer cells.

[0021] A tricyclic pyrazolopyrimidine aryl ester derivative described in the present invention has the following structure:

[0022]

[0023] Wherein:

[0024] Compound D1 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pentanoate;

[0025] Compound D2 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl dodecanoate;

[0026] Compound D3 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl isobutyrate;

[0027] Compound D4 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pivalate;

[0028] Compound D5 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzoate;

[0029] Compound D6 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethyl)benzoate;

[0030] Compound D7 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzoate;

[0031] Compound D8 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-fluorobenzoate;

[0032] Compound D9 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl cyclopropanecarboxylate;

[0033] Compound D10 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzoate;

[0034] Compound D11 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-methylbenzoate;

[0035] Compound D12 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-fluorobenzoate;

[0036] Compound D13 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-chlorobenzoate;

[0037] Compound D14 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-chlorobenzoate;

[0038] Compound D15 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-bromobenzoate;

[0039] Compound D16 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzoate;

[0040] Compound D17 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-nitrobenzenesulfonate;

[0041] Compound D18 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-fluorobenzenesulfonate;

[0042] Compound D19 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzenesulfonate;

[0043] Compound D20 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzenesulfonate;

[0044] Compound D21 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzenesulfonate;

[0045] Compound D22 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzenesulfonate;

[0046] Compound D23 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-(trifluoromethyl)benzoate;

[0047] Compound E1 is (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-thione;

[0048] Compound E2 is 1-methyl-9-(3,4,5-trimethoxybenzyl)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one;

[0049] Compound E3 is (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one;

[0050] Compound F1 is (E)-1,5-dimethyl-6-(3,4,5-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0051] Compound F2 is (E)-1,5-dimethyl-6-(2,4,6-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0052] Compound F3 is (E)-6-(4-hydroxy-3,5-dimethoxystyryl)-1,5-dimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one.

[0053] Use of compounds D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D20, D21, D22, D23, E1, E2, E3, F1, F2, F3 in the tricyclic pyrazolopyrimidine aryl ester derivatives for the preparation of drugs for treating HCT116 colon cancer.

[0054] Use of compounds D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D20, D21, D22, D23, E1, E2, E3, F1, F2, F3 in the tricyclic pyrazolopyrimidine aryl ester derivatives for the preparation of drugs for treating HGC27 gastric cancer.

[0055] The preparation method of the tricyclic pyrazolopyrimidine aryl ester derivatives described in the present invention is carried out according to the following steps:

[0056] Preparation of compound A:

[0057] Dissolve 20 g (118.2 mmol) of ethyl 5-amino-1-methylpyrazole-4-carboxylate in 150 mL of anhydrous 1,2-dichloroethane, add 23.4 g (236.4 mmol) of 2-azepanone, and then slowly dropwise add 45.32 g (295.6 mmol) of phosphorus oxychloride. React under reflux at 80 °C for 8 hours. Detect by TLC until the raw materials react completely. Pour the reaction solution into 200 mL of water to quench, adjust the pH to 7 with an aqueous NaOH solution, then add ethyl acetate (50 mL × 3) for liquid-liquid extraction. Combine the organic phases, wash with 100 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate. The dried product is slurried with 50 mL of tetrahydrofuran to obtain 22.1 g of white solid compound A, 1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one, with a yield of 91.7%.

[0058] Preparation of compound B:

[0059] 10 g (49.0 mmol) of 1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one (Compound A) was dissolved in 150 mL of methanol. 3.92 g (98.0 mmol) of sodium hydroxide and 14.41 g (73.4 mmol) of 3,4,5-trimethoxybenzaldehyde were added. The mixture was refluxed at 90 °C for 8 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was directly filtered, and the filter cake was washed with 30 mL of methanol and 30 mL of ethyl acetate respectively to obtain 15.1 g of pale yellow solid Compound B, (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one, with a yield of 80.1%.

[0060] Preparation of Compound C:

[0061] 10 g (26.1 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one (Compound B) was dissolved in 100 mL of acetic acid solution of 40% hydrobromic acid. The mixture was refluxed at 90 °C for 1 hour. The reaction was monitored by TLC and found to be complete. 100 mL of water was added to the reaction solution to quench the reaction. After solid precipitation, the mixture was filtered, and the filter cake was washed twice with 30 mL of methanol to obtain 8.2 g of yellow solid Compound C, (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one, with a yield of 85.5%.

[0062] Preparation of Compounds D1 - D23:

[0063] 0.54 mmol of the obtained (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one (Compound C) was dissolved in anhydrous dichloromethane. Then, 1.30 mmol of the acid-binding agent triethylamine and 0.27 mmol of the catalyst 4-dimethylaminopyridine were added dropwise. 0.81 mmol of the acyl chloride diluted with dichloromethane was slowly added dropwise at 0 °C. The mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC and completed. Water was added to remove the excess acyl chloride. The mixture was extracted with dichloromethane (5 mL × 3), washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the organic phases were combined, concentrated, and purified by silica gel column chromatography. Elution was carried out with a petroleum ether:ethyl acetate eluent with a volume ratio of 1:1 to obtain the target compounds D1 - D23.

[0064] Preparation of Compound E1:

[0065] Dissolve 200 mg (0.52 mmol) of (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one (Compound B) in 5 mL of toluene solution. Add 317.30 mg (0.78 mmol) of Lawesson's reagent and reflux the reaction at 90 °C for 8 hours. Monitor the reaction completion by TLC. Add 10 mL of water to the reaction solution and extract with ethyl acetate (10 mL × 3). Combine the organic phases and wash with 15 mL of saturated brine. Concentrate and dry, and purify by silica gel column chromatography using a petroleum ether and ethyl acetate eluent with a volume ratio of 3:1 to obtain 157.0 mg of yellow solid Compound E with a yield of 75.6%.

