CN114478549B

CN114478549B - Pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivative and application thereof

Info

Publication number: CN114478549B
Application number: CN202210149626.3A
Authority: CN
Inventors: 阿吉艾克拜尔·艾萨; 祖克拉·肉孜; 聂礼飞; 胡尔西地·博扎罗夫; 赵江瑜
Original assignee: Xinjiang Technical Institute of Physics and Chemistry of CAS
Current assignee: Xinjiang Technical Institute of Physics and Chemistry of CAS
Priority date: 2022-02-18
Filing date: 2022-02-18
Publication date: 2023-06-23
Anticipated expiration: 2042-02-18
Also published as: CN114478549A

Abstract

The invention relates to pyrazolo [3,4 ]d]Pyrrolo [1, 2-)a]Pyrimidinone arylene derivative and application thereof, wherein the derivative takes 1-methyl (phenyl) group-5 amino-4 pyrazolo ethyl formate as an initial raw material, and reacts with pyrrolidone and valerolactam respectively under the action of phosphorus oxychloride to generate a ring to obtain 1-methyl (phenyl) group-dihydro pyrazolo [3,4 ]d]Pyrrolo [1,2 ]a]Pyrimidine-4 (6)H) Ketones (A and C) and 1-methyltetrahydropyrazole [3,4- ]d]Pyridine [1,2 ]a]Pyrimidine-4 (1)H) Ketone (E) and then reacted with differently substituted aromatic aldehydes and heterocyclic aromatic aldehydes under ethanolic sodium hydroxide conditions to form pyrazolo [3,4- ]d]Pyrrolo [1, 2-)a]Pyrimidinone arylene compounds. The cell activity screening result shows that: 29 compounds have inhibitory activity on HGC-27 human gastric cancer cells; 30 compounds have inhibitory activity against HT-29 human colon cancer cells; 16 compounds have inhibitory activity against HCT-116 human colon cancer cells; 21 compounds have inhibitory activity on HeLa cervical cancer cells; 19 compounds have inhibitory activity against ISK human endometrial cancer cells; there are 18 compounds that have inhibitory activity against MDA-MB-231 human breast cancer cells.

Description

Pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivative and application thereof

Technical Field

The invention relates to pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives and uses thereof.

Background

Cancer has become one of the most serious diseases threatening the health of the chinese population and causing death, and according to the latest statistics, malignant tumor death accounts for 23.91% of all the causes of death of residents. In 2019, 1 month, the national cancer center issues the latest national cancer statistics, and more than 1 ten thousand people are diagnosed as cancer every day on average, and compared with historical data, the cancer burden is in a continuous rising state. Medical costs for malignant tumors are over 2200 billions each year. Therefore, the prevention and treatment of tumors is becoming more and more severe, and how to effectively treat tumors is becoming urgent.

Anti-tumor drug studies began in the 40 s of the 20 th century and have been 70 years old so far. At present, traditional cytotoxicity medicines (such as paclitaxel and docetaxel) are still the main body of tumor drug treatment, and have the main defects of poor curative effect and large side effect on solid tumors. The focus of tumor drug development gradually changes from traditional cytotoxic drugs to novel targeting drugs based on molecular mechanisms, and the molecular targeting treatment of tumors achieves remarkable curative effects clinically. Pharmacy and oncologists are increasingly aware that they must start with the molecular mechanisms of tumor pathogenesis and design drugs for these specific molecules to improve efficacy and make breakthrough progress. Therefore, the research and development of the novel anti-tumor drug with high efficiency and low toxicity aiming at the molecular target is a key technical problem for urgent solving of tumor treatment, and also brings great social benefit and economic benefit.

Pyrimidine or pyrimidinone fragments are an indispensable class of important molecules in the field of life sciences because they can constitute various bases due to their unique structure. If pyrimidine is combined with five-membered heterocyclic ring, it will provide a wider range of nitrogen-containing groups during drug design and development.

The subject group is engaged in containing thiophene ring for a long time ^] And (3) synthesizing pyrimidine compounds and researching biological activity. M. Chauhan et al substituted thiophene ring with pyrazole group and synthesized a series of novel tricyclic pyrazolo [3,4-d ]]Pyrimidine derivatives, found pyrazolo [3,4-d ]]Pyrimidine compounds have various biological activities such as antibiosis, anti-inflammation, antivirus and the like, and meanwhile, the enzyme inhibition activity screening discovers the pyrazolo [3,4-d ]]Pyrimidine compounds are also useful as various kinase inhibitors. Most of the currently reported PDE9A inhibitors in phase II clinical trials of Alzheimer's disease have been completed and all contain a pyrazolopyrimidinone backbone.

Therefore, the pyrazolone [3,4-d ] pyrimidinone compound has great potential in the aspect of anti-tumor drug research and development and has broad market prospect. The biological activity of the oxazazole pyrimidine derivatives and the thiazole pyrimidine derivatives can be seen, and the synthetic method for searching and exploring the compounds has great theoretical and application values for searching new lead compounds such as medicines.

Reference is made to:

the cancer prevention and cure is a great emphasis in healthy Chinese construction [ J ]. Chinese medical guide, 2019,1 (8): 13-14.

Wood, liu Ming. Omnibearing daemon of anticancer pioneer [ J ]. Chinese sanitary talent 2019,4 (5): 12-13.

W.Chen; sun; zheng; h.zeng; S.Zhang; xia; yang; he Li; zou; J.He. Chinese cancer research (English edition) 2018,30 (1): 1-12.

Zheng Rongshou analysis of epidemic situation of Chinese malignant tumor in 2015 [ J ]. Chinese tumor, 2019,41 (1): 19-28.

Zhao Hui design, synthesis and biological evaluation of novel CDK/6 inhibitors [ D ] university of Zhejiang paper 2019.

Zhang Yan, yinxi, medical improvement, "old questions" and New prescription [ J ]. Chinese economic cycle, 2019,5:23-27.

Fang Luo, wu Yingying, zhang. Current global anti-tumor drug development in recent 5 years [ J ]. Chinese tumor, 2013,07:51-58.

Hanahan D.Rethinking the war on cancer[J].The Lancet.2014,383(9916):558-563.

a) K.Bozorov, J.Y.Zhao, B.Elmuradov, A.Pataer, H.A.Aisa, european Journal of Medicinal Chemistry,2015,102:552-573; b) K.Bozorov, J.Y.Zhao, H.A.Aisa, arkivoc,2017,41-66; c) K.Bozorov, L.F.Nie, J.Y.Zhao, H.A.Aisa, european Journal of Medicinal Chemistry,2017,140:465-493; d) L.F.Nie, K.Bozorov, C.Niu, G.Z.Huang, H.A.Aisa, research on Chemical Intermediates,2017,43:6835-6843; e) L.F.Nie, G.Z.Huang, K.Bozorov, J.Y.Zhao, C.Niu, S.S.Sagdullaev, H.A.Aisa, heterocyclic Communications,2018,24:43-50; f) L.F.Nie, K.Bozorov, G.Huang, J.Zhao, C.Niu, H.A.Aisa, phosphorus, sulfur, and Silicon and the Related Elements2018,193,656-667; g) H.A.Aisa, K.Bozorov, J.Zhao, C.Niu, K.Shakhidoyatov, chinese patent 2016,CN 104016963B

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V.A.Makarov,H.Braun,M.Richter,O.B.Riabova,J.Kirchmair,E.S.Kazakova,N.Seidel,P.Wutzler,M.Schmidtke,Chem.Med.Chem.,2015,10:1629-1634.

a)G.-B.Li,S.Ma,L.-L.Yang,S.Ji,Z.Fang,G.Zhang,L.-J.Wang,J.-M.Zhong,Y.Xiong,J.-H.Wang,S.-Z.Huang,L.-L.Li,R.Xiang,D.Niu,Y.-C.Chen,S.-Y.Yang,Journal of Medicinal Chemistry,2016,59,8293-8305；b)S.Schenone,M.Radi,F.Musumeci,C.Brullo,M.Botta,Chemical Reviews,2014,114:7189-7238.

