CN109503586B

CN109503586B - Dihydropteridinone BRD4 protein inhibitor, preparation method and application thereof

Info

Publication number: CN109503586B
Application number: CN201811582864.3A
Authority: CN
Inventors: 刘宏民; 郑一超; 孙凯; 李雯; 符运栋; 宋启梦
Original assignee: Zhengzhou University
Current assignee: Zhengzhou University
Priority date: 2018-12-24
Filing date: 2018-12-24
Publication date: 2021-05-07
Anticipated expiration: 2038-12-24
Also published as: CN109503586A

Abstract

本发明涉及药物化学领域，具体涉及一种新的二氢蝶啶酮类BRD4蛋白抑制剂、它们的制备方法及用途，特别是作为BRD4蛋白抑制剂及抗肿瘤药物方面的用途。其结构通式如下：

The invention relates to the field of medicinal chemistry, in particular to a novel dihydropteridone BRD4 protein inhibitor, a preparation method and application thereof, and in particular to the application as a BRD4 protein inhibitor and an antitumor drug. Its general structure is as follows:

Description

Dihydropteridinone BRD4 protein inhibitor, preparation method and application thereof

Technical Field

The invention relates to the field of pharmaceutical chemistry, in particular to a dihydropteridinone BRD4 protein inhibitor, a preparation method thereof, a medicinal composition containing the compounds and application thereof, especially application of the dihydropteridinone BRD4 protein inhibitor as a BRD4 protein inhibitor and an antitumor drug.

Background

Malignant tumor is the first major disease threatening the health of people in China. 2017, cancer data of 2013 was published by "Chinese cancer research" published by the national cancer center, heijid, et al. The report indicates that cancer is the first cause of death in urban and rural residents in China; the number of new cancer cases continuously rises, and 368 thousands of cancer cases in 2013 account for 1/4 of new cases in the world; about 1 million people per day throughout the country have been diagnosed with cancer. Therefore, the research of the antitumor drugs and the related target inhibitors thereof has important clinical and social values.

Brd (bromodomain), also known as bromodomain protein, is a conserved protein domain that specifically recognizes acetylated lysine residues, and plays a key role in regulation of chromatin assembly and gene transcription. Multiple studies prove that BRD is a highly adaptable drug target, and small-molecule inhibitors acting on BRD have wide prospects in tumor treatment.

BRD4 is a member of the bromodomain protein family, and its biological function is most widely and extensively studied. BRD4 recognizes not only promoter regions, but also intergenic and intragenic regions, which are involved in gene expression. The BRD4 protein performs a transcriptional regulatory function in physiological conditions, regulating normal cellular processes; while in abnormal states, they recruit various proteins to chromatin and transcription sites, regulating the transcription of genes closely related to gene expression, such as c-Myc and BCL-2. BRD4 recruits proteins at the intercellular level in a positive transcription elongation factor (P-TEFb) dependent manner, and regulates the transcription process by coupling with RNA polymerase II, thereby promoting abnormal expression of disease-related genes.

A great deal of research shows that the BRD4 protein has a close relationship with the occurrence and development of tumors, for example, the Bromodomain coding region of BRD4 and NUT (nucleoprotein in testis) gene are ectopic to form BRD-NUT fusion type proto-oncogene, which can cause midline cancer; inhibition of BRD4 expression in melanoma significantly slows the growth of melanoma cells; inhibiting expression of BRD4 in malignant peripheral nerve sheath tumor cells, resulting in death of the tumor cells; inhibition of BRD4 can inhibit proliferation of hepatocarcinoma cells; by interfering with the binding of BRD4 to the oncogene MYC, silencing of the oncogene MYC can be directly caused in models of various hematopoietic tumors, such as AML, Burkitt's lymphoma, multiple myeloma, and B-cell acute lymphocytic leukemia. Bromodomain proteins can regulate the assembly of histone acetylation-dependent chromatin complexes expressed by inflammatory genes and regulate transcription of inflammatory genes. Research has shown that BRD4 can act as a transcriptional co-activator of NF-Kb to promote inflammation. In the field of cardiovascular diseases, BRD4 has also been found to play a key co-activator of pathological gene transactivation during cardiomyocyte hypertrophy, playing an important role in heart failure; the BRD4 small molecule inhibitor JQ1 can be a potential drug for treating heart failure and improve the cardiac function.

The potential value of the bromodomain protein family in anti-inflammatory and anti-tumor aspects has attracted high attention of various pharmaceutical companies, colleges and scientific research institutions all over the world, and BRD4 also becomes an important target in related fields. The BRD4 is taken as a target, and a small molecule inhibitor with high activity and high selectivity is found to become a research hotspot for treating diseases such as tumor, inflammation, cardiovascular disease, AIDS and the like at present, so that the BRD4 has wide development and application prospects.

At present, various BRD4 small molecule inhibitors represented by JQ1 have been reported, and related compounds OTX-015, I-BET762 and TEN-010 have entered clinical stage research. However, the number of BRD4 small-molecule inhibitors currently entering clinical research is few, and there is an urgent need to find more novel and efficient BRD4 inhibitors, which provide new options for treatment of diseases such as tumors.

Disclosure of Invention

The invention aims to provide a novel compound with a dihydropteridinone structure and a preparation method thereof, and also aims to provide an application of the compound as an antitumor drug in the aspect of inhibiting a bromine-containing structural domain BRD 4.