[0066] Preparation of Compound E2:

[0067] Dissolve 200 mg (0.98 mmol) of (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one (Compound B) in 5 mL of methanol solution. Add 50 mg of palladium on carbon and react at room temperature for 8 hours. Monitor the reaction completion by TLC. Filter the reaction solution directly to remove the palladium on carbon, concentrate the reaction solution to obtain 180.0 mg of yellow solid Compound E2 with a yield of 90%.

[0068] Preparation of Compound E3:

[0069] Dissolve 10 g (26.1 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one (Compound B) in 100 mL of acetic acid solution containing 40% hydrobromic acid. Reflux the reaction at 90 °C for 1 hour. Monitor the reaction completion by TLC. Quench the reaction solution by adding 100 mL of water. Filter after solid precipitation. Wash the filter cake twice with 30 mL of methanol each to obtain 8.2 g of yellow solid Compound E3, which is (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-4(1H)-one with a yield of 85.5%.

[0070] Preparation of Compound F1:

[0071] Step 1: Dissolve 5 g (29.55 mmol) of ethyl 5-amino-1-methylpyrazole-4-carboxylate in 70 mL of anhydrous 1,2-dichloroethane. Add 3.24 g (44.33 mmol) of N-methylacetamide, and then slowly dropwise add 11.33 g (73.89 mmol) of phosphorus oxychloride. Reflux the reaction mixture at 80 °C for 8 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 100 mL of water to quench the reaction. Adjust the pH to 7 with an aqueous NaOH solution, then extract with ethyl acetate (30 mL × 3). Combine the organic phases, wash with 100 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate, and slurry the dried product in 50 mL of tetrahydrofuran to obtain 4.8 g of white solid compound 1,5,6-trimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (Compound F0), with a yield of 91.1%;

[0072] Step 2: Dissolve 200 mg (1.12 mmol) of 1,5,6-trimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (Compound F0) in 5 mL of methanol. Add 89.79 mg (2.24 mmol) of sodium hydroxide and 330.30 mg (1.68 mmol) of 3,4,5-trimethoxybenzaldehyde. Reflux the reaction mixture at 90 °C for 8 hours. Monitor the reaction by TLC until it is complete. Filter the reaction solution directly, and wash the filter cake with 10 mL of methanol and 10 mL of ethyl acetate respectively to obtain 180 mg of light green solid compound F1 (E)-1,5-dimethyl-6-(3,4,5-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, with a yield of 45.1%;

[0073] Preparation of Compound F2:

[0074] Dissolve 200 mg (1.12 mmol) of 1,5,6-trimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (Compound F0) in 5 mL of methanol. Add 89.79 mg (2.24 mmol) of sodium hydroxide and 330.30 mg (1.68 mmol) of 2,4,6-trimethoxybenzaldehyde. Reflux the reaction mixture at 90 °C for 8 hours. Monitor the reaction by TLC until it is complete. Filter the reaction solution directly, and wash the filter cake with 10 mL of methanol and 10 mL of ethyl acetate respectively to obtain 175 mg of light green solid compound F2 (E)-1,5-dimethyl-6-(2,4,6-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, with a yield of 43.75%;

[0075] Preparation of Compound F3:

[0076] Dissolve 50 mg (0.140 mmol) of compound F1 (E)-1,5-dimethyl-6-(3,4,5-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one in 5 mL of an acetic acid solution of 40% hydrobromic acid, reflux the reaction at 90 °C for 1 hour. When the reaction is complete as detected by TLC, add 100 mL of water to the reaction solution. After solid precipitation occurs, filter, and wash the filter cake twice with 3 mL of methanol each time to obtain 21 mg of yellow solid compound F3 (E)-6-(4-hydroxy-3,5-dimethoxystyryl)-1,5-dimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, with a yield of 39.1%.

[0077] A tricyclic pyrazolopyrimidine aryl ester derivative described in the present invention uses ethyl 5-amino-1-methylpyrazole-4-carboxylate as the starting material, and through three-step reactions, a monosubstituted (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (C) is obtained. Then, under the action of a catalytic amount of base, it reacts with acyl chlorides with different substitutions to obtain the target (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one derivatives (D1-D23). The synthetic route is as follows:

[0078]

[0079] The results of in vitro activity screening of this class of compounds show that: using doxorubicin (DOX) as the positive control drug, 29 compounds all have good anti-cancer activity (IC 50 <5 μM), among which 24 compounds show excellent activity in inhibiting HCT116 colon cancer cells (IC 50 <1 μM), especially D2, D3, D4, D9, D11, D14, and E3 have the best activity (IC 50 = 0.01 μM). Among them, 26 compounds show excellent activity in inhibiting HGC27 gastric cancer cells (IC 50 <1 μM), especially D3, D7, D8, D9, D15, and E3 have the best activity (IC 50 equal to or less than 0.05 μM). Detailed implementation methods

[0080] The present invention is further illustrated based on the examples, but the present invention is not limited to these examples;

[0081] Reagents: All reagents are commercially available analytical pure.

[0082] Example 1

[0083] Preparation of diethyl 2,5-diamino-3,4-thiophenedicarboxylate (A):

[0084] Dissolve 20 g (118.2 mmol) of ethyl 5-amino-1-methylpyrazole-4-carboxylate in 150 mL of anhydrous 1,2-dichloroethane, add 23.4 g (236.4 mmol) of 2-azepanone, and then slowly dropwise add 45.32 g (295.6 mmol) of phosphorus oxychloride. Reflux the reaction mixture at 80 °C for 8 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 200 mL of water to quench the reaction, adjust the pH to 7 with NaOH, then extract with ethyl acetate (50 mL × 3). Wash with 100 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent. Pulverize the dried product with 50 mL of tetrahydrofuran to obtain 2.1 g of white solid compound A, 1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one, with a yield of 91.7%, melting point: 154 - 155 °C;

[0085] 1 H NMR (400 MHz, CDCl 3 ) δ 5.25 (s, 4H), 4.26–4.21 (m, 4H), 1.30 (t, J = 7.1 Hz, 6H). HRMS (ESI) m / z: 205.2410.