Based on comprehensive analysis of related patents and documents at home and abroad, pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene compounds are fully synthesized, simple transformation and modification are carried out, phenyl and heterocycle containing different substituents are introduced into pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene compound molecules, so that the patentability of the pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene compounds is improved, and candidate drugs with remarkable curative effects and definite target antitumor activity on HGC-27 human gastric cancer cells, HT-29 human colon cancer cells, HCT-116 human colon cancer cells, heLa cervical cancer cells, ISK human endometrial cancer cells and MDA-MB-231 human breast cancer cells are researched. The activity screening results show that: 29 compounds have inhibitory activity on HGC-27 human gastric cancer cells; 30 compounds have inhibitory activity against HT-29 human colon cancer cells; 16 compounds have inhibitory activity against HCT-116 human colon cancer cells; 21 compounds have inhibitory activity on HeLa cervical cancer cells; 19 compounds have inhibitory activity against ISK human endometrial cancer cells; there are 18 compounds that have inhibitory activity against MDA-MB-231 human breast cancer cells.

Disclosure of Invention

The invention aims to provide pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives and application thereof. The compound takes 1-methyl (benzene) group-5 amino-4 pyrazolo ethyl formate as an initial raw material, and respectively reacts with pyrrolidone and valerolactam under the action of phosphorus oxychloride to form a ring to obtain 1-methyl (benzene) group-dihydro pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -ketone (A and C) and 1-methyl tetrahydropyrazolo [3,4-D ] pyridine [1,2-a ] pyrimidine-4 (1H) -ketone (E), and then reacts with different substituted aromatic aldehyde and heterocyclic aromatic aldehyde under the condition of ethanol sodium hydroxide to generate pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene compounds (B1-B14, D1-D14 and F1-F14). The cell activity screening result shows that: 29 compounds have inhibitory activity on HGC-27 human gastric cancer cells; 30 compounds have inhibitory activity against HT-29 human colon cancer cells; 16 compounds have inhibitory activity against HCT-116 human colon cancer cells; 21 compounds have inhibitory activity on HeLa cervical cancer cells; 19 compounds have inhibitory activity against ISK human endometrial cancer cells; there are 18 compounds that have inhibitory activity against MDA-MB-231 human breast cancer cells. The structural formula of the pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivative is as follows:

Wherein:

compound a is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

the compound B1 is (E) -1-methyl-8-benzylidene-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

the compound B2 is (E) -1-methyl-8- (3-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

the compound B3 is (E) -1-methyl-8- (4-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

the compound B4 is (E) -1-methyl-8- (3-chlorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B5 is (E) -1-methyl-8- (4-fluorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B6 is (E) -1-methyl-8- (4-methoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B7 is (E) -1-methyl-8- (3, 4, 5-trimethoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B8 is (E) -1-methyl-8- (thiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B9 is (E) -1-methyl-8- (4, 5-dimethylthiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

Compound B10 is (E) -1-methyl-8- (furan-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B11 is (E) -1-methyl-8- (5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B12 is (E) -1-methyl-8- (4, 5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B13 is (E) -1-methyl-8- (2, 3-dihydrobenzo [ B ] [1,4] dioxin-6-ylmethylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound B14 is (E) -1-methyl-8- (ferrocene-2-methylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound C is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D1 is (E) -1-phenyl-8-benzylidene-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D2 is (E) -1-phenyl-8- (3-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D3 is (E) -1-phenyl-8- (4-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

Compound D4 is (E) -1-phenyl-8- (3-chlorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D5 is (E) -1-phenyl-8- (4-fluorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D6 is (E) -1-phenyl-8- (4-methoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D7 is (E) -1-phenyl-8- (3, 4, 5-trimethoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D8 is (E) -1-phenyl-8- (thiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D9 is (E) -1-phenyl-8- (4, 5-dimethylthiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D10 is (E) -1-phenyl-8- (furan-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D11 is (E) -1-phenyl-8- (5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D12 is (E) -1-phenyl-8- (4, 5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

Compound D13 is (E) -1-phenyl-8- (2, 3-dihydrobenzo [ b ] [1,4] dioxin-6-ylmethylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound D14 is (E) -1-phenyl-8- (ferrocene-2-methylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one;

compound E is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

the compound F1 is (E) -1-methyl-9-benzylidene-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidin-4 (1H) -one;

the compound F2 is (E) -1-methyl-9- (2-methylbenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidin-4 (1H) -one;

compound F3 is (E) -1-methyl-9- (4-methylbenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F4 is (E) -1-methyl-9- (3-chlorobenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F5 is (E) -1-methyl-9- (4-fluorobenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F6 is (E) -1-methyl-9- (4-methoxybenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F7 is (E) -1-methyl-9- (3, 4, 5-trimethoxybenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

Compound F8 is (E) -1-methyl-9- (thiophen-2-yl) methylene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F9 is (E) -1-methyl-9- (4, 5-dimethylthiophen-2-yl) methylene) -6,7,8, 9-tetrahydropyrazol [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F10 is (E) -1-methyl-9- (furan-2-yl) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F11 is (E) -1-methyl-9- (5-dimethylfuran-2-yl) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

compound F12 is (E) -1-methyl-9- (4, 5-dimethylfuran-2-yl) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidin-4 (1H) -one;

compound F13 is (E) -1-methyl-9- (2, 3-dihydrobenzo [ b ] [1,4] dioxin-6-ylmethylene) -6,7,8, 9-tetrahydropyrazol [3,4-d ] pyridin [1,2-a ] pyrimidin-4 (1H) -one;

the compound F14 is (E) -1-methyl-9- (ferrocene-2-methylene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidin-4 (1H) -one.

The application of the compounds B2, B7, B8, B10, B12, D2-D13, D14 and F1-F13 in preparing medicines for treating HGC-27 human gastric cancer in pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives.

The application of the compounds B2, B7, B8, B12, B13, D2-D12, D14 and F1-F14 in preparing medicines for treating HT-29 human colon cancer in pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives.

The application of the compounds B2, B7, D2-D4, F1-F9, F12 and F13 in pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives in preparing medicaments for treating HCT-116 human colon cancer.

The application of the compounds B1, B2, B4, B7, B10, D2, D3, D5, D7, D10, F1-F4, F6-F9 and F11-F13 in preparing the medicines for treating Hela cervical cancer is provided.

The pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives have the application of compounds B1, B2, B4, B7, B10, D2-D5, D7, D10, F1, F2, F4, F6, F7, F9, F11-F13 in preparing medicaments for treating ISK human endometrial cancer.

The application of the compounds B2, B7, B12, D2-D5, D10, F1-F5, F7, F9 and F11-F13 in preparing medicines for treating MDA-MB-231 human breast cancer in pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives.

The invention relates to pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives and application thereof, wherein the preparation method of the pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives is carried out according to the following steps:

Preparation of Compound A:

dissolving 1.69g of compound 1-methyl-5 amino-4 ethyl pyrazole formate, 10mmoL in 20mL of anhydrous dioxane, adding 1.3g of pyrrolidone and 15mmoL, slowly dropwise adding 3.8g of phosphorus oxychloride and 25mmoL, carrying out reflux reaction until all raw materials disappear, filtering a reaction solution, concentrating, and carrying out gradient elution by adopting a forward silica gel column chromatography, wherein an eluent is petroleum ether and ethyl acetate in a volume ratio of 1:2 to obtain a compound A;

preparation of Compounds B1-B14:

dissolving a compound A1.9g and 10mmoL in a 20mL mixed solution of absolute ethyl alcohol and sodium hydroxide 1.2g and 30mmoL, slowly dripping aldehyde 15mmoL with different substitutions, carrying out reflux reaction until all raw materials disappear, filtering a reaction solution, concentrating, and carrying out gradient elution by adopting a forward silica gel column chromatography, wherein an eluent is pure ethyl acetate to obtain a compound B1-B14;

preparation of compound C:

dissolving 2.3g of compound 1-phenyl-5 amino-4 ethyl pyrazole formate, 10mmoL in 20mL of anhydrous dioxane, adding 1.3g of pyrrolidone and 15mmoL, slowly dropwise adding 3.8g of phosphorus oxychloride and 25mmoL, carrying out reflux reaction until all raw materials disappear, filtering reaction liquid, concentrating, adopting forward silica gel column chromatography for gradient elution, wherein an eluent is petroleum ether and ethyl acetate in a volume ratio of 1:2, and obtaining a compound C;

Preparation of Compounds D1-D14:

dissolving 2.5g of compound C, 10mmoL in a 20mL mixed solution of anhydrous ethanol and 1.2g of sodium hydroxide, and 30mmoL, slowly dripping 15mmoL of aldehyde with different substitutions, refluxing and reacting until all raw materials disappear, filtering the reaction solution, concentrating, and performing gradient elution by adopting a forward silica gel column chromatography, wherein an eluent is pure ethyl acetate to obtain compounds D1-D14;

preparation of Compound E:

dissolving 1.69g of compound 1-methyl-5 amino-4 pyrazolecarboxylic acid ethyl ester, 10mmoL in 20mL of anhydrous dioxane, adding 1.5g of valerolactam and 15mmoL, slowly dropwise adding 3.8g of phosphorus oxychloride and 25mmoL, carrying out reflux reaction until all raw materials disappear, filtering the reaction solution, concentrating, and carrying out gradient elution by adopting a forward silica gel column chromatography, wherein an eluent is petroleum ether and ethyl acetate in a volume ratio of 1:1 to obtain a compound E;

preparation of Compounds F1-F14:

dissolving compound E2 g,10mmoL in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, slowly dripping different substituted aldehydes 15mmoL, refluxing until the raw materials completely disappear, filtering the reaction solution, concentrating, and performing gradient elution by adopting a forward silica gel column chromatography, wherein an eluent is pure ethyl acetate, thus obtaining the compounds F1-F14.

The pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -ketone (A and C) and 1-methyltetrahydrophyrazol [3,4-D ] pyridine [1,2-a ] pyrimidine-4 (1H) -ketone (E) are prepared by respectively reacting ethyl 1-methyl (benzene) yl-5 amino-4 pyrazoloformate serving as an initial raw material with pyrrolidone and valerolactam under the action of phosphorus oxychloride to form a ring to obtain pyrazolo [3,4-D ] pyrrole [1,2-a ] pyrimidine-4 (6H) -ketone (A and C) and 1-methyltetrahydrophyrazol [3,4-D ] pyridine [1,2-a ] pyrimidine-4 (1H) -ketone (E), and then reacting with different substituted aromatic aldehydes and heterocyclic aromatic aldehydes under the condition of ethanol sodium hydroxide to obtain pyrazolo [3,4-D ] pyrrole (pyridine) and [1,2-a ] pyrimidine-ketone arylene compound (B1-B14, D1-D14, F1-F14), wherein the synthetic route is as follows:

the pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives provided by the invention have the inhibitory activity of 42 obtained compounds on HGC-27 human gastric cancer cells, HT-29 human colon cancer cells, HCT-116 human colon cancer cells, heLa cervical cancer cells, ISK human endometrial cancer cells and MDA-MB-231 human breast cancer cells, and the results show that: the compounds B2, B7, B8, B10, B12, D2-D13, D14, F1-F13 have inhibitory activity on HGC-27 human gastric cancer cells; compounds B2, B7, B8, B12, B13, D2-D12, D14, F1-F14 have inhibitory activity against HT-29 human colon cancer cells; compounds B2, B7, D2-D4, F1-F9, F12, F13 have inhibitory activity against HCT-116 human colon cancer cells; the compounds B1, B2, B4, B7, B10, D2, D3, D5, D7, D10, F1-F4, F6-F9, F11-F13 have inhibitory activity on HeLa cervical cancer cells; compounds B1, B2, B4, B7, B10, D2-D5, D7, D10, F1, F2, F4, F6, F7, F9, F11-F13 have inhibitory activity against ISK human endometrial cancer cells; compounds B2, B7, B12, D2-D5, D10, F1-F5, F7, F9, F11-F13 have inhibitory activity against MDA-MB-231 human breast cancer cells.

Detailed Description

The present invention is further illustrated by examples, but the present invention is not limited to these examples;

reagent: all reagents were commercially available in analytical purity;

example 1

Preparation of Compound A:

dissolving 1.69g of compound 1-methyl-5 amino-4 pyrazolo ethyl formate and 10mmoL in 20mL of anhydrous dioxane, adding 1.3g of pyrrolidone and 15mmoL, slowly dropwise adding 3.8g of phosphorus oxychloride and 25mmoL, refluxing until all raw materials disappear, filtering the reaction liquid, concentrating, and gradient eluting by adopting a forward silica gel column chromatography, wherein an eluent is petroleum ether and ethyl acetate in a volume ratio of 1:2 to obtain a compound A which is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -ketone, and the yield is as follows: 75%, white solid, mp 202-203 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.02(1H,s,H3),4.16(2H,dd,J＝7.2,7.4Hz,H6),3.96(3H,s,NCH ₃ ),3.16(2H,t,J＝8.0Hz,H8),2.31(2H,m,H7)。

example 2

Preparation of compound B1:

the compound A obtained in example 1 was 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyr-rolidin-4 (6H) -one 2.5g,10mmoL was dissolved in a 20mL mixed solution of absolute ethanol and sodium hydroxide 1.2g,30mmoL, then benzaldehyde 1.6g,15mmoL was slowly added dropwise thereto, the reaction solution was refluxed until the starting material was completely disappeared, the reaction solution was filtered, concentrated, and eluted with a forward silica gel column chromatography gradient using pure ethyl acetate as an eluent to give the compound B1 as (E) -1-methyl-8-benzylidene-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, yield: 80%, white solid, melting point: 242-244 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.02(1H,s,H3),7.79(1H,t,J＝2.9Hz,H7′),7.54(2H,d,J＝7.5Hz,H2′and H6′),7.45(2H,t,J＝7.5Hz,H3′and H5′),7.38(1H,t,J＝7.3Hz,H4′),4.21(2H,dd,J＝7.7,6.5Hz,H6),4.02(3H,s,NCH ₃ ),3.29(2H,ddd,J＝7.4,6.6,2.8Hz,H7)。

Example 3

Preparation of compound B2:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 3-methylbenzaldehyde 1.8g,15mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate, so that the compound B2 is (E) -1-methyl-8- (3-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 81%, pale yellow solid, melting point: 250-252 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.05(1H,s,H3),7.79(1H,t,J＝2.8Hz,H7′),7.36(3H,m,H2′,H4′and H5′),7.21(1H,dt,J＝6.5,1.8Hz,H6′),4.24(2H,dd,J＝7.89,6.6Hz,H6),4.03(3H,s,NCH ₃ ),3.32(2H,ddd,J＝7.9,6.6,2.8Hz,H7),2.41(3H,s,CH ₃ 3′)。

example 4

Preparation of compound B3:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 4-methylbenzaldehyde 1.8g,15mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate, so that the compound B3 is (E) -1-methyl-8- (4-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 79%, pale yellow solid, melting point: 210-212 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.03(1H,s,H3),7.78(1H,t,J＝2.8Hz,H7′),7.45(2H,d,J＝8.3Hz,H2′and H6′),7.26(2H,d,J＝8.3Hz,H3′and H5′),4.22(2H,dd,J＝7.7,6.7Hz,H6),4.02(3H,s,NCH ₃ ),3.29(2H,ddd,J＝7.4,6.7,2.8Hz,H7)。

Example 5

Preparation of compound B4:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 3-chlorobenzaldehyde 2.1g,15mmoL is slowly added dropwise, the reflux reaction is carried out until the raw materials completely disappear, the reaction solution is filtered, concentrated, and eluted by a forward silica gel column chromatography gradient with pure ethyl acetate as eluent to obtain the compound B4 which is (E) -1-methyl-8- (3-chlorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, the yield: 70%, pale yellow solid, melting point: 266-268 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.07(1H,s,H3),7.76(1H,t,J＝2.9Hz,H7′),7.55(1H,br.s,H2′),7.44(1H,dt,J＝7.3,1.7Hz,H4′),7.40(1H,dd,J＝7.6,7.3Hz,H5′),7.37(1H,dt,J＝7.6,1.8Hz,H6′),4.27(2H,dd,J＝7.4,6.9Hz,H6),4.04(3H,s,NCH ₃ ),3.34(2H,ddd,J＝7.4,6.9,2.9Hz,H7)。

example 6

Preparation of compound B5:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 4-fluorobenzaldehyde 1.86g,15mmoL is slowly added dropwise, the reaction solution is filtered and concentrated until the raw materials completely disappear, and the eluent is pure ethyl acetate by adopting a forward silica gel column chromatography gradient elution to obtain the compound B5 which is (E) -1-methyl-8- (4-fluorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 69%, white solid, melting point: 268-270 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.05(1H,s,H3),7.79(1H,t,J＝2.8Hz,H7′),7.55(2H,dd,J＝8.7,5.4Hz,H2′and H6′),7.16(2H,t,J＝8.6Hz,H3′and H5′),4.26(2H,dd,J＝7.8,6.7Hz,H6),4.03(3H,s,NCH ₃ ),3.30(2H,ddd,J＝7.8,6.7,2.8Hz,H7)。