The specific technical scheme is as follows:

the general formula (I) of the novel dihydropteridinone compound is as follows:

R¹is alkyl, aryl, heterocyclic radical;

alkyl is a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms to which a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms is attached, wherein each carbon atom is optionally substituted with oxygen; aryl is selected from phenyl, benzyl, naphthyl; a monocyclic heterocycle selected from the group consisting of piperidinyl, pyrrolyl, pyrazolyl, imidazolyl, furanyl, indolyl, morpholinyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, piperazinyl, substituted piperazinyl, pyrazinyl or pyridazinyl; each aryl and heterocyclyl is optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from the group consisting of halogen, haloalkyl, alkyl, cyano, hydroxy, nitro, mercapto, alkoxy, alkylthio, aralkyl, diarylalkyl;

R²is hydrogen, C1-C3 alkyl;

R³is alkyl, aryl, heterocyclic radical;

the alkyl is a straight chain or branched chain saturated hydrocarbon group containing 1 to 8 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 8 carbon atoms connecting straight chain or branched chain saturated hydrocarbon groups having 1 to 8 carbon atoms, wherein each carbon atom is optionally substituted by oxygen or nitrogen atom; aryl is selected from phenyl, benzyl, naphthyl; heterocyclyl is a monocyclic heterocycle selected from piperidinyl, pyrrolyl, pyrazolyl, imidazolyl, furanyl, morpholinyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, piperazinyl, substituted piperazinyl, pyrazinyl or pyridazinyl; or a bicyclic heterocycle of quinolinyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothienyl, benzisothiazolyl, benzofuranyl, benzodioxanyl, benzodioxolyl; each aryl, monocyclic or bicyclic heterocycle is optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from the group consisting of halogen, haloalkyl, alkyl, cyano, hydroxy, mercapto, amino, nitro, alkoxy, alkylthio, aralkyl, diarylalkyl, or the following aliphatic heterocycles: tetrahydropyrrolyl, morpholinyl, alkoxymorpholinyl, piperazinyl, piperidinyl, alkylaminopiperidinyl;

the preferred scheme of the invention is as follows:

R¹is methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, benzyl, naphthyl, thienyl, furyl, phenylpropenyl, indolyl, pyrrolyl or pyridyl;

R²hydrogen and methyl;

R³is phenyl, methyl monosubstituted phenyl, tert-butyl monosubstituted phenyl, halogen monosubstituted phenyl, substituted byHydroxy, methoxy substituted phenyl, halo disubstituted phenyl, dimethoxy substituted phenyl, hydroxy monosubstituted phenyl, hydroxy, halo substituted phenyl, nitro substituted phenyl, ethoxy, hydroxy substituted phenyl, methoxy monosubstituted phenyl, diethylamino, hydroxy substituted phenyl, acetonitrile monosubstituted phenyl, naphthyl, thienyl, furanyl, phenylpropenyl, indolyl, pyrrolyl, methyl monosubstituted thienyl, hydroxy monosubstituted naphthyl, methoxy monosubstituted furanyl, pyridyl, methyl monosubstituted pyridyl, methoxy monosubstituted pyridyl, amino monosubstituted pyridyl, halo disubstituted pyridyl, nitro monosubstituted pyridyl, halo trisubstituted pyridyl.

The preferable scheme is as follows: r¹Is a cyclic saturated hydrocarbon group consisting of 3-6 carbon atoms;

R²hydrogen and methyl;

R³is phenyl, substituted phenyl, naphthyl, substituted naphthyl, thienyl, substituted thienyl, furyl, substituted furyl, phenylpropenyl, indolyl, substituted indolyl, pyrrolyl, substituted pyrrolyl, pyridyl, substituted pyridyl; the substituent is selected from C1-3 alkyl, halogen, C1-3 alkoxy, hydroxyl, nitro, amino, C1-3 alkyl substituted amino and cyano.

More preferably, the scheme is as follows: r¹Is cyclopentyl;

R²hydrogen and methyl;

R³phenyl, methyl mono-substituted phenyl, halogen mono-or di-substituted phenyl, methoxy mono-or di-substituted phenyl, ethoxy mono-or di-substituted phenyl, cyano mono-substituted phenyl, amino mono-substituted phenyl, phenyl mono-substituted by diethylamino; naphthyl, thienyl, furyl, phenylpropenyl, indolyl, pyrrolyl, methyl monosubstituted thienyl, pyridyl, methyl monosubstituted pyridyl, methoxy monosubstituted pyridyl, amino monosubstituted pyridyl, halogen monosubstituted pyridyl, nitro monosubstituted pyridyl.

Preferred compounds of the general formula (I) according to the invention are those of the following structure:

(E) -2- (2-benzylidenehydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (2-methylbenzylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (3-methylbenzylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- (2-fluorobenzyl) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- (3-fluorobenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- (4-fluorobenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-2- (2- (2, 3-difluorobenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-2- (2- (2, 4-difluorobenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-2- (2- (2, 6-difluorobenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (1-phenylethylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- (2-methoxybenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- (3-methoxybenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-2- (2- (3, 4-dimethoxybenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-2- (2- (3-ethoxy-2-hydroxybenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-2- (2- (4- (diethylamino) -2-hydroxybenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -2- ((2- (8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5, 6,7, 8-tetrahydropteridin-2-yl) hydrazono) methyl) benzonitrile

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (naphthalen-1-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((1E,2E) -3-phenylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((1H-indol-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (thien-2-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((5-methylthiophen-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- (furan-2-ylmethylene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((1H-pyrrol-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (pyridin-2-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((6-chloropyridin-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((5-methylpyridin-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((6-methylpyridin-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((4-chloropyridin-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- ((6-methoxypyridin-2-yl) methylene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((5-chloropyridin-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (pyridin-3-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((2-chloropyridin-3-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -2- (2- ((2-aminopyridin-3-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (1- (pyridin-3-yl) ethylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-2- (2- ((5-methoxypyridin-3-yl) methylene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (pyridin-4-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- (1- (pyridin-4-yl) ethylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((3-nitropyridin-4-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

(E) -8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((4-methylpyridin-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one

The specific structure is shown in table one:

TABLE I preferred Compound Structure

According to the invention, pharmaceutically acceptable salts include the acid addition salts formed with the following acids of general formula (I): hydrochloric, hydrobromic, sulphuric, phosphoric, borate, methanesulphonic, p-toluenesulphonic, naphthalenesulphonic, benzenesulphonic, citric, lactic, pyruvic, tartaric, acetic, maleic or succinic, mandelic, fumaric, salicylic, phenylacetic.

The invention further provides a preparation method of the compound shown in the general formula (I).