[0086] Example 2

[0087] Preparation of (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (B):

[0088] Dissolve 10 g (49.0 mmol) of 1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (compound A) in 150 mL of methanol, add 3.92 g (98.0 mmol) of sodium hydroxide and 14.41 g (73.4 mmol) of 3,4,5-trimethoxybenzaldehyde. Reflux the reaction mixture at 90 °C for 8 hours. Monitor the reaction by TLC until it is complete. Filter the reaction solution directly, and wash the filter cake with 30 mL of methanol and 30 mL of ethyl acetate respectively to obtain 15.1 g of light yellow solid compound B, (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one, with a yield of 80.1%; melting point: 254 - 255 °C;

[0089] 1 H NMR (500 MHz, DMSO) δ 8.10 (s, 1H), 8.04 (s, 1H), 6.85 (s, 2H), 4.02–3.97 (m, 2H), 3.95 (s, 3H), 3.82 (s, 6H), 3.70 (s, 3H), 3.15 (d, J = 3.7 Hz, 2H), 2.94 (td, J = 6.6, 2.2 Hz, 2H), 1.91 (qd, J = 6.3, 3.4 Hz, 2H). HRMS (ESI) m / z: 383.1706。

[0090] Example 3

[0091] Preparation of compound (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (C):

[0092] Dissolve 10 g (26.1 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound B) in 100 mL of a 40% hydrobromic acid acetic acid solution, reflux at 90 °C for 1 hour. When the reaction is complete by TLC detection, add 100 mL of water to the reaction solution. After solid precipitation, filter. Wash the filter cake twice with 30 mL of methanol each to obtain 8.2 g of yellow solid compound C (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one, with a yield of 85.5%;

[0093] 1 H NMR (500 MHz, DMSO) δ 8.89 (s, 1H), 8.07 (d, J = 2.2 Hz, 1H), 8.02 (s, 1H), 6.85 (s, 2H), 4.01–3.97 (m, 2H), 3.94 (s, 3H), 3.81 (s, 5H), 2.98–2.91 (m, 2H), 1.92 (p, J = 6.3 Hz, 2H). HRMS (ESI) m / z: 369.1553。

[0094] Example 4

[0095] Preparation of (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pentanoate (D1):

[0096] Dissolve 50 mg (0.14 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 5 mL of anhydrous dichloromethane. Add 24.6 mg (0.20 mmol) of valeryl chloride at 0 °C, then successively add 33 mg (0.32 mmol) of triethylamine and 8.3 mg (0.07 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (5 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent. Purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 20 mg of the target compound D1 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl valerate, with a yield of 32.2%, melting point: 202 - 203 °C;

[0097] 1 H NMR (500 MHz, CDCl 3 ) δ 8.16 (d, J = 2.1 Hz, 1H), 8.06 (s, 1H), 6.69 (s, 2H), 4.22–4.07 (m, 2H), 4.03 (s, 3H), 3.85 (s, 6H), 2.95 (td, J = 6.6, 2.1 Hz, 2H), 2.63 (t, J = 7.5 Hz, 2H), 2.08–1.93 (m, 2H), 1.78 (p, J = 7.5 Hz, 2H), 1.54–1.42 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H). HRMS (ESI) m / z: 453.2111.

[0098] Example 5

[0099] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl dodecanoate (D2):

[0100] Dissolve 50 mg (0.14 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 5 mL of anhydrous dichloromethane. At 0 °C, add 44.5 mg (0.20 mmol) of dodecanoyl chloride, then successively add 33 mg (0.32 mmol) of triethylamine and 8.3 mg (0.07 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (5 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 31 mg of the target compound D2 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl dodecanoate, with a yield of 41.5%, melting point: 212 - 213 °C;

[0101] 1 H NMR(600MHz,CDCl 3 )δ8.16(d,J=2.1Hz,1H),8.06(s,1H),6.69(s,2H),4.16–4.09(m,2H),4.03(s,3H),3.85(s,6H),2.95(td,J=6.5,2.1Hz,2H),2.63(t,J=7.4Hz,2H),2.01(p,J=6.3Hz,2H),1.78(p,J=7.5Hz,2H),1.47–1.41(m,2H),1.27(q,J=10.8Hz,14H),0.88(t,J=7.0Hz,3H).HRMS(ESI)m / z:551.3224。

[0102] Example 6

[0103] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl isobutyrate (D3):

[0104] Dissolve 100 mg (0.28 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. Add 43.38 mg (0.40 mmol) of isobutyryl chloride at 0 °C, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 62 mg of the target compound D3 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl isobutyrate, with a yield of 52.1%, melting point: 207 - 208 °C;

[0105] 1 H NMR(500MHz,CDCl 3 )δ8.16(d,J=2.1Hz,1H),8.07(s,1H),6.69(s,2H),4.15–4.11(m,2H),4.03(s,3H),3.85(s,6H),2.95(td,J=6.6,2.1Hz,2H),2.93–2.87(m,1H),2.03–1.99(m,2H),1.35(d,J=7.0Hz,6H).HRMS(ESI)m / z:439.1958。

[0106] Example 7

[0107] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pivalate (D4):

[0108] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 98.19 mg (0.81 mmol) of pivaloyl chloride, then successively add 131.85 mg (1.30 mmol) of triethylamine and 33.16 mg (0.27 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent. Purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 121 mg of the target compound D4 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pivalate, with a yield of 49.26%, melting point: 278 - 279 °C;

[0109] 1 H NMR (500 MHz, MeOD) δ 8.26 (s, 1H), 8.02 (s, 1H), 6.90–6.79 (m, 2H), 4.15–4.10 (m, 2H), 4.02 (s, 3H), 3.85 (s, 6H), 3.02 (td, J = 6.3, 2.2 Hz, 2H), 2.08–1.96 (m, 2H), 1.37 (s, 9H). HRMS (ESI) m / z: 453.2111.