Example 7

Preparation of compound B6:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 4-methoxybenzaldehyde 2.0g,15mmoL is slowly added dropwise, the reflux reaction is carried out until the raw materials completely disappear, the reaction solution is filtered, concentrated, and eluted by a forward silica gel column chromatography gradient with pure ethyl acetate as eluent to obtain the compound B6 which is (E) -1-methyl-8- (4-methoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, the yield: 85%, pale yellow solid, melting point: 215-217 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.77(1H,t,J＝2.8Hz,H7′),7.52(2H,d,J＝8.8Hz,H2′and H6′),6.98(2H,d,J＝8.8Hz,H3′and H5′),4.23(2H,dd,J＝7.9,6.6Hz,H6),4.03(3H,s,NCH ₃ ),3.87(3H,s,OCH ₃ 4′),3.29(2H,ddd,J＝7.9,6.6,2.8Hz,H7)。

example 8

Preparation of compound B7:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 3,4, 5-trimethoxybenzaldehyde 2.9g,15mmoL is slowly added dropwise, the reaction solution is refluxed until the raw materials completely disappear, the reaction solution is filtered and concentrated, and the pure ethyl acetate is used for elution by a forward silica gel column chromatography gradient, so that the compound B7 is (E) -1-methyl-8- (3, 4, 5-trimethoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is obtained: 87%, pale yellow solid, melting point: 273-275 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.05(1H,s,H3),7.74(1H,t,J＝2.8Hz,H7′),6.79(2H,s,H2′and H6′),4.26(2H,dd,J＝7.8,6.6Hz,H6),4.04(3H,s,NCH ₃ ),3.93(OCH ₃ 3′and 5′),3.92(3H,s,OCH ₃ 4′),3.33(2H,ddd,J＝7.4,6.6,2.8Hz,H7)。

Example 9

Preparation of compound B8:

2.5g of compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 1.68g of thiophene-2-formaldehyde and 15mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and gradient elution is carried out by adopting a forward silica gel column chromatography and pure ethyl acetate is adopted as an eluent, so that the compound B8 is (E) -1-methyl-8- (thiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -one, and the yield is as follows: 76%, pale yellow solid, melting point: 238-240 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),8.02(1H,t,J＝2.8Hz,H6′),7.56(1H,d,J＝5.1Hz,H5′),7.37(1H,d,J＝3.7Hz,H3′),7.19(1H,dd,J＝5.1,3.7Hz,H4′),4.28(2H,dd,J＝7.9,6.6Hz,H6),4.02(3H,s,NCH ₃ ),3.22(2H,ddd,J＝7.3,6.6,2.8Hz,H7)。

example 10

Preparation of compound B9:

the compound A obtained in example 1 was 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrimidin-4 (6H) -one 2.5g,10mmoL was dissolved in a mixed solution of 20mL of anhydrous ethanol and sodium hydroxide 1.2g,30mmoL, then 5-methylthiophene-2-carbaldehyde 1.89g,15mmoL was slowly added dropwise, the reaction was refluxed until the starting materials disappeared completely, the reaction solution was filtered and concentrated, and the elution was performed by using a forward silica gel column chromatography gradient, and the eluent was pure ethyl acetate, to obtain compound B9 as (E) -1-methyl-8- (4, 5-dimethylthiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, yield: 73%, orange solid, melting point: 292-294 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.03(1H,s,H3),7.86(1H,t,J＝2.8Hz,H6′),7.06(1H,s,H3′),4.25(2H,dd,J＝8.0,6.6Hz,H6),4.01(3H,s,NCH ₃ ),3.16(2H,m,H7),2.42(3H,s,CH ₃ 5′),2.18(3H,s,CH ₃ 4′)。

Example 11

Preparation of compound B10:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then furan-2-formaldehyde 1.44g,15mmoL is slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate, so that the compound B10 is (E) -1-methyl-8- (furan-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 82%, pale yellow solid, melting point: 248-250 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.59(1H,d,J＝1.8Hz,H5′),7.58(1H,t,J＝2.8Hz,H6′),6.66(1H,d,J＝3.4Hz,H3′),6.55(1H,dd,J＝3.4,1.8Hz,H4′),4.24(2H,dd,J＝7.6,7.0Hz,H6),4.01(3H,s,NCH ₃ ),3.35(2H,td,J＝7.5,2.8Hz,H7)。

example 12

Preparation of compound B11:

the compound a obtained in example 1 was 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmol was dissolved in a 20mL mixed solution of absolute ethanol and sodium hydroxide 1.2g,30mmol, then 5-methylfuran-2-carbaldehyde 1.65g,15mmol was slowly added dropwise, the reaction was refluxed until the starting materials disappeared completely, the reaction solution was filtered and concentrated, and the eluent was pure ethyl acetate by a forward silica gel column chromatography gradient to give compound B11 as (E) -1-methyl-8- (5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, yield: 79%, pale yellow solid, melting point: 219-221 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.03(1H,s,H3),7.51(1H,t,J＝2.7Hz,H6′),6.57(1H,d,J＝3.3Hz,H3′),6.16(1H,dd,J＝3.3,1.1Hz,H4′),4.22(2H,dd,J＝7.9,6.7Hz,H6),3.99(3H,s,NCH ₃ ),3.31(2H,td,J＝7.4,2.7Hz,H7),2.40(3H,s,CH ₃ 5′)。

Example 13

Preparation of compound B12:

the compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL is dissolved in a 20mL anhydrous ethanol and sodium hydroxide 1.2g,30mmoL mixed solution, then 4, 5-dimethylfuran-2-formaldehyde 1.86g,15mmoL is slowly added dropwise, reflux reaction is carried out until all the raw materials disappear, the reaction solution is filtered and concentrated, forward silica gel column chromatography gradient elution is adopted, eluent is pure ethyl acetate, and the compound B12 is (E) -1-methyl-8- (4, 5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, the yield: 83%, pale yellow solid, melting point: 257-259 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.47(1H,t,J＝2.7Hz,H6′),6.47(1H,s,H3′),4.23(2H,dd,J＝7.2,7.5Hz,H6),4.00(3H,s,NCH ₃ ),3.30(2H,t,J＝7.4Hz,H7),2.30(3H,s,CH ₃ 5′),2.00(3H,s,CH ₃ 4′)。

example 14

Preparation of compound B13:

the compound A obtained in example 1 was 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmoL was dissolved in 20mL of a mixed solution of absolute ethanol and sodium hydroxide 1.2g,30mmoL, then 2, 3-dihydrobenzo [ B ] [1,4] dioxin-6-carbaldehyde 2.46g,15mmoL was slowly added dropwise thereto, the reaction was refluxed until all the starting materials disappeared, the reaction solution was filtered, concentrated, and the mixture was eluted using a forward silica gel column chromatography gradient, and the eluent was pure ethyl acetate to give compound B13 as (E) -1-methyl-8- (2, 3-dihydrobenzo [ B ] [1,4] dioxin-6-ylmethylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, yield: 74%, pale yellow solid, melting point: 262-264 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.69(1H,t,J＝2.8Hz,H7′),7.08(1H,dd,J＝2.1Hz,H2′),7.06(1H,dd,J＝8.3,2.1Hz,H6′),6.93(1H,d,J＝8.3Hz,H5′),4.31(4H,m,2xOCH ₂ dioxane),4.22(2H,dd,J＝7.9,6.7Hz,H6),4.02(3H,s,NCH ₃ ),3.27(2H,ddd,J＝7.9,6.7,2.8Hz,H7)。

Example 15

Preparation of compound B14:

2.5g of compound A obtained in example 1 is 1-methyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 3.36g of ferrocene formaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate, so that the compound B14 is (E) -1-methyl-8- (ferrocene-2-methylene) -7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -one, and the yield is obtained: 87%, pale yellow solid, melting point: 272-274 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.02(1H,s,H3),7.59(1H,br.s,H6′),4.55(2H,s,H2′and H5′),4.46(2H,s,H3′and H4′),4.19(5H,br.s,H1″-H5″),4.19(2H,overlapped,H6),4.01(3H,s,NCH ₃ ),3.01(2H,br.t,J＝8.3Hz,H7)。

example 16

Preparation of compound C:

2.3g of compound 1-phenyl-5 amino-4 pyrazolo ethyl formate and 10mmoL are dissolved in 20mL of anhydrous dioxane, 1.3g of pyrrolidone and 15mmoL are added, 3.8g of phosphorus oxychloride and 25mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction liquid is filtered, concentrated, gradient elution is carried out by adopting forward silica gel column chromatography, the eluent is petroleum ether and ethyl acetate in a volume ratio of 1:2, and the compound C is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -ketone, and the yield is: 73%, white solid, mp 206-207 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.21(1H,s,H3),8.04(2H,dd,J＝8.5,1.3Hz,H2′and H6′),7.50(2H,dd,J＝8.5,7.5Hz,H3′and H5′),7.34(1H,tt,J＝7.5,1.3Hz,H4′),4.16(2H,t,J＝7.3Hz,H6),3.18(2H,t,J＝8.0Hz,H8),2.31(2H,m,H7)。

Example 17

Preparation of compound D1:

2.5g of compound C obtained in example 16, namely 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyr-rolidin-4 (6H) -one, 10mmoL, is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, 1.6g of benzaldehyde and 15mmoL are slowly added dropwise, the mixture is refluxed until all the raw materials disappear, the reaction solution is filtered, concentrated and eluted by a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate, thus obtaining compound D1, namely (E) -1-phenyl-8-benzylidene-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 63%, melting point of pale yellow solid: 211-213 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):8.24(1H,s,H3),8.16(2H,dd,J＝8.7,1.2Hz,H2′and H6′),7.73(1H,t,J＝2.8Hz,H7″),7.54(2H,dd,J＝8.6,7.4Hz,H3′and H5′),7.35(4H,m,H4′,H2″,H4″,H5″),7.14(1H,m,H6″),4.23(2H,dd,J＝7.9,6.6Hz,H6),3.30(2H,ddd,J＝7.9,6.6,2.8Hz,H7)。

example 18

Preparation of compound D2:

2.5g of compound C obtained in example 16 is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one and 10mmoL are dissolved in 20mL of a mixed solution of absolute ethanol and sodium hydroxide of 1.2g and 30mmoL, then 1.8g of 3-methylbenzaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound D2 which is (E) -1-phenyl-8- (3-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 82%, melting point of pale yellow solid: 234-236 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.17(2H,dd,J＝8.7,1.2Hz,H2′and H6′),7.77(1H,t,J＝2.8Hz,H7″),7.54(2H,dd,J＝8.6,7.4Hz,H3′and H5′),7.36(4H,m,H4′,H2″,H4″,H5″),7.20(1H,m,H6″),4.26(2H,dd,J＝7.9,6.6Hz,H6),3.33(2H,ddd,J＝7.9,6.6,2.8Hz,H7),2.41(3H,s,CH ₃ 3″)。

Example 19

Preparation of compound D3:

2.5g of compound C obtained in example 16 is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 1.8g of 4-methylbenzaldehyde and 15mmoL are slowly added dropwise, the reaction solution is filtered and concentrated until all raw materials disappear, and the reaction solution is eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound D3 which is (E) -1-phenyl-8- (4-methylbenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 85%, pale yellow solid, melting point: 227-229 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.17(2H,dd,J＝8.6,1.3Hz,H2′and H6′),7.77(1H,t,J＝2.8Hz,H7″),7.54(2H,dd,J＝8.6,7.4Hz,H3′and H5′),7.45(2H,d,J＝8.2,Hz,H2″and H6″),7.36(1H,tt,J＝7.4,1.2Hz,H4′),7.26(2H,dd,J＝8.2Hz,H3″and H5″),4.26(2H,dd,J＝7.9,6.6Hz,H6),3.31(2H,ddd,J＝7.9,6.6,2.8Hz,H7),2.40(3H,s,CH ₃ 4″)。

example 20

Preparation of compound D4:

2.5g of compound C obtained in example 16, namely 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 2.1g of 3-chlorobenzaldehyde and 15mmoL are slowly added dropwise, the reaction solution is refluxed until all the raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound D4, namely (E) -1-phenyl-8- (3-chlorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 73%, pale yellow solid, melting point: 256-258 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.25(1H,s,H3),8.15(2H,dd,J＝8.7,1.2Hz,H2′and H6′),7.74(1H,t,J＝2.9Hz,H7″),7.55(2H,dd,J＝8.6,7.4Hz,H3′and H5′),7.53(1H,d,J＝1.8Hz,H2″),7.44(1H,dt,J＝7.3,1.8Hz,H4″),7.40(1H,dd,J＝7.7,7.3Hz,H5″),7.37(2H,m,H4′and H6″),4.30(2H,dd,J＝7.7,6.5Hz,H6),3.35(2H,ddd,J＝7.6,6.6,2.9Hz,H7)。

Example 21

Preparation of compound D5:

2.5g of compound C obtained in example 16 is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one and 10mmoL are dissolved in 20mL of a mixed solution of absolute ethanol and sodium hydroxide of 1.2g and 30mmoL, then 1.86g of 4-fluorobenzaldehyde and 15mmoL are slowly added dropwise, the reaction solution is filtered and concentrated until all raw materials disappear, and the reaction solution is eluted by a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound D5 which is (E) -1-phenyl-8- (4-fluorobenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 76%, pale yellow solid, melting point: 231-233 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.15(2H,dd,J＝8.7,1.2Hz,H2′and H6′),7.76(1H,t,J＝2.8Hz,H7″),7.54(4H,m,H3′,H5′,H2″and H6″),7.37(1H,tt,J＝7.4,1.2Hz,H4′),7.15(2H,t,J＝8.6Hz,H3″and H5″),4.28(2H,dd,J＝7.8,6.5Hz,H6),3.30(2H,ddd,J＝7.6,7.0,2.8Hz,H7)。

example 22

Preparation of compound D6:

2.5g of compound C obtained in example 16, namely 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, 10mmoL, is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 2.0g of 4-methoxybenzaldehyde and 15mmoL are slowly added dropwise, the reaction solution is refluxed until all the raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain a compound D6, namely (E) -1-phenyl-8- (4-methoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 84%, white solid, melting point: 249-251 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.17(2H,dd,J＝8.8,1.3Hz,H2′and H6′),7.75(1H,t,J＝2.7Hz,H7″),7.55(2H,d,J＝8.8,Hz,H2″and H6″),7.52(2H,dd,J＝8.8,7.3Hz,H3′and H5′),7.36(1H,tt,J＝7.4,1.2Hz,H4′),6.97(2H,dd,J＝8.8Hz,H3″and H5″),4.26(2H,dd,J＝7.7,6.9Hz,H6),3.87(OCH ₃ 4″),3.30(2H,ddd,J＝7.7,6.9,2.7Hz,H7)。

Example 23

Preparation of compound D7:

2.5g of compound C obtained in example 16 is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrimidine-4 (6H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 2.9g of 3,4, 5-trimethoxybenzaldehyde and 15mmoL are slowly added dropwise, the mixture is refluxed until all raw materials disappear, the reaction solution is filtered and concentrated, and the mixture is eluted by a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain compound D7 which is (E) -1-phenyl-8- (3, 4, 5-trimethoxybenzylidene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidine-4 (6H) -one, and the yield is as follows: 79%, pale yellow solid, melting point: 266-268 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.24(1H,s,H3),8.16(2H,dd,J＝8.7,1.1Hz,H2′and H6′),7.72(1H,t,J＝2.8Hz,H7″),7.55(2H,dd,J＝8.6,7.6Hz,H3′and H5′),7.38(1H,tt,J＝7.6,1.1Hz,H4′),6.78(2H,s,H2″and H6″),4.29(2H,dd,J＝8.2,6.6Hz,H6),3.93(OCH ₃ 3″and 5″),3.92(OCH ₃ 4″),3.35(2H,ddd,J＝8.0,7.0,2.7Hz,H7)。

example 24

Preparation of compound D8:

2.5g of compound C obtained in example 16 is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 1.68g of thiophene-2-formaldehyde and 15mmoL are slowly added dropwise, the reaction solution is filtered and concentrated until all raw materials disappear, and the reaction solution is eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound D8 which is (E) -1-phenyl-8- (thiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 69%, pale yellow solid, melting point: 282-284 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.24(1H,s,H3),8.16(2H,dd,J＝8.7,1.2Hz,H2′and H6′),8.01(1H,t,J＝2.8Hz,H6″),7.56(2H,m,H3′and H5′),7.55(1H,m,H5″),7.38(2H,m,H3″and H4′),7.19(1H,dd,J＝5.1,3.7Hz,H4″),4.32(2H,dd,J＝7.9,6.6Hz,H6),3.26(2H,ddd,J＝7.9,6.6,2.8Hz,H7)。