The preparation method of the compound with the general formula (I) comprises the following steps:

a. dissolving the compound 1 in methanol, dropwise adding thionyl chloride in an ice bath, carrying out reflux reaction, after the reaction is finished, carrying out reduced pressure concentration on a reaction system, separating out a product, and filtering to obtain a product 2, wherein the crude product is subjected to the next reaction without purification. The yield was about 90%.

b. Dissolving the intermediate 2 in a solvent, adding a corresponding carbonyl compound, adding a proper amount of an alkaline compound and a reducing agent under ice bath, and stirring at room temperature until the raw materials disappear. Quenching reaction after the reaction is finished, extracting, combining organic phases, drying, filtering and concentrating to obtain an intermediate 3 crude product, and directly carrying out the next reaction without purification. The yield was 70%. The solvent can be chloroform, dichloromethane, dichloroethane; the alkaline compound may be Na₂CO₃、K₂CO₃、CsCO₃NaOAc, KOAc; the reducing agent used may be NaBH₄、KBH₄、NaBH(OAc)₃、NaBH₃CN。

c. Dissolving the intermediate 3 in a solvent, adding 2, 4-dichloro-5-nitropyrimidine and a corresponding alkaline compound under ice bath, and stirring at room temperature until the raw materials disappear. After the reaction is finished, the solvent is removed by decompression and concentration, and the intermediate 4 is obtained by silica gel column chromatography separation after dilution, washing and organic layer drying. The yield was 85%. The solvent can be chloroform, dichloromethane, dichloroethane; the alkaline compound may be Na₂CO₃、K₂CO₃、CsCO₃、NaOAc、KOAc。

d. Dissolving the intermediate 4 in acetic acid, heating to 50-70 ℃, carrying out batch reduction on iron powder, stirring for reaction, and heating to 100-110 ℃ for continuous reaction. After the reaction is finished, filtering, washing residues, concentrating the filtrate, and performing column chromatography to separate the intermediate 5. The yield was about 60%.

e. Intermediate 5 was added to anhydrous DMF and iodomethane (1.3 eq) and NaH (1.3 eq) were added under ice bath. Stirring and reacting at room temperature, quenching the reaction system after the reaction is finished, extracting, washing the organic phase with water, drying and concentrating to obtain an intermediate 6, and the intermediate can be directly used for the next reaction without purification. The yield was 90%.

f. Dissolving the intermediate 6 in ethanol, adding hydrazine hydrate solution (2 equivalents), sealing and reacting at 80-90 ℃. After the reaction is finished, the reaction system is concentrated, and the intermediate 7 is obtained after washing, drying and concentrating, and the product can be used for the next reaction without purification. The yield was 87%.

g. Dissolving the intermediate 7 and corresponding aldehyde or ketone in absolute ethyl alcohol, and stirring at 50-60 ℃ until the raw materials are not reduced. And after the reaction is finished, decompressing and concentrating to remove the solvent, washing with water, drying, and purifying by concentration column chromatography to obtain the target product 8. The yield was 80%.

The invention provides application of a compound with a general formula (I) in BRD4 protein inhibition and antitumor activity.

The in vitro BRD4 protein inhibitory activity was tested on a portion of the compounds of formula (I) and the results are shown in Table II.

TABLE II protein inhibitory Activity of the partial Compound BRD4 of the present invention

The compounds of partial general formula (I) were tested for their inhibitory activity against tumor cells in vitro, and the results are shown in Table III.

TABLE III partial compound of the invention has in vitro antitumor activity

Note: JQ1 is a positive control; SGC, BGC, MGC, MKN45 are gastric cancer cells, THP1 is leukemia cell

The biological activity test result shows that the compound of the general formula (I) provided by the invention has good BRD4 protein inhibition effect and good tumor cell inhibition effect. The inhibitory activity of partial compounds on gastric cancer cells reaches or is better than that of a positive control compound JQ 1.

Accordingly, the compounds of the present invention are useful as inhibitors of BRD4 in the treatment of various BRD4 mediated diseases. The compound related to the invention can be used for preventing or treating various parenchymal organ tumors related to BRD4, including but not limited to gastric cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, prostate cancer, leukemia, lymph cancer and the like.

The invention has the advantages that a reaction route is designed, the target compound is obtained through seven steps of reaction, and the total yield can reach 20-25%. Experiments prove that the novel compound with the dihydropteridinone structure has good BRD4 protein inhibition effect and good tumor cell inhibition effect, and especially aiming at gastric cancer cells, the inhibition activity of part of compounds reaches or is superior to that of a positive control compound JQ 1. Provides a new drug choice for treating BRD4 inhibitor, tumor and other diseases.

Detailed Description

The invention is further described below with reference to examples, which are intended to be illustrative only and not to limit the scope of the invention.

The structure of the compounds was determined by Nuclear Magnetic Resonance (NMR) and High Resolution Mass Spectrometry (HRMS). The nuclear magnetic resonance apparatus is a Bruker DPX-400 type superconducting nuclear magnetic resonance apparatus in Sweden, and Tetramethylsilane (TMS) is used as an internal standard; the high resolution mass spectrometer used was a Waters-Micromass Q-Tof mass spectrometer.

Example 1

Preparation of methyl 2-aminobutyric acid hydrochloride:

2-aminobutyric acid (1.00g, 1.0eq.) was put in a 50mL eggplant-shaped bottle, anhydrous methanol (15 mL) was added, and SOCl was slowly added at 0 deg.C₂(2.31g, 2.0eq.) TLC showed the reaction was complete after heating to reflux for 1.5 hours. Evaporation of the volatiles gave a white solid, which was washed with Et₂And O, grinding the solid, and performing suction filtration to obtain a crude product of the compound 2, wherein the crude product can be directly subjected to the next reaction without purification, and the yield is 90%.