[0110] Example 8

[0111] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzoate (D5):

[0112] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. Add 178.72 mg (0.81 mmol) of 4-bromobenzoyl chloride at 0 °C, then successively add 131.85 mg (1.30 mmol) of triethylamine and 33.16 mg (0.27 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 150 mg of the target compound D5 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzoate, with a yield of 50.1%, melting point: 262 - 263 °C;

[0113] 1 H NMR (500 MHz, DMSO) δ 8.18 (d, J = 2.2 Hz, 1H), 8.08–8.05 (m, 1H), 8.05–8.03 (m, 1H), 7.90 (dd, J = 8.6, 1.7 Hz, 1H), 7.85–7.82 (m, 1H), 7.73 (t, J = 8.7 Hz, 1H), 6.98 (s, 2H), 4.04–4.01 (m, 2H), 3.98 (d, J = 1.2 Hz, 3H), 3.82 (s, 6H), 3.02–2.93 (m, 2H), 1.98–1.92 (m, 2H). HRMS (ESI) m / z: 553.0892.

[0114] Example 9

[0115] Preparation of (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethyl)benzoate (D6):

[0116] Dissolve 100 mg (0.28 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. Add 169.84 mg (0.81 mmol) of 4-(trifluoromethyl)benzoyl chloride at 0 °C, then successively add 131.85 mg (1.30 mmol) and 33.16 mg (0.27 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 172 mg of the target compound D6 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethyl)benzoate, with a yield of 58.6%, melting point: 257 - 258 °C;

[0117] 1 H NMR (600 MHz, DMSO) δ 8.33 (s, 2H), 8.18 (s, 1H), 8.06 (s, 1H), 7.99 (d, J = 8.4 Hz, 2H), 6.98 (s, 2H), 4.02 (dd, J = 6.7, 4.9 Hz, 2H), 3.97 (s, 3H), 3.82 (s, 6H), 3.02–2.95 (m, 2H), 1.95 (p, J = 6.4 Hz, 2H). HRMS (ESI) m / z: 541.1689.

[0118] Example 10

[0119] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzoate (D7):

[0120] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 182.87 mg (0.81 mmol) of 4-(trifluoromethoxy)benzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. When TLC shows that the raw materials have completely reacted, pour the reaction solution into 10 mL of water to quench the reaction, then add ethyl acetate (10 mL × 3) for liquid-liquid extraction, wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 62 mg of the target compound D7 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzoate, with a yield of 52.1%, melting point: 207 - 208 °C;

[0121] 1 H NMR(500MHz,CDCl 3 )δ8.36–8.25(m,2H),8.19(d,J=2.2Hz,1H),8.06(s,1H),7.34(dq,J=7.9,1.1Hz,2H),6.74(s,2H),4.17–4.10(m,2H),4.03(s,3H),3.85(s,6H),2.97(td,J=6.5,2.1Hz,2H),2.03(dtd,J=7.7,6.0,3.7Hz,2H).HRMS(ESI)m / z:557.1638。

[0122] Example 11

[0123] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-fluorobenzoate (D8):

[0124] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 129.12 mg (0.81 mmol) of 2-fluorobenzoyl chloride, then successively add 131.85 mg (1.30 mmol) of triethylamine and 33.16 mg (0.27 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 178 mg of the target compound D8 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-fluorobenzoate (D8), with a yield of 66.8%, melting point: 264 - 265 °C;

[0125] 1 H NMR (500 MHz, DMSO) δ 8.17 (d, J = 2.2 Hz, 1H), 8.08 (dd, J = 7.6, 1.8 Hz, 1H), 8.06 (s, 1H), 7.82–7.75 (m, 1H), 7.48–7.38 (m, 2H), 6.96 (s, 2H), 4.04–3.99 (m, 2H), 3.96 (s, 3H), 3.82 (s, 6H), 3.09–2.89 (m, 2H), 1.94 (t, J = 5.9 Hz, 2H). HRMS (ESI) m / z: 491.1711.

[0126] Example 12

[0127] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl cyclopropanecarboxylate (D9):

[0128] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 85.12 mg (0.81 mmol) of cyclopropylcarbonyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 106 mg of the target compound D9 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl cyclopropanecarboxylate, with a yield of 44.7%, melting point: 237 - 238 °C;

[0129] 1 H NMR (600 MHz, DMSO) δ 8.13 (d, J = 2.3 Hz, 1H), 8.06 (s, 1H), 6.89 (s, 2H), 4.00 (dd, J = 7.2, 4.5 Hz, 2H), 3.96 (s, 3H), 3.80 (s, 6H), 2.99–2.90 (m, 2H), 1.92 (t, J = 6.0 Hz, 2H), 1.91–1.88 (m, 1H), 1.07–1.03 (m, 2H), 0.99–0.95 (m, 2H). HRMS (ESI) m / z: 437.1803.

[0130] Example 13

[0131] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzoate (D10):

[0132] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 134.84 mg (0.81 mmol) of 4-cyanobenzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 123 mg of the target compound D10 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzoate, with a yield of 45.5%, melting point: 257 - 258 °C;

[0133] 1 H NMR(500MHz,CDCl 3 )δ8.38–8.30(m,1H),8.23–8.14(m,1H),7.86–7.78(m,1H),6.74(s,1H),4.16–4.12(m,1H),4.04(s,2H),3.85(s,3H),3.00–2.94(m,1H),2.06–2.01(m,1H).HRMS(ESI)m / z:498.1758。

[0134] Example 14

[0135] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-methylbenzoate (D11):

[0136] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 125.89 mg (0.81 mmol) of 4-methylbenzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 162 mg of the target compound D11 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-methylbenzoate, with a yield of 61.3%, melting point: 237 - 238 °C;

[0137] 1 H NMR (600 MHz, DMSO) δ 8.18 (d, J = 2.3 Hz, 1H), 8.07 (s, 1H), 8.00 (d, J = 8.1 Hz, 2H), 7.41 (d, J = 7.9 Hz, 2H), 6.96 (s, 2H), 4.03–4.00 (m, 2H), 3.97 (s, 3H), 3.80 (s, 6H), 3.00–2.97 (m, 2H), 2.42 (s, 3H), 1.95 (t, J = 5.9 Hz, 2H). HRMS (ESI) m / z: 487.1961.