Example 25

Preparation of compound D9:

compound C obtained in example 16 was 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrimidin-4 (6H) -one 2.5g,10mmol was dissolved in a mixed solution of 20mL of absolute ethanol and sodium hydroxide 1.2g,30mmol, then 5-methylthiophene-2-carbaldehyde 1.89g,15mmol was slowly added dropwise, the reaction was refluxed until all the starting materials disappeared, the reaction solution was filtered, concentrated, and eluted with a forward silica gel column chromatography gradient using pure ethyl acetate as eluent to give compound D9 as (E) -1-phenyl-8- (4, 5-dimethylthiophen-2-yl) methylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, yield: 61%, pale yellow solid, melting point: 297-299 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.20(1H,s,H3),8.12(2H,dd,J＝8.7,1.2Hz,H2′and H6′),8.00(1H,t,J＝2.8Hz,H6″),7.51(2H,m,H3′and H5′),7.30(2H,m,H3″and H4′),4.30(2H,dd,J＝7.9,6.6Hz,H6),3.22(2H,ddd,J＝7.9,6.6,2.8Hz,H7),2.40(3H,s,CH ₃ 5″),2.12(3H,s,CH ₃ 4″)。

example 26

Preparation of compound D10:

2.5g of compound C obtained in example 16, namely 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 1.44g of furan-2-formaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound D10, namely (E) -1-phenyl-8- (furan-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 77%, pale yellow solid, melting point: 258-260 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.22(1H,s,H3),8.15(2H,dd,J＝8.7,1.2Hz,H2′and H6′),7.59(1H,br.d,J＝1.8Hz,H5″),7.56(1H,t,J＝2.8Hz,H6″),7.53(2H,dd,J＝8.6,7.4Hz,H3′and H5′),7.36(1H,tt,J＝7.4,1.2Hz,H4′),6.68(1H,d,J＝3.5Hz,H3″),6.56(1H,dd,J＝3.5,1.8Hz,4″),4.26(2H,dd,J＝7.9,6.7Hz,H6),3.35(2H,ddd,J＝7.9,6.7,2.8Hz,H7)。

Example 27

Preparation of compound D11:

compound C obtained in example 16 was 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one 2.5g,10mmol was dissolved in a 20mL mixed solution of absolute ethanol and sodium hydroxide 1.2g,30mmol, then 5-methylfuran-2-carbaldehyde 1.65g,15mmol was slowly added dropwise, the reaction was refluxed until all the starting materials disappeared, the reaction solution was filtered, concentrated, and eluted using a forward silica gel column chromatography gradient with pure ethyl acetate as eluent to give compound D11 as (E) -1-phenyl-8- (5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, yield: 80%, pale yellow solid, melting point: 278-280 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.16(2H,dd,J＝8.6,1.1Hz,H2′and H6′),7.54(2H,dd,J＝8.5,7.5Hz,H3′and H5′),7.52(1H,t,J＝2.7Hz,H6″),7.36(1H,tt,J＝7.5,1.1Hz,H4′),6.60(1H,d,J＝3.3Hz,H3″),6.17(1H,dd,J＝3.3,0.9Hz,H4″),4.27(2H,dd,J＝7.6,7.0Hz,H6),3.34(2H,ddd,J＝7.6,7.0,2.7Hz,H7),2.40(3H,s,CH ₃ 5″)。

EXAMPLE 28

Preparation of compound D12:

2.5g of compound C obtained in example 16 is 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyr-ro-idin-4 (6H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 4, 5-dimethylfuran-2-carbaldehyde 1.86g and 15mmoL are slowly added dropwise, the mixture is refluxed until all the raw materials disappear, the reaction solution is filtered and concentrated, and the mixture is eluted by a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate to obtain the compound D12 which is (E) -1-phenyl-8- (4, 5-dimethylfuran-2-yl) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, the yield is as follows: 77, orange solid, melting point: 254-256 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.21(1H,s,H3),8.16(2H,dd,J＝8.5,1.2Hz,H2′and H6′),7.53(2H,dd,J＝8.5,7.5Hz,H3′and H5′),7.46(1H,t,J＝2.7Hz,H6″),7.35(1H,tt,J＝7.5,1.2Hz,H4′),6.48(1H,s,H3″),4.25(2H,dd,J＝7.7,7.0Hz,H6),3.29(2H,ddd,J＝7.7,7.0,2.7Hz,H7),2.30(3H,s,CH ₃ 5″),2.00(3H,s,CH ₃ 4″)。

Example 29

Preparation of compound D13:

2.5g of compound C obtained in example 16, namely 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, 10mmoL, is dissolved in 20mL of a mixed solution of absolute ethanol and sodium hydroxide of 1.2g,30mmoL, then 2, 3-dihydrobenzo [ D ] [1,4] dioxin-6-carbaldehyde of 2.46g,15mmoL is slowly added dropwise, the mixture is refluxed until all the raw materials disappear, the reaction solution is filtered, concentrated, and the mixture is eluted by adopting a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate, so that the compound D13 is (E) -1-phenyl-8- (2, 3-dihydrobenzo [ b ] [1,4] dioxin-6-ylmethylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is obtained: 90%, pale yellow solid, melting point: 250-252 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.16(2H,dd,J＝8.7,1.1Hz,H2′and H6′),7.68(1H,t,J＝2.8Hz,H7″),7.54(2H,dd,J＝8.4,7.6Hz,H3′and H5′),7.36(1H,tt,J＝7.4,1.1Hz,H4′),7.08(1H,d,J＝2.0Hz,H2″),7.06(1H,dd,J＝8.3,2.0Hz,H6″),6.92(1H,d,J＝8.3Hz,H5″),4.30(4H,m,2xOCH ₂ dioxane),4.26(2H,dd,J＝8.3,6.7Hz,H6),3.29(2H,ddd,J＝8.0,6.7,2.8Hz,H7)。

example 30

Preparation of compound D14:

2.5g of compound C obtained in example 16, namely 1-phenyl-7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, 3.36g of ferrocene formaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate, so that the compound D14 is (E) -1-phenyl-8- (ferrocene-2-methylene) -7, 8-dihydro-1H-pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidin-4 (6H) -one, and the yield is as follows: 89%, orange solid, melting point: 283-285 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.23(1H,s,H3),8.18(2H,d,J＝7.7Hz,H2′and H6′),7.61(1H,t,J＝2.5Hz,H6″),7.57(2H,dd,J＝8.3,7.6Hz,H3′and H5′),7.38(1H,t,J＝7.7Hz,H4′),4.58(2H,J＝1.7Hz,H2″and H5″),4.49(2H,J＝1.7Hz,H3″and H4″),4.26(2H,dd,J＝7.3,7.6Hz,H6),4.19(5H,br.s,H1″′-H5″′),3.06(2H,ddd,J＝7.6,7.6,2.5Hz,H7)。

Example 31

Preparation of Compound E:

2.3g of compound 1-methyl-5 amino-4 pyrazol ethyl formate and 10mmoL are dissolved in 20mL of anhydrous dioxane, 1.5g of valerolactam and 15mmoL are added, 3.8g of phosphorus oxychloride and 25mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction liquid is filtered, concentration is carried out, forward silica gel column chromatography gradient elution is adopted, the eluent is petroleum ether and ethyl acetate in a volume ratio of 1:2, and the compound E is 1-methyl-6, 7,8, 9-tetrahydropyrazole [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -ketone, and the yield is: 88%, white solid, melting point: 166-167 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),4.05(2H,dd,J＝6.7,5.8Hz,H6),3.96(3H,s,NCH ₃ ),2.98(2H,t,J＝6.7Hz,H9),1.96(4H,m,H7,H8)。

example 32

Preparation of compound F1:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazole [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, 1.6g of benzaldehyde and 15mmoL are slowly added dropwise, the mixture is refluxed until all raw materials disappear, the reaction solution is filtered, concentrated, and the mixture is eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate, so that the compound F1 is (E) -1-methyl-9-benzylidene-6, 7,8, 9-tetrahydropyrazole [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is as follows: 82%, white solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.22(1H,t,J＝2.2Hz,H7′),8.07(1H,s,H3),7.46(4H,m,H2′,H3′,H5′and H6′),7.37(1H,m,H4′),4.14(2H,m,H6),4.04(3H,s,NCH ₃ ),2.95(2H,td,J＝6.7,2.1Hz,H8),2.01(2H,m,H7)。