White solid, melting point 120-122 ℃.¹H NMR(400MHz,D₂O)δ4.04(t,J＝6.2Hz,1H),3.76(s,3H),1.99–1.82(m,2H),0.97–0.87(m,3H).¹³C NMR(101MHz,D₂O)δ170.79,53.99,53.46,23.21,8.42.HR-MS(ESI):Calcd.C₅H₁₂ClNO₂,[M+H]⁺m/z:118.0863,found:118.0867.

Example 2

Preparation of methyl 2- (cyclopentylamino) butyrate:

compound 2(1.00g, 1.0eq.) and cyclopentanone (548mg, 1.0eq.) were ice-cooled for 10 minutes, and NaOAc (547mg, 1.0eq.) and NaBH (OAc) were added₃(2.07g, 1.5 eq.). The reaction mixture was stirred at room temperature overnight. TLC shows that after the reaction is finished, saturated NaHCO is added into the reaction system₃The reaction was quenched in solution and then with CH₂Cl₂(20 mL. times.2) the aqueous layer was extracted, the organic layers were combined and washed with anhydrous Na₂SO₄Drying, filtration and concentration gave compound 3 in 65% yield.

A colorless oily liquid.¹H NMR(400MHz,DMSO-d⁶)δ3.63(s,3H),3.10(t,J＝6.6Hz,1H),2.98–

2.84(m,1H),1.80(s,1H),1.67–1.54(m,4H),1.54–1.48(m,2H),1.47–1.39(m,2H),1.32–1.21(m,2H),0.84(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ175.71,60.88,57.32,51.19,33.17,32.10,26.25,23.38,23.33,10.21.HR-MS(ESI):Calcd.C₁₀H₁₉NO₂,[M+H]⁺m/z:186.1494,found:186.1495.

Example 3

Preparation of methyl 2- ((2-chloro-5-nitropyrimidin-4-yl) (cyclopentyl) amino) butanoate:

adding compound 3(1.00g, 1.0eq.) into acetone 15mL, adding 2, 4-dichloro-5-nitropyrimidine (1.05g, 1.0eq.) after ice bath for 10 minutes, and then adding K₂CO₃(750mg, 1.0eq.), and stirred at room temperature overnight. After completion of the reaction, the solvent was removed, the residue was dissolved with ethyl acetate and washed with water, and the organic layer was collected, dried over anhydrous sodium sulfate, and purified by silica gel flash column chromatography (PE/EA ═ 10:1) to obtain compound 4 in 89% yield.

Yellow solid, melting point 110-111 ℃.¹H NMR(400MHz,DMSO-d⁶)δ8.84(s,1H),4.29–4.14(m,1H),3.64(s,3H),3.52–3.48(m,1H),2.28–2.15(m,1H),2.04–1.90(m,2H),1.85–1.77(m,2H),1.72–1.61(m,3H),1.51–1.35(m,2H),0.94(t,J＝7.8Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ164.21,151.73,151.14,137.83,119.27,61.06,59.38,28.30,28.28,26.36,23.88,23.86,8.33.HR-MS(ESI):Calcd.C₁₄H₁₉ClN₄O₄,[M+Na]⁺m/z:365.0993,found:365.0992.

Example 4

Preparation of 2-chloro-8-cyclopentyl-7-ethyl-7, 8-dihydropteridin-6 (5H) -one:

compound 4(1.00g, 1.0eq.) was added to 20mL of glacial acetic acid, heated to 70 ℃, and iron powder (325mg, 2.0eq.) was added in portions over 5 minutes. The reaction was stirred at 70 ℃ for 1 hour and then warmed to 100 ℃ for 4-5 hours, after completion of the reaction, the mixture was filtered through celite, the residue was washed with appropriate methanol, the volatiles were evaporated and purified by flash column chromatography (PE/EA ═ 1.5:1) to give compound 5 in 50% yield.

Yellow solid, melting point 180-182 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.85(s,1H),7.57(s,1H),4.30–4.20(m,1H),4.18–4.04(m,1H),2.00–1.76(m,7H),1.76–1.65(m,1H),1.62–1.42(m,2H),0.92–0.68(m,3H).¹³C NMR(101MHz,DMSO-d⁶)δ164.21,151.73,151.14,137.83,119.27,61.06,59.38,28.30,28.28,26.36,23.88,23.86,8.33.HR-MS(ESI):Calcd.C₁₃H₁₇ClN₄O,[M+Na]⁺m/z:303.0989,found:303.0975.

Example 5

Preparation of 2-chloro-8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

adding the compound 5(1.00g, 1.0eq.) into anhydrous DMF, carrying out ice bath for 10 minutes, adding iodomethane (657mg, 1.3eq.) and NaH (185mg, 1.3eq.) with the mass percent of 60% for stirring at room temperature for 3 hours, after the reaction is finished, quenching the reaction system by using ice water, extracting the product by using EtOAc, washing an organic layer twice by using water, drying by using anhydrous sodium sulfate, and carrying out spin drying to obtain the compound 6, wherein the compound 6 can be directly used in the next step without purification and has the yield of 90%.

Yellow solid, melting point 182 ℃ and 183 ℃.¹H NMR(400MHz,DMSO-d⁶)δ7.87(s,1H),4.39–4.31(m,1H),4.17(p,J＝8.3Hz,1H),3.24(s,3H),1.97–1.77(m,7H),1.74–1.65(m,1H),1.61–1.48(m,2H),0.74(t,J＝7.5Hz,3H).¹³C NMR(101MHz,DMSO)δ163.16,152.13,151.53,138.49,121.24,60.76,59.32,28.38,28.33,27.91,26.61,23.85,23.81,8.48.HR-MS(ESI):

Calcd.C₁₄H₁₉ClN₄O,[M+H]⁺m/z:295.1325,found:295.1322.

Example 6

Preparation of 8-cyclopentyl-7-ethyl-2-hydrazino-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

dissolving a compound 6(1.00g, 1.0eq.) in ethanol, adding hydrazine hydrate (678mg, 2eq.) with the mass percentage content of 50%, reacting in a water-heating reaction kettle at 90 ℃ for 48 hours, evaporating off volatile matters after the reaction is finished, dissolving the volatile matters in ethyl acetate, washing by adding water, drying an organic phase, and performing spin drying to obtain a compound 7, wherein the compound 7 is directly put into the next step without purification, and the yield is 70%.