[0138] Example 15

[0139] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-fluorobenzoate (D12):

[0140] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 129.12 mg (0.81 mmol) of 3-fluorobenzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 153 mg of the target compound D12 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-fluorobenzoate, with a yield of 53.5%, melting point: 294 - 295 °C;

[0141] 1 H NMR (600 MHz, DMSO) δ 8.18 (s, 1H), 8.07 (s, 1H), 7.99–7.96 (m, 2H), 7.85 (dd, J = 9.1, 2.5 Hz, 2H), 6.97 (s, 2H), 4.01 (d, J = 5.8 Hz, 2H), 3.97 (s, 3H), 3.82 (s, 6H), 2.98 (d, J = 6.5 Hz, 2H), 1.95 (t, J = 5.9 Hz, 2H). HRMS (ESI) m / z: 491.1709.

[0142] Example 16

[0143] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-chlorobenzoate (D13):

[0144] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 43.38 mg (0.40 mmol) of isobutyryl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 62 mg of the target compound D13 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-chlorobenzoate, with a yield of 52.1% and a melting point of 207 - 208 °C;

[0145] 1 H NMR (600 MHz, DMSO) δ 8.18 (d, J = 2.2 Hz, 1H), 8.13–8.11 (m, 2H), 8.07 (s, 1H), 7.71–7.67 (m, 2H), 6.97 (s, 2H), 4.02 (dd, J = 7.1, 4.6 Hz, 2H), 3.97 (s, 3H), 3.81 (s, 6H), 3.02–2.96 (m, 3H), 1.95 (q, J = 6.1 Hz, 2H). HRMS (ESI) m / z: 507.1433.

[0146] Example 17

[0147] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-chlorobenzoate (D14):

[0148] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 142.52 mg (0.81 mmol) of 3-chlorobenzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent. Purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 143 mg of the target compound D14 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-chlorobenzoate, with a yield of 51.6%, melting point: 276 - 277 °C;

[0149] 1 H NMR (600 MHz, DMSO) δ 8.17 (d, J = 2.3 Hz, 1H), 8.08 (dt, J = 7.3, 1.5 Hz, 2H), 8.06 (s, 1H), 7.88–7.83 (m, 1H), 7.68–7.63 (m, 1H), 6.97 (s, 2H), 4.03–4.00 (m, 2H), 3.97 (s, 3H), 3.82 (s, 6H), 3.02–2.93 (m, 2H), 1.95 (h, J = 5.2 Hz, 2H). HRMS (ESI) m / z: 507.1428.

[0150] Example 18

[0151] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-bromobenzoate (D15):

[0152] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 178.71 mg (0.81 mmol) of 3-bromobenzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 164 mg of the target compound D15 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-bromobenzoate, with a yield of 54.7%, melting point: 257 - 258 °C;

[0153] 1 H NMR(500MHz,CDCl 3 )δ8.37(t,J=1.8Hz,1H),8.21–8.11(m,2H),8.06(s,1H),7.76(ddd,J=8.0,2.1,1.1Hz,1H),7.40(d,J=7.9Hz,1H),6.73(s,2H),4.15–4.12(m,2H),4.03(s,3H),3.84(s,6H),2.97(td,J=6.6,2.2Hz,2H),2.03–1.98(m,2H).HRMS(ESI)m / z:553.0882。

[0154] Example 19

[0155] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzoate (D16):

[0156] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 160.16 mg (0.81 mmol) of 4-(tert-butyl)benzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 145 mg of the target compound D16 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzoate, with a yield of 50.5%, melting point: 254 - 255 °C;

[0157] 1 H NMR(500MHz,CDCl 3 )δ8.20–8.17(m,2H),8.17(d,J=1.9Hz,1H),8.06(d,J=1.6Hz,1H),7.56–7.49(m,2H),6.73(s,2H),4.16–4.11(m,2H),4.03(s,3H),3.83(s,6H),2.97(td,J=6.6,2.1Hz,2H),2.06–2.00(m,2H),1.37(s,9H).HRMS(ESI)m / z:529.2443。

[0158] Example 20

[0159] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-nitrobenzenesulfonate (D17):

[0160] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 180.47 mg (0.81 mmol) of 4-nitrobenzenesulfonyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. When TLC shows that the raw materials have completely reacted, pour the reaction solution into 10 mL of water to quench the reaction, then add ethyl acetate (10 mL × 3) for liquid-liquid extraction, wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 166 mg of the target compound D17 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-nitrobenzenesulfonate, with a yield of 66.6%, melting point: 281 - 282 °C;

[0161] 1 H NMR (500 MHz, DMSO) δ 8.48 (d, J = 8.7 Hz, 2H), 8.26–8.15 (m, 2H), 8.10 (d, J = 2.4 Hz, 1H), 8.06 (s, 1H), 6.88 (s, 2H), 3.99 (t, J = 5.7 Hz, 2H), 3.95 (s, 3H), 3.63 (s, 6H), 3.01–2.85 (m, 2H), 1.92 (dq, J = 11.9, 6.1 Hz, 2H). HRMS (ESI) m / z: 554.1336.

[0162] Example 21

[0163] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-fluorobenzenesulfonate (D18):

[0164] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 158.48 mg (0.81 mmol) of 4-fluorobenzenesulfonyl chloride, then sequentially add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. When TLC shows that the raw materials have completely reacted, pour the reaction solution into 10 mL of water to quench the reaction, then add ethyl acetate (10 mL × 3) for liquid-liquid extraction, wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 144 mg of the target compound D18 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-fluorobenzenesulfonate, with a yield of 50.4%, melting point: 274 - 275 °C;

[0165] 1 H NMR (600 MHz, DMSO) δ 8.10 (d, J = 2.2 Hz, 1H), 8.06 (s, 1H), 8.00–7.95 (m, 2H), 7.55–7.51 (m, 2H), 6.87 (s, 2H), 3.99 (dd, J = 6.8, 4.7 Hz, 2H), 3.95 (d, J = 2.9 Hz, 3H), 3.65 (s, 6H), 2.95–2.90 (m, 2H), 1.92 (p, J = 6.4 Hz, 2H). HRMS (ESI) m / z: 527.1389.