Example 33

Preparation of compound F2:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 1.8g of 3-methylbenzaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate, so that the compound F2 is (E) -1-methyl-9- (2-methylbenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is: 73%, pale yellow solid, melting point: 213-215 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.20(1H,t,J＝2.2Hz,H7′),8.07(1H,s,H3),7.42(2H,d,J＝8.1Hz,H3′and H6′),7.21(2H,d,J＝8.1Hz,H3′and H5′),4.17(2H,m,H6),4.05(3H,s,NCH ₃ ),2.90(2H,td,J＝6.6,2.2Hz,H8),2.01(2H,m,H7).2.37(3H,s,CH ₃ 3″)。

example 34

Preparation of compound F3:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 1.8g of 4-methylbenzaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate, so that the compound F3 is (E) -1-methyl-9- (4-methylbenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is: 84%, pale yellow solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.18(1H,t,J＝2.2Hz,H7′),8.06(1H,s,H3),7.39(2H,d,J＝8.1Hz,H2′and H6′),7.25(2H,d,J＝8.1Hz,H3′and H5′),4.13(2H,m,H6),4.03(3H,s,NCH ₃ ),2.94(2H,td,J＝6.6,2.2Hz,H8),2.40(3H,s,CH ₃ 4″).2.00(2H,m,H7)。

Example 35

Preparation of compound F4:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, 2.1g of 3-chlorobenzaldehyde and 15mmoL are slowly added dropwise, the reaction solution is refluxed until all raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain compound F4 which is (E) -1-methyl-9- (3-chlorobenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is as follows: 69%, pale yellow solid, melting point: 213-215 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.05(1H,s,H3),7.70(1H,t,J＝2.9Hz,H7′),7.51(1H,br.s,H2′),7.42(1H,dt,J＝7.3,1.7Hz,H4′),7.36(1H,dd,J＝7.6,7.3Hz,H5′),7.31(1H,dt,J＝7.6,1.8Hz,H6′),4.14(2H,m,H6),4.01(3H,s,NCH ₃ ),3.37(2H,ddd,J＝7.4,6.9,2.9Hz,H8).2.09(2H,m,H7)。

example 36

Preparation of compound F5:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 1.86g of 4-fluorobenzaldehyde and 15mmoL are slowly added dropwise, the reaction solution is refluxed until all the raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain compound F5 which is (E) -1-methyl-9- (4-fluorobenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is as follows: 52%, pale yellow solid, melting point: 223-225 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.00(1H,s,H3),7.41(2H,d,J＝8.8Hz,H2′and H6′),6.92(1H,s,H7′),6.90(2H,d,J＝8.8Hz,H3′and H5′),4.15(2H,m,H6),4.08(3H,s,NCH ₃ ),2.81(2H,td,J＝7.3,2.1Hz,H8),2.11(2H,m,H7)。

Example 37

Preparation of compound F6:

2.0g of the compound E obtained in example 31, namely 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 2.0g of 4-methoxybenzaldehyde and 15mmoL are slowly added dropwise, the mixture is refluxed until all raw materials disappear, the reaction solution is filtered, concentrated and eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain the compound F6, namely (E) -1-methyl-9- (4-methoxybenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is as follows: 89%, pale yellow solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.47(2H,d,J＝8.8Hz,H2′and H6′),6.98(1H,s,H7′),6.91(2H,d,J＝8.8Hz,H3′and H5′),4.13(2H,m,H6),4.02(3H,s,NCH ₃ ),3.84(3H,s,OCH ₃ 4′),2.86(2H,td,J＝7.3,2.1Hz,H8),2.03(2H,m,H7)。

example 38

Preparation of compound F7:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 2.9g of 3,4, 5-trimethoxybenzaldehyde and 15mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered and concentrated, and forward silica gel column chromatography gradient elution is adopted, and an eluent is pure ethyl acetate, so that the compound F7 is (E) -1-methyl-9- (3, 4, 5-trimethoxybenzylidene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is obtained, and the yield is as follows: 91%, pale yellow solid, melting point: 264-266 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.15(1H,s,H7′),8.07(1H,s,H3),6.71(2H,s,H2′and H6′),4.14(2H,m,H6),4.04(3H,s,OCH ₃ 4′),3.93(OCH ₃ 3′,5′and NCH ₃ ),2.97(2H,td,J＝6.6,2.1Hz,H8),2.03(2H,m,H7)。

Example 38

Preparation of compound F8:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazol [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 1.68g of thiophene-2-formaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate, so that the compound F8 is (E) -1-methyl-9- (thiophen-2-yl) methylene) -6,7,8, 9-tetrahydropyrazol [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is obtained: 69%, pale yellow solid, melting point: 203-205 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.40(1H,t,J＝2.4Hz,H6′),8.06(1H,s,H3),7.57(1H,d,J＝5.1Hz,H5′),7.41(1H,d,J＝3.7Hz,H3′),7.20(1H,dd,J＝5.1,3.7Hz,H4′),4.17(2H,m,H6),4.04(3H,s,NCH ₃ ),2.95(2H,td,J＝6.6,2.3Hz,H8),2.09(2H,m,H7)。

example 40

Preparation of compound F9:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 1.89g of 5-methylthiophene-2-formaldehyde and 15mmoL are slowly added dropwise, the reaction solution is filtered and concentrated until all raw materials disappear, and the reaction solution is eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain compound F9 which is (E) -1-methyl-9- (4, 5-dimethylthiophen-2-yl) methylene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, the yield: 68%, pale yellow solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8,26(1H,t,J＝2.3Hz,H6′),8.04(1H,s,H3),7.11(1H,s,H3′),4.15(2H,m,H6),4.02(3H,s,NCH ₃ ),2.88(2H,m,H8),2.43(3H,s,CH ₃ 5′),2.18(3H,s,CH ₃ 4′),2.05(2H,m,H7)。

Example 41

Preparation of compound F10:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, 10mmoL is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, then 1.44g of furan-2-formaldehyde and 15mmoL are slowly added dropwise, the reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, and the eluent is pure ethyl acetate, so that the compound F10 is (E) -1-methyl-9- (furan-2-yl) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is: 74%, pale yellow solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.05(1H,s,H3),8.01(1H,t,J＝2.3Hz,H6′),7.60(1H,d,J＝1.8Hz,H5′),6.69(1H,d,J＝3.5Hz,H3′),6.57(1H,dd,J＝3.5,1.8Hz,H4′),4.15(2H,m,H6),4.02(3H,s,NCH ₃ ),3.03(2H,td,J＝6.7,2.3Hz,H8),2.04(2H,m,H7)。

example 42

Preparation of compound F11:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 1.65g of 5-methylfuran-2-formaldehyde and 15mmoL are slowly added dropwise, the reaction solution is filtered and concentrated until all raw materials disappear, and the reaction solution is eluted by adopting a forward silica gel column chromatography gradient, and an eluent is pure ethyl acetate to obtain compound F11 which is (E) -1-methyl-9- (5-dimethylfuran-2-yl) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is as follows: 83%, melting point of pale yellow solid: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.96(1H,s,H6′),6.60(1H,d,J＝3.3Hz,H3′),6.17(1H,dd,J＝3.3,Hz,H4′),4.14(2H,m,H6),4.01(3H,s,NCH ₃ ),2.98(2H,td,J＝6.7,2.3Hz,H8),2.41(3H,s,CH ₃ 5′),2.04(2H,m,H7)。

EXAMPLE 43

Preparation of compound F12:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 1.86g of 4, 5-dimethylfuran-2-formaldehyde and 15mmoL are slowly added dropwise, reflux reaction is carried out until all raw materials disappear, the reaction solution is filtered and concentrated, forward silica gel column chromatography gradient elution is adopted, and an eluent is pure ethyl acetate, so that the compound F12 is (E) -1-methyl-9- (4, 5-dimethylfuran-2-yl) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is obtained: 80%, pale yellow solid, melting point: 233-235 DEG C

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.03(1H,s,H3),7.92(1H,t,J＝2.2Hz,H6′),6.50(1H,s,H3′),4.14(2H,m,H6),4.01(3H,s,NCH ₃ ),2.95(2H,td,J＝6.8,2.2Hz,H8),2.31(3H,s,CH ₃ 5′),2.02(2H,m,H7),2.01(3H,s,CH ₃ 4′)。