White solid, melting point 120-121 ℃.¹H NMR(400MHz,DMSO-d⁶)δ7.68(s,1H),7.42(s,1H),4.35–4.21(m,1H),4.17–4.10(m,1H),4.04(s,2H),1.97–1.68(m,7H),1.64–1.47(m,3H),0.75(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.80,160.49,151.56,138.60,114.30,60.21,58.19,28.73,28.61,27.68,26.17,23.48,23.31,8.90.HR-MS(ESI):Calcd.C₁₄H₂₂N₆O,[M+H]⁺

m/z:291.1933,found:291.1934.

Example 7

(E) Preparation of-2- (2-benzylidenehydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

compound 7(100mg, 1.0eq.) and benzaldehyde (40mg, 1.1eq.) were added to 8mL of absolute ethanol, reacted at 60 ℃ for 8 hours, detected by TLC (1:2PE/EA), and after completion of the reaction, purified by column chromatography (1:1PE/EA) to give compound 8 in 69% yield. White solid, melting point 162-.¹H NMR(400MHz,DMSO-d⁶)δ10.73(s,1H),8.06(s,1H),7.76(s,1H),7.63(d,J＝7.3Hz,2H),7.39(t,J＝7.4Hz,2H),7.31(t,J＝7.3Hz,1H),4.26–4.19(m,1H),4.16–4.05(m,1H),3.23(s,3H),2.18–2.08(m,1H),2.08–1.85(m,5H),1.85–1.75(m,1H),1.73–1.63(m,1H),1.62–1.47(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.93,155.21,151.36,139.31,138.52,135.72,128.58,128.35,125.89,115.62,61.57,59.54,28.67,28.55,27.64,26.25,23.96,23.86,8.70.HR-MS(ESI):Calcd.C₂₁H₂₆N₆O,[M+H]⁺m/z:379.2446,found:379.2447.

Example 8

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (2-methylbenzylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the same procedure used in example 7 was repeated except for using 2-methylbenzaldehyde instead of benzaldehyde, whereby the yield was found to be 71%.

White solid, melting point 231-.¹H NMR(400MHz,DMSO-d⁶)δ10.66(s,1H),8.33(s,1H),7.81–7.75(m,2H),7.21(t,J＝4.3Hz,3H),4.24–4.20(m,1H),4.17–4.09(m,1H),3.23(s,3H),2.42(s,3H),2.08(d,J＝11.2Hz,1H),2.00–1.89(m,5H),1.82–1.75(m,1H),1.71–1.64(m,1H),1.60–1.49(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.92,155.25,151.44,138.57,138.20,135.35,133.55,130.68,128.04,125.83,125.16,115.57,61.30,59.34,28.58,28.53,27.65,26.26,23.72,23.65,19.20,8.72.HR-MS(ESI):Calcd.C₂₂H₂₈N₆O,[M+H]⁺m/z:393.2403,found:393.2404.

Example 9

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (3-methylbenzylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was conducted using 3-methylbenzaldehyde instead of benzaldehyde, and the yield was 70%.

White solid, melting point 220-222 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.84(s,1H),8.03(s,1H),7.72(s,1H),7.52(s,1H),7.40(d,J＝7.7Hz,1H),7.28(t,J＝7.6Hz,1H),7.14(d,J＝7.4Hz,1H),4.27–4.23(m,1H),4.14–4.05(m,1H),3.23(s,3H),2.33(s,3H),2.19–2.10(m,1H),2.07–1.89(m,5H),1.84–1.78(m,1H),1.73–1.66(m,1H),1.61–1.50(m,1H),0.76(t,J＝7.4Hz,2H).¹³C NMR(101MHz,DMSO-d⁶)δ162.90,154.58,151.45,140.09,137.69,135.45,129.27,128.49,126.19,123.67,115.66,61.67,59.78,28.60,28.47,27.69,26.32,23.98,23.85,20.91,8.62.HR-MS(ESI):Calcd.C₂₂H₂₈N₆O,[M+H]⁺m/z:393.2403,found:393.2402.

Example 10

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- (2-fluorobenzyl) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the procedure of example 7 was followed, using 2-fluorobenzaldehyde instead of benzaldehyde, in a yield of 70%.

White solid, melting point 162-.¹H NMR(400MHz,DMSO-d⁶)δ10.92(s,1H),8.28(s,1H),7.95–7.83(m,1H),7.77(s,1H),7.45–7.32(m,1H),7.30–7.20(m,2H),4.28–4.19(m,1H),4.16–4.05(m,1H),3.24(s,3H),2.16–2.08(m,1H),2.06–1.89(m,5H),1.84–1.76(m,1H),1.73–1.64(m,1H),1.61–1.49(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.95,161.22,158.75,155.00,151.36,138.46,131.64,131.59,130.02,129.94,115.86,115.67,61.58,59.59,28.63,28.50,27.65,26.27,23.95,23.83,8.67.HR-MS(ESI):Calcd.C₂₁H₂₅FN₆O,[M+H]⁺m/z:397.2152,found:397.2153.

Example 11

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- (3-fluorobenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was conducted using 3-fluorobenzaldehyde instead of benzaldehyde, and the yield was 74%.

White solid, melting point 181-183 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.88(s,1H),8.04(s,1H),7.76(s,1H),7.51–7.37(m,3H),7.18–7.08(m,1H),4.27–4.22(m,1H),4.11–4.02(m,1H),3.24(s,3H),2.20–2.12(m,1H),2.08–1.88(m,5H),1.83–1.76(m,1H),1.72–1.65(m,1H),1.61–1.50(m,2H),0.76(t,J＝7.5Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.69,162.97,161.27,155.03,151.34,138.47,138.42,137.70,130.65,130.57,122.51,115.81,115.08,114.87,111.42,111.20,61.78,59.79,28.60,28.44,27.63,26.27,24.00,23.82,8.64.HR-MS(ESI):Calcd.C₂₁H₂₅N₆O,[M+H]⁺m/z:397.2152,found:397.2151.