[0166] Example 22

[0167] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzenesulfonate (D19):

[0168] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 212.24 mg (0.81 mmol) of 4-(trifluoromethoxy)benzenesulfonyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 159 mg of the target compound D19, (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzenesulfonate, with a yield of 49.4%, melting point: 286 - 287 °C;

[0169] 1 H NMR (600 MHz, DMSO) δ 8.10 (d, J = 2.3 Hz, 1H), 8.06 (s, 1H), 8.05–8.01 (m, 2H), 7.68 (d, J = 8.5 Hz, 2H), 6.87 (s, 2H), 4.02–3.97 (m, 2H), 3.95 (s, 3H), 3.62 (s, 6H), 2.95–2.90 (m, 2H), 1.92 (dt, J = 6.3, 3.5 Hz, 2H). HRMS (ESI) m / z: 593.1313.

[0170] Example 23

[0171] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzenesulfonate (D20):

[0172] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. Add 208.08 mg (0.81 mmol) of 4-bromobenzenesulfonyl chloride at 0 °C, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 162 mg of the target compound D20 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzenesulfonate, with a yield of 50.8%, melting point: 288 - 289 °C;

[0173] 1 H NMR (600 MHz, DMSO) δ 8.10 (s, 1H), 8.06 (s, 1H), 7.95–7.89 (m, 2H), 7.82 (d, J = 8.6 Hz, 2H), 6.87 (s, 2H), 4.00 (t, J = 5.7 Hz, 2H), 3.95 (s, 2H), 3.64 (s, 6H), 2.93 (t, J = 6.2 Hz, 2H), 1.92 (d, J = 6.1 Hz, 3H). HRMS (ESI) m / z: 589.0568.

[0174] Example 24

[0175] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzenesulfonate (D21):

[0176] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. Add 164.19 mg (0.81 mmol) of 4-cyanobenzenesulfonyl chloride at 0 °C, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. When TLC shows that the raw materials have completely reacted, pour the reaction solution into 10 mL of water to quench the reaction. Then add ethyl acetate (10 mL × 3) for liquid-liquid extraction, wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 115 mg of the target compound D21 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzenesulfonate, with a yield of 39.7%, melting point: 250 - 251 °C;

[0177] 1 H NMR (600 MHz, DMSO) δ 8.18–8.16 (m, 2H), 8.10 (d, J = 2.3 Hz, 1H), 8.09 (d, J = 1.9 Hz, 1H), 8.08 (d, J = 1.8 Hz, 1H), 8.06 (s, 1H), 6.88 (s, 2H), 4.00 (t, J = 5.8 Hz, 2H), 3.95 (s, 3H), 3.63 (s, 6H), 2.95–2.91 (m, 2H), 1.93–1.91 (m, 2H). HRMS (ESI) m / z: 534.1447.

[0178] Example 25

[0179] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzenesulfonate (D22):

[0180] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 189.52 mg (0.81 mmol) of 4-(tert-butyl)benzenesulfonyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 10 mL of water to quench the reaction, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 155 mg of the target compound D22 (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzenesulfonate, with a yield of 50.6%, melting point: 248 - 249 °C;

[0181] 1 H NMR(500MHz,CDCl 3 )δ8.13(d,J=2.2Hz,1H),8.06(s,1H),7.96–7.90(m,2H),7.58–7.55(m,2H),6.62(s,2H),4.15–4.12(m,2H),4.02(s,3H),3.70(s,6H),2.92(td,J=6.6,2.1Hz,2H),2.01(q,J=6.0Hz,2H),1.37(s,9H).HRMS(ESI)m / z:565.2124。

[0182] Example 26

[0183] Preparation of compound (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-(trifluoromethyl)benzoate (D23):

[0184] Dissolve 200 mg (0.56 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound C) in 10 mL of anhydrous dichloromethane. At 0 °C, add 168.84 mg (0.81 mmol) of 2-(trifluoromethyl)benzoyl chloride, then successively add 66 mg (0.64 mmol) of triethylamine and 16.6 mg (0.14 mmol) of 4-dimethylaminopyridine. React at room temperature for 3 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Quench the reaction mixture by pouring it into 10 mL of water, then extract with ethyl acetate (10 mL × 3) by liquid separation. Wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate the solvent, and purify the dried product by Prep-TLC with a mobile phase polarity of petroleum ether:ethyl acetate (volume ratio 1:3) to obtain 145 mg of the target compound D, (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-(trifluoromethyl)benzoate, with a yield of 49.4%, melting point: 266 - 267 °C;

[0185] 1 H NMR (600 MHz, DMSO) δ 8.17 (d, J = 2.3 Hz, 1H), 8.13–8.09 (m, 1H), 8.07 (s, 1H), 8.00–7.97 (m, 1H), 7.92–7.87 (m, 2H), 6.98 (s, 2H), 4.03–4.00 (m, 2H), 3.97 (s, 3H), 3.85 (s, 6H), 3.00–2.96 (m, 2H), 1.97–1.92 (m, 2H). HRMS (ESI) m / z: 541.1683.