Example 44

Preparation of compound F13:

2.0g of the compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one and 10mmoL are dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide and 30mmoL, then 2, 3-dihydrobenzo [ F ] [1,4] dioxin-6-formaldehyde 2.46g and 15mmoL are slowly added dropwise, the mixture is refluxed until all the raw materials disappear, the reaction solution is filtered and concentrated, and the mixture is eluted by adopting a forward silica gel column chromatography gradient, wherein an eluent is pure ethyl acetate, so that the compound F13 is (E) -1-methyl-9- (2, 3-dihydrobenzo [ b ] [1,4] dioxin-6-ylmethylene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is obtained: 84%, pale yellow solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.07(1H,t,J＝2.5Hz,H7′),8.04(1H,s,H3),7.02(1H,dd,J＝2.1Hz,H2′),6.99(1H,dd,J＝8.4,2.1Hz,H6′),6.90(1H,d,J＝8.3Hz,H5′),4.29(4H,m,2xOCH ₂ dioxane),4.10(2H,m,H6),4.00(3H,s,NCH ₃ ),2.91(2H,td,J＝6.9,2.2Hz,H8),1.98(2H,m,H7)。

Example 45

Preparation of compound F14:

2.0g of compound E obtained in example 31 is 1-methyl-6, 7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one is dissolved in a mixed solution of 20mL of absolute ethanol and 1.2g of sodium hydroxide, 30mmoL, 3.36g of ferrocene formaldehyde and 15mmoL are slowly added dropwise, the reaction solution is refluxed until all raw materials disappear, the reaction solution is filtered, concentrated, and the elution is carried out by adopting a forward silica gel column chromatography gradient, wherein an eluent is pure ethyl acetate, so that the compound F14 is (E) -1-methyl-9- (ferrocene-2-methylene) -6,7,8, 9-tetrahydropyrazolo [3,4-d ] pyridine [1,2-a ] pyrimidine-4 (1H) -one, and the yield is as follows: 93%, orange solid, melting point: 233-235 ℃;

¹ H NMR(CDCl ₃ with 0.05％v/v TMS,400MHz):δ _H 8.04(1H,s,H3),7.96(1H,br.s,H6′),4.58(2H,br.s,H2′and H5′),4.46(2H,br.s,H3′and H4′),4.20(5H,br.s,H1″-H5″),4.12(2H,m,H6),4.04(3H,s,NCH ₃ ),2.78(2H,td,J＝7.0,1.9Hz,H8),2.01(2H,m,H7)。

example 46

The invention discloses a pyrazolo [3,4-d ] pyrrole (pyridine) o [1,2-a ] pyrimidinone arylene derivative for screening anti-tumor activity:

cell viability was measured by thiazole blue method:

the experimental process comprises the following steps:

selecting cells in logarithmic growth phase, digesting with pancreatin, centrifuging to obtain cell suspension, counting, and mixing each cell with appropriate density (2×10 ⁴ -5×10 ⁴ Per mL), 100 μl per well was inoculated in 96-well plates and incubated in incubator; setting a cell-free zeroing hole, a solvent control hole, a positive medicine control group and a dosing group; after cells are adhered overnight, adding medicine, primarily screening and taking medicine, wherein the final concentration of medicine is 20 mu M, 6 compound holes are arranged for each medicine concentration, continuously incubating for 48 hours after adding medicine, adding 100 mu L of thiazole blue (MTT) for 0.5mg/M for each hole, acting for 4 hours, carefully discarding supernatant, adding 100 mu L of thionyl chloride (DMSO) for each hole, oscillating for 1-3 minutes, and measuring the light absorption value (OD) at 570nm by a multifunctional enzyme-labeled instrument;

The calculation formula is as follows:

percent cell viability = (compound OD-blank OD/control OD-blank OD) ×100%;

cell inhibition ratio% = 1-cell viability% = [1- (compound OD-blank OD/control OD-blank OD)]100% by weight, using graphpad, fitting by formula to obtain IC ₅₀ ；

Sample treatment: dissolving a sample by using thionyl chloride, preserving at low temperature, and controlling the concentration of the thionyl chloride in a final system within a range which does not influence the detection activity;

data processing and result description: under the condition of single concentration of initial screening, for example, monomer compound concentration is 50 mu M, extract is 50 mu g/mu L, the activity of the sample is tested, the inhibition rate is more than 50%, and the dose dependence of the activity, namely IC, is further tested ₅₀ Values obtained by nonlinear fitting of sample concentrations by sample activity, calculated using the software Graphpad Prism 8, and typically, multiple wells (n.gtoreq.3) were set for each sample in the test, and the results were expressed as standard deviations (Standard Deviation, SD) in tables 1-4:

table 1 B1-B14 results of antitumor biological Activity of class-14 derivatives

TABLE 2 results of antitumor biological Activity of D1-D14 derivatives

TABLE 3 results of antitumor biological Activity of F1-F14 derivatives

From the table it can be seen that: the pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives have inhibitory activity on HGC-27 human gastric cancer cells, HT-29 human colon cancer cells, HCT-116 human colon cancer cells, heLa cervical cancer cells, ISK human endometrial cancer cells and MDA-MB-231 human breast cancer cells, and the results show that: the compounds B2, B7, B8, B10, B12, D2-D13, D14 and F1-F13 have inhibitory activity on HGC-27 human gastric cancer cells; compounds B2, B7, B8, B12, B13, D2-D12, D14, F1-F14 have inhibitory activity against HT-29 human colon cancer cells; the compounds B2, B7, D2-D4, F1-F9, F12 and F13 have inhibitory activity on HCT-116 human colon cancer cells; the compounds B1, B2, B4, B7, B10, D2, D3, D5, D7, D10, F1-F4, F6-F9 and F11-F13 have inhibitory activity on HeLa cervical cancer cells; the compounds B1, B2, B4, B7, B10, D2-D5, D7, D10, F1, F2, F4, F6, F7, F9, F11-F13 have inhibitory activity on ISK human endometrial cancer cells; compounds B2, B7, B12, D2-D5, D10, F1-F5, F7, F9, F11-F13 have inhibitory activity against MDA-MB-231 human breast cancer cells.

Claims

1. Pyrazolo [3,4-d ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives or pyrazolo [3,4-d ] pyrido [1,2-a ] pyrimidinone arylene derivatives, characterized in that the structural formula of the derivatives is:

wherein:

2. Use of pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivatives or pyrazolo [3,4-D ] pyrido [1,2-a ] pyrimidinone arylene derivatives of compounds B2, B7, B8, B10, B12, D2-D12, D14, F1-F13 in the manufacture of a medicament for the treatment of HGC-27 human gastric cancer according to claim 1.

3. Use of a pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivative or a compound B2, B7, B8, B12, B13, D2-D12, D14, F1-F14 in pyrazolo [3,4-D ] pyrido [1,2-a ] pyrimidinone arylene derivative according to claim 1 for the manufacture of a medicament for the treatment of HT-29 human colon cancer.

4. Use of a pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivative or a compound B2, B7, D2-D4, F1-F9, F12, F13 in pyrazolo [3,4-D ] pyrido [1,2-a ] pyrimidinone arylene derivative according to claim 1 for the preparation of a medicament for the treatment of HCT-116 human colon cancer.

5. Use of a pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivative or a compound B1, B2, B4, B7, B10, D2, D3, D5, D7, D10, F1-F4, F6-F9, F11-F13 in pyrazolo [3,4-D ] pyrido [1,2-a ] pyrimidinone arylene derivative according to claim 1 for the manufacture of a medicament for the treatment of Hela cervical cancer.

6. Use of a pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivative or a pyrazolo [3,4-D ] pyrido [1,2-a ] pyrimidinone arylene derivative of the compound B1, B2, B4, B7, B10, D2-D5, D7, D10, F1, F2, F4, F6, F7, F9, F11-F13 for the preparation of a medicament for the treatment of ISK human endometrial cancer.

7. Use of a pyrazolo [3,4-D ] pyrrolo [1,2-a ] pyrimidinone arylene derivative or a pyrazolo [3,4-D ] pyrido [1,2-a ] pyrimidinone arylene derivative of compound B2, B7, B12, D2-D5, D10, F1-F5, F7, F9, F11-F13 in the manufacture of a medicament for the treatment of MDA-MB-231 human breast cancer.