Example 12

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- (4-fluorobenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was conducted using 4-fluorobenzaldehyde instead of benzaldehyde, and the yield was 74%.

White solid, melting point 210-212 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.72(s,1H),8.05(s,1H),7.75(s,1H),7.71–7.62(m,2H),7.23(t,J＝8.9Hz,2H),4.24–4.20(m,1H),4.17–4.08(m,1H),3.24(d,J＝4.4Hz,3H),2.15–1.88(m,6H),1.83–1.75(m,1H),1.72–1.63(m,1H),1.61–1.47(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.32,162.94,160.88,155.19,151.38,138.50,138.25,132.31,132.28,127.82,127.74,115.69,115.64,115.47,61.47,59.45,28.67,28.56,27.65,26.26,23.93,23.84,8.70.HR-MS(ESI):Calcd.C₂₁H₂₅FN₆O,[M+H]⁺m/z:397.2154,found:397.2155.

Example 13

(E) Preparation of-8-cyclopentyl-2- (2- (2, 3-difluorobenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the procedure of example 7 was repeated using 2, 3-difluorobenzaldehyde instead of benzaldehyde, whereby the yield was 73%.

White solid, melting point:151-152℃.¹H NMR(400MHz,DMSO-d⁶)δ11.04(s,1H),8.26(s,1H),7.78(s,1H),7.69(t,J＝7.0Hz,1H),7.42–7.31(m,1H),7.27–7.18(m,1H),4.28–4.21(m,1H),4.16–4.04(m,1H),3.24(s,3H),2.17–1.88(m,6H),1.84–1.76(m,1H),1.72–1.64(m,1H),1.62–1.50(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.96,154.81,151.36,138.39,130.46,125.72,125.65,124.72,120.46,116.58,116.41,116.07,61.54,59.59,28.59,28.47,27.66,26.29,23.92,23.80,8.66.HR-MS(ESI):Calcd.C₂₁H₂₄F₂N₆O,[M+H]⁺m/z:415.2058,found:415.2058.

example 14

(E) Preparation of-8-cyclopentyl-2- (2- (2, 4-difluorobenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the method is the same as example 7 except for using 2, 4-difluorobenzaldehyde instead of benzaldehyde, and the yield is 80 percent

White solid, melting point 181-182 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.91(s,1H),8.22(s,1H),7.95–7.88(m,1H),7.76(s,1H),7.34–7.26(m,1H),7.16(t,J＝8.5Hz,1H),4.27–4.20(m,1H),4.15–4.07(m,1H),3.23(s,3H),2.13–1.88(m,6H),1.82–1.75(m,1H),1.71–1.63(m,1H),1.55(d,J＝13.5Hz,2H),0.76(t,J＝7.5Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.94,154.98,151.36,138.46,130.92,126.54,120.11,115.88,112.35,112.13,104.46,104.21,61.50,59.50,28.64,28.53,27.65,26.27,23.93,23.83,8.68.HR-MS(ESI):Calcd.C₂₁H₂₄F₂N₆O,[M+H]⁺m/z:

415.2058,found:415.2059.

Example 15

(E) Preparation of-8-cyclopentyl-2- (2- (2, 6-difluorobenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the procedure of example 7 was repeated except for using 2, 6-difluorobenzaldehyde instead of benzaldehyde, whereby the yield was 80%.

White solid, melting point 238-.¹H NMR(400MHz,DMSO-d⁶)δ10.91(s,1H),8.19(s,1H),7.77(s,1H),7.44–7.33(m,1H),7.19–7.08(m,2H),4.25–4.19(m,1H),4.19–4.09(m,1H),3.24(s,3H),2.03–1.84(m,6H),1.82–1.75(m,1H),1.69–1.62(m,1H),1.50(t,J＝12.9Hz,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.96,161.18,161.11,158.67,158.60,154.90,151.53,138.48,129.81,129.70,129.17,115.99,112.91,112.77,112.63,112.15,112.09,111.90,60.90,59.11,28.53,28.50,27.67,26.26,23.40,8.75.HR-MS(ESI):Calcd.C₂₁H₂₄F₂N₆O,[M+H]⁺m/z:415.2058,found:415.2057.

Example 16

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (1-phenylethylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the procedure is as in example 7, using acetophenone instead of benzaldehyde, in 81% yield.

White solid, melting point 166-167 ℃.¹H NMR(400MHz,DMSO-d⁶)δ9.55(s,1H),7.82(d,J＝7.1Hz,2H),7.78(s,1H),7.37(t,J＝7.3Hz,2H),7.32(d,J＝7.0Hz,1H),4.26–4.21(m,1H),4.16–4.07(m,1H),3.24(s,3H),2.27(s,3H),2.15(d,J＝11.1Hz,1H),2.02–1.88(m,5H),1.82–1.76(m,1H),1.72–1.65(m,1H),1.58–1.47(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.98,156.02,151.51,143.84,139.09,138.39,128.04,127.92,125.47,115.71,61.42,59.59,28.49,28.40,27.65,26.25,23.57,23.41,12.97,8.69.HR-MS(ESI):Calcd.C₂₂H₂₈N₆O,[M+H]⁺m/z:393.2403,found:393.2402.

Example 17

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- (2-methoxybenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure used in example 7, except that 2-methoxybenzaldehyde was used instead of benzaldehyde, was found to be 81% in yield

White solid, melting point 220-222 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.72(s,1H),8.38(s,1H),7.88–7.82(m,1H),7.74(s,1H),7.34–7.26(m,1H),7.05(d,J＝8.2Hz,1H),6.96(t,J＝7.5Hz,1H),4.24–4.18(m,1H),4.12–4.03(m,1H),3.84(s,3H),3.23(s,3H),2.20–2.08(m,1H),2.07–1.86(m,5H),1.83–1.75(m,1H),1.71–1.63(m,1H),1.60–1.48(m,2H),0.82–0.71(m,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.95,156.78,155.25,151.33,138.48,134.82,129.65,124.67,123.80,120.45,115.50,111.58,61.68,59.64,55.52,28.64,28.50,27.63,26.23,23.97,23.84,8.68.HR-MS(ESI):Calcd.C₂₂H₂₈N₆O₂,[M+H]⁺m/z:409.2352,found:409.2352.