[0186] Example 27

[0187] Preparation of compound (E)-1-methyl-9-(3,4,5-trimethoxybenzylidene)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidine-4(1H)-thione (E1):

[0188] Dissolve 200 mg (0.52 mmol) of (E)-1-methyl-9-(3,4,5-trimethoxybenzyl)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound B) in 5 mL of toluene solution. Add 317.30 mg (0.78 mmol) of Lawesson's reagent and reflux the reaction at 90 °C for 8 hours. Monitor the reaction by TLC until completion. Add 10 mL of water to the reaction solution, extract the organic phase with ethyl acetate (10 mL × 2), wash it with 15 mL of saturated brine, combine the organic phases, concentrate and dry, and purify by silica gel column chromatography using a petroleum ether and ethyl acetate eluent with a volume ratio of 3:1 to obtain 157.0 mg of yellow solid Compound E1 (E)-1-methyl-9-(3,4,5-trimethoxybenzyl)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-thione, with a yield of 75.6%, melting point: 211 - 212 °C;

[0189] 1 H NMR(500MHz,CDCl 3 )δ8.23(s,1H),8.10(t,J=2.2Hz,1H),6.70(s,2H),4.69–4.63(m,2H),4.03(s,3H),3.91(t,J=0.8Hz,9H),2.99(td,J=6.6,2.2Hz,2H),2.13–2.04(m,2H).HRMS(ESI)m / z:399.1477。

[0190] Example 28

[0191] Preparation of compound 1-methyl-9-(3,4,5-trimethoxybenzyl)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (E2):

[0192] Dissolve 200 mg (0.98 mmol) of (E)-1-methyl-9-(3,4,5-trimethoxybenzyl)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound B) in 5 mL of methanol solution. Add 50 mg of palladium on carbon and react at room temperature for 8 hours. Monitor the reaction by TLC until completion. Filter the reaction solution directly to remove the palladium on carbon, concentrate the reaction solution to obtain 180.0 mg of yellow solid compound E2 1-methyl-9-(3,4,5-trimethoxybenzyl)-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one, with a yield of 90%, melting point: 277 - 278 °C;

[0193] 1 1H NMR (600 MHz, DMSO) δ 8.03 (s, 1H), 6.57 (s, 2H), 4.18 (dt, J = 13.6, 5.4 Hz, 1H), 3.90 (s, 3H), 3.73 (s, 6H), 3.71–3.67 (m, 1H), 3.62 (s, 3H), 3.45 (dd, J = 13.6, 4.7 Hz, 1H), 3.29–3.22 (m, 1H), 2.74 (dd, J = 13.7, 9.4 Hz, 1H), 1.89–1.78 (m, 3H), 1.47–1.41 (m, 1H). HRMS (ESI) m / z: 385.1859。

[0194] Example 29

[0195] Preparation of compound (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (E3):

[0196] 10 g (26.1 mmol) of (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one (Compound B) was dissolved in 100 mL of an acetic acid solution of 40% hydrobromic acid. The mixture was refluxed at 90 °C for 1 hour. The reaction was monitored by TLC and was found to be complete. 100 mL of water was added to the reaction mixture to quench the reaction. A solid precipitate formed and was filtered. The filter cake was washed twice with 30 mL of methanol each time to obtain 8.2 g of the yellow solid compound E3, (E)-9-(4-hydroxy-3,5-dimethoxybenzylidene)-1-methyl-6,7,8,9-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-4(1H)-one, with a yield of 85.5%.

[0197] Example 30

[0198] Preparation of compound (E)-1,5-dimethyl-6-(3,4,5-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (F1):

[0199] Step 1: Dissolve 5 g (29.55 mmol) of ethyl 5-amino-1-methylpyrazole-4-carboxylate in 70 mL of anhydrous 1,2-dichloroethane. Add 3.24 g (44.33 mmol) of N-methylacetamide, and then slowly dropwise add 11.33 g (73.89 mmol) of phosphorus oxychloride. Reflux the reaction at 80 °C for 8 hours. Monitor the reaction by TLC until the raw materials are completely reacted. Pour the reaction solution into 100 mL of water to quench the reaction. Adjust the pH to 7 with an aqueous solution of NaOH, then add ethyl acetate (30 mL × 3) for liquid-liquid extraction. Combine the organic phases, wash with 100 mL of saturated brine, dry over anhydrous sodium sulfate, concentrate, and slurry the dried product in 50 mL of tetrahydrofuran to obtain 4.8 g of the white solid compound 1,5,6-trimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (Compound F0), with a yield of 91.1%;

[0200] Step 2: Dissolve 200 mg (1.12 mmol) of Compound F0 in 5 mL of methanol. Add 89.79 mg (2.24 mmol) of sodium hydroxide and 330.30 mg (1.68 mmol) of 3,4,5-trimethoxybenzaldehyde. Reflux the reaction at 90 °C for 8 hours. Monitor the reaction by TLC until it is complete. Filter the reaction solution directly, and wash the filter cake with 10 mL of methanol and 10 mL of ethyl acetate respectively to obtain 180 mg of the light green solid compound (E)-1,5-dimethyl-6-(3,4,5-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (Compound F1), with a yield of 45.1% and a melting point of 189 - 190 °C;

[0201] 1 H NMR (400 MHz, CDCl 3 ) δ 8.33–8.04 (m, 1H), 7.92 (d, J = 15.2 Hz, 1H), 6.98 (dd, J = 15.2, 4.9 Hz, 1H), 6.88 (d, J = 49.2 Hz, 2H), 4.02 (d, J = 0.8 Hz, 3H), 3.94 (s, 3H), 3.92–3.89 (m, 6H), 3.71 (d, J = 6.4 Hz, 3H). HRMS (ESI) m / z: 357.1552.

[0202] Example 31

[0203] Preparation of compound (E)-1,5-dimethyl-6-(2,4,6-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (F2):

[0204] Dissolve 200 mg (1.12 mmol) of compound F0 in 5 mL of methanol, add 89.79 mg (2.24 mmol) of sodium hydroxide and 330.30 mg (1.68 mmol) of 2,4,6-trimethoxybenzaldehyde, and reflux the reaction mixture at 90 °C for 8 hours. Monitor the reaction completion by TLC. Filter the reaction mixture directly, and wash the filter cake with 10 mL of methanol and 10 mL of ethyl acetate successively to obtain 175 mg of light green solid compound F2 (E)-1,5-dimethyl-6-(2,4,6-trimethoxystyryl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, with a yield of 43.75%, melting point: 201 - 202 °C;

[0205] 1 H NMR (500 MHz, DMSO) δ 7.99 (s, 1H), 6.77 (d, J = 12.4 Hz, 1H), 6.50 (d, J = 12.4 Hz, 1H), 6.12 (s, 2H), 3.74 (s, 3H), 3.59 (s, 3H), 3.46 (s, 6H), 3.45 (s, 3H). HRMS (ESI) m / z: 357.1554.

[0206] Example 33

[0207] Preparation of compound (E)-6-(4-hydroxy-3,5-dimethoxystyryl)-1,5-dimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (F3):

[0208] Dissolve 50 mg (0.140 mmol) of compound F1 in 5 mL of acetic acid solution containing 40% hydrobromic acid, and reflux the reaction mixture at 90 °C for 1 hour. Monitor the reaction completion by TLC. Add 100 mL of water to the reaction mixture, filter after the formation of solid precipitate, and wash the filter cake with 3 mL of methanol twice to obtain 21 mg of yellow solid compound F3 (E)-6-(4-hydroxy-3,5-dimethoxystyryl)-1,5-dimethyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one, with a yield of 39.1%, melting point: 226 - 227 °C;

[0209] 1 H NMR (600 MHz, DMSO) δ 8.03 (s, 1H), 8.02 (s, 1H), 7.93 (d, J = 15.3 Hz, 1H), 7.30 (d, J = 15.2 Hz, 1H), 7.14 (s, 2H), 3.95 (s, 3H), 3.86 (s, 6H), 3.68 (s, 3H). HRMS (ESI) m / z: 343.1395.

[0210] Example 34

[0211] The tricyclic pyrazolopyrimidine aryl esters derivatives obtained in Example 4 - 33 were subjected to in vitro anti - tumor activity screening:

[0212] MTT method was used to detect cell viability:

[0213] For the cells in the logarithmic growth phase, the culture medium was aspirated, washed once with PBS, digested with trypsin, the digestion was terminated by adding culture medium, gently pipetted, counted, and inoculated in a 96 - well plate at the corresponding cell density (100 μL / well) for overnight culture. Then compounds were added (20 μL / well). A concentration gradient was set for each compound, and three replicates were set for each concentration. CO 2 The cells were cultured in an incubator at 37 °C for 48 hours. The old culture medium was aspirated and discarded, 100 μL of MTT was added, and then cultured for another 2 h. After incubating at 37 °C for 2 hours, the light absorption value (OD) at 570 nm was measured using an MB microplate reader;

[0214] Calculation formula:

[0215] Percentage of cell viability % = (OD of compound - OD of blank / OD of control - OD of blank) × 100%

[0216] Cell inhibition rate % = 1 - cell viability % = [1 - (OD of compound - OD of blank / OD of control - OD of blank)] × 100%. Using Graphpad, the IC 50 ;

[0217] Sample treatment: The samples were dissolved in dimethyl sulfoxide (DMSO) and stored at low temperature. The concentration of DMSO in the final system was controlled within the range that does not affect the detection of activity;

[0218] Data processing and result description: For the primary screening, under single - concentration conditions, for example, the monomer compound concentration was 50 μM; the extract was 50 μg / μL, and the activity of the samples was tested. For the samples that showed activity under certain conditions, such as the inhibition rate % (Inhibition) being greater than 70%, the dose - dependent relationship of the activity was further tested, that is, the IC 50 value, which was obtained by non - linear fitting of the sample concentration with the sample activity. The software used for calculation was Graphpad Prism 4; Usually, each sample was set with replicates (n ≥ 3) in the test.

[0219] Table 1 Results of in vitro anti - tumor biological activities of derivatives D1 - D23, E1 - E3, F1 - F3

[0220]

[0221]

[0222] As can be seen from the table: Using doxorubicin (DOX) as the positive control drug, all 29 compounds have good anti-cancer activity (IC 50 <5 μM), among which 24 compounds show excellent activity in inhibiting HCT116 colon cancer cells (IC 50 <1 μM), especially D2, D3, D4, D9, D11, D14, and E3 have the best activity (IC 50 = 0.01 μM). Among them, 26 compounds show excellent activity in inhibiting HGC27 gastric cancer cells (IC 50 <1 μM), especially D3, D7, D8, D9, D15, and E3 have the best activity (IC 50 equal to or less than 0.05 μM).

Claims

1. A tricyclic pyrazolopyrimidine aryl ester derivative, characterized in that the structure of the derivative is: wherein: Compound D1 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pentanoate; Compound D2 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl dodecanoate; Compound D3 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl isobutyrate; Compound D4 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl pivalate; Compound D5 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzoate; Compound D6 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethyl)benzoate; Compound D7 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzoate; Compound D8 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-fluorobenzoate; Compound D9 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl cyclopropanecarboxylate; Compound D10 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzoate; Compound D11 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-methylbenzoate; Compound D12 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-fluorobenzoate; Compound D13 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-chlorobenzoate; Compound D14 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-chlorobenzoate; Compound D15 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 3-bromobenzoate; Compound D16 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzoate; Compound D17 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 2-(trifluoromethyl)benzoate; Compound D18 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-nitrobenzenesulfonate; Compound D19 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-fluorobenzenesulfonate; Compound D20 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(trifluoromethoxy)benzenesulfonate; Compound D21 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-bromobenzenesulfonate; Compound D22 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-cyanobenzenesulfonate; Compound D23 is (E)-2,6-dimethoxy-4-((1-methyl-4-oxo-1,4,7,8-tetrahydropyrazolo[3,4-d]pyrimido[1,2-a]pyrimidin-9(6H)-ylidene)methyl)phenyl 4-(tert-butyl)benzenesulfonate; 2. Use of compound D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D20, D21, D22, D23 in the tricyclic pyrazolopyrimidine arylidene ester derivatives as claimed in claim 1 for the preparation of a medicament for treating HCT116 colon cancer.

3. Use of compound D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D20, D21, D22, D23 in the tricyclic pyrazolopyrimidine arylidene ester derivatives as claimed in claim 1 for the preparation of a medicament for treating HGC27 gastric cancer.

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