Example 18

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- (3-methoxybenzylidene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure used in example 7, except that 3-methoxybenzaldehyde was used instead of benzaldehyde, was found to be 81% in yield

White solid, melting point 181-182 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.75(s,1H),8.02(s,1H),7.76(s,1H),7.30(t,J＝7.9Hz,1H),7.21(s,1H),7.16(d,J＝7.7Hz,1H),6.94–6.86(m,1H),4.26–4.21(m,1H),4.16–4.08(m,1H),3.79(s,3H),3.23(s,3H),2.14–1.88(m,6H),1.83–1.76(m,1H),1.68(d,J＝7.1Hz,1H),1.59–1.47(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.93,159.50,155.19,151.40,139.27,138.53,137.14,129.64,119.00,115.61,114.51,110.07,61.47,59.44,54.98,28.70,28.63,27.64,26.24,23.97,23.87,8.71.HR-MS(ESI):Calcd.C₂₂H₂₈N₆O₂,[M+H]⁺m/z:409.2352,found:409.2351.

Example 19

(E) Preparation of-8-cyclopentyl-2- (2- (3, 4-dimethoxybenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure used in example 7, except that 3, 4-dimethoxybenzaldehyde was used in place of benzaldehyde, yield 81%

White solid, melting point 167-.¹H NMR(400MHz,DMSO-d⁶)δ10.59(s,1H),7.99(s,1H),7.75(s,1H),7.28(d,J＝1.7Hz,1H),7.11–7.02(m,1H),6.97(d,J＝8.4Hz,1H),4.24–4.15(m,2H),3.80(s,2H),3.78(s,2H),3.23(s,3H),2.08–1.88(m,6H),1.81–1.74(m,1H),1.69–1.62(m,1H),1.59–1.47(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.89,155.35,151.46,149.48,148.98,139.85,138.58,128.49,120.22,115.33,111.50,107.64,61.24,59.14,55.48,55.24,28.77,27.64,26.22,23.94,23.90,8.75.HR-MS(ESI):Calcd.C₂₃H₃₀N₆O3,

[M+H]⁺m/z:439.2457,found:439.2458.

Example 20

(E) Preparation of-8-cyclopentyl-2- (2- (3-ethoxy-2-hydroxybenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

using 2-hydroxy-3-ethoxybenzaldehyde instead of benzaldehyde, the same procedure as in example 7 was conducted, except that the yield was 81%

White solid, melting point 178-.¹H NMR(400MHz,DMSO-d⁶)δ11.98(s,1H),11.07(s,1H),8.20(s,1H),7.81(s,1H),6.93(t,J＝8.1Hz,2H),6.78(t,J＝7.8Hz,1H),4.54–4.36(m,1H),4.25–4.20(m,1H),4.03(q,J＝7.0Hz,2H),3.25(s,3H),2.05(d,J＝8.1Hz,1H),1.96–1.71(m,7H),1.70–1.60(m,3H),1.33(t,J＝7.0Hz,3H),0.77(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.83,154.72,151.84,147.45,146.93,141.24,138.68,121.34,119.46,118.48,115.91,114.86,28.82,28.32,27.75,26.46,22.66,22.48,14.82.HR-MS(ESI):Calcd.C₂₃H₃₀N₆O₃,[M+H]⁺m/z:439.2457,found:439.2458.

Example 21

(E) Preparation of-8-cyclopentyl-2- (2- (4- (diethylamino) -2-hydroxybenzylidene) hydrazino) -7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

using 2-hydroxy-4-diethylaminobenzaldehyde instead of benzaldehyde, the procedure is as in example 7, yield 81%

Yellow solid, melting point 223-225 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.80(s,1H),10.63(s,1H),8.06(s,1H),7.78(s,1H),7.03(d,J＝8.7Hz,1H),6.28–6.16(m,1H),6.07(d,J＝2.3Hz,1H),4.48–4.32(m,1H),4.24–4.12(m,1H),3.23(s,3H),2.06–1.98(m,1H),1.94(d,J＝16.1Hz,1H),1.87–1.70(m,5H),1.67–1.55(m,3H),1.09(t,J＝7.0Hz,6H),0.77(t,J＝7.5Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.79,158.98,155.01,151.88,148.92,143.04,138.86,130.57,

115.35,107.49,103.21,97.84,59.52,57.83,43.67,28.87,28.55,27.73,26.29,22.99,22.73,12.54,8.96.HR-MS(ESI):Calcd.C₂₅H₃₅N₇O₂,[M+H]⁺m/z:466.2930,found:466.2929.

Example 22

(E) Preparation of-2- ((2- (8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5, 6,7, 8-tetrahydropteridin-2-yl) hydrazono) methyl) benzonitrile:

the same procedure as in example 7 was carried out using 2-acetonitrile benzaldehyde instead of benzaldehyde, but the yield was 81%

White solid, melting point 155-.¹H NMR(400MHz,DMSO-d⁶)δ11.20(s,1H),8.41(s,1H),8.06(d,J＝8.0Hz,1H),7.83(d,J＝7.7Hz,1H),7.79(s,1H),7.71(t,J＝7.6Hz,1H),7.48(t,J＝7.6Hz,1H),4.27–4.22(m,1H),4.18–4.10(m,1H),3.24(s,3H),2.16–1.88(m,6H),1.83–1.76(m,1H),1.72–1.65(m,1H),1.57(d,J＝11.5Hz,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.99,154.68,151.38,138.60,138.38,134.33,133.15,133.08,128.56,124.58,117.32,116.26,109.24,61.43,59.51,28.56,28.47,27.68,26.31,23.86,23.75,8.66.HR-MS(ESI):Calcd.C₂₂H₂₅N₇O,[M+H]⁺m/z:404.2199,found:404.2198.

Example 23

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (naphthalen-1-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7, using 1-naphthaldehyde instead of benzaldehyde, gave a yield of 81%

White solid, melting point 157-.¹H NMR(400MHz,DMSO-d⁶)δ10.82(s,1H),9.02–8.93(m,1H),8.72(s,1H),8.01–7.95(m,1H),7.91(d,J＝8.1Hz,1H),7.82(t,J＝3.2Hz,2H),7.62–7.52(m,3H),4.38–4.31(m,1H),4.26–4.20(m,1H),3.26(s,3H),2.08–1.87(m,6H),1.82–1.75(m,1H),1.70–1.63(m,1H),1.62–1.54(m,2H),0.79(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.91,155.33,151.69,139.42,138.73,133.64,130.81,129.87,128.82,128.63,126.63,126.21,126.00,125.52,124.54,115.74,60.50,58.60,28.82,28.74,27.71,26.28,23.65,23.54,8.88.HR-MS(ESI):Calcd.C₂₅H₂₈N₆O,[M+H]⁺m/z:429.2403,found:429.2402.

Example 24

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((1E,2E) -3-phenylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the procedure is as in example 7, using cinnamaldehyde instead of benzaldehyde, in 81% yield.

Yellow solid, melting point 167-.¹H NMR(400MHz,DMSO-d⁶)δ10.65(s,1H),7.90(d,J＝9.1Hz,1H),7.74(s,1H),7.54(d,J＝7.4Hz,2H),7.37(t,J＝7.6Hz,2H),7.27(t,J＝7.3Hz,1H),6.99–6.90(m,1H),6.80(d,J＝16.1Hz,1H),4.33–4.25(m,1H),4.20–4.16(m,1H),3.23(s,3H),2.00–1.84(m,6H),1.78–1.71(m,1H),1.68–1.61(m,1H),1.60–1.50(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.87,155.03,151.49,141.97,138.65,136.50,134.27,128.73,127.96,126.56,126.48,115.72,60.31,58.34,28.79,28.73,27.71,26.31,23.65,23.43,8.88.HR-MS(ESI):Calcd.C₂₃H₂₈N₆O,[M+H]⁺m/z:405.2403,found:405.240.

Example 25

(E) Preparation of-2- (2- ((1H-indol-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used except that 1H-indole-2-carbaldehyde was used instead of benzaldehyde, and the yield was 60%.

White solid, melting point: 177 and 179 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.33(s,1H),10.37(s,1H),8.41(d,J＝7.8Hz,1H),8.25(s,1H),7.78(s,1H),7.62(d,J＝2.5Hz,1H),7.40(d,J＝8.1Hz,1H),7.17(t,J＝7.5Hz,1H),7.05(t,J＝7.3Hz,1H),4.59–4.46(m,1H),4.23–4.16(m,1H),3.24(s,3H),2.17–2.08(m,1H),2.03(s,1H),1.93–1.73(m,5H),1.70–1.56(m,3H),0.79(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.81,155.95,151.95,139.02,137.19,136.96,127.70,124.39,122.27,122.11,119.55,114.75,112.80,111.54,59.62,57.80,28.99,28.66,27.70,26.23,23.06,22.81.HR-MS(ESI):Calcd.C₂₃H₂₇N₇O,[M+H]⁺m/z:418.2355,found:418.2354.

Example 26

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (thien-2-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

thiophene-2-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7, giving a yield of 60%.

White solid, melting point 181-182 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.69(s,1H),8.25(s,1H),7.74(s,1H),7.50(d,J＝5.0Hz,1H),7.23(d,J＝2.8Hz,1H),7.11–6.99(m,1H),4.25–4.17(m,1H),4.14–4.03(m,1H),3.22(s,3H),2.13–1.89(m,6H),1.83–1.75(m,1H),1.70–1.64(m,1H),1.57(s,1H),0.75(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.92,155.03,151.34,140.77,138.49,134.80,127.56,127.49,126.54,115.54,61.59,59.62,28.67,28.53,27.63,26.25,24.08,23.93,8.69.HR-MS(ESI):Calcd.C₁₉H₂₄N₆OS,[M+H]⁺m/z:385.1810,found:385.1809.

Example 27

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((5-methylthiophen-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

5-methylthiophene-2-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7 in a yield of 60%.

White solid, melting point 177-178 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.59(s,1H),8.15(s,1H),7.73(s,1H),7.01(d,J＝3.5Hz,1H),6.79–6.68(m,1H),4.24–4.18(m,1H),4.13–4.04(m,1H),3.22(s,3H),2.43(s,3H),2.12–1.87(m,6H),1.83–1.75(m,1H),1.72–1.62(m,1H),1.60–1.48(m,2H),0.75(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.91,155.09,151.33,140.19,138.55,138.51,135.16,127.81,125.82,115.42,61.52,59.54,28.68,28.55,27.63,26.24,24.06,23.92,15.26,8.70.HR-MS(ESI):Calcd.C₂₀H₂₆N₆OS,[M+H]⁺m/z:399.1967,found:399.1968.

Example 28

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- (furan-2-ylmethylene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

furan-2-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7, giving a yield of 60%.

White solid, melting point 181-183 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.68(s,1H),7.99(s,1H),7.73(d,J＝9.1Hz,2H),6.66(d,J＝3.2Hz,1H),6.62–6.51(m,1H),4.24–4.14(m,2H),3.23(s,3H),2.00(d,J＝2.0Hz,2H),1.96–1.84(m,4H),1.80–1.73(m,1H),1.69–1.62(m,1H),1.52(d,J＝9.3Hz,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.90,155.05,151.36,150.85,143.44,138.53,130.00,115.70,111.87,109.13,60.98,58.96,54.88,28.72,28.68,27.67,26.30,23.84,23.75,8.77.HR-MS(ESI):Calcd.C₁₉H₂₄N₆O₂,[M+H]⁺m/z:369.2039,found:369.2038.

Example 29

(E) Preparation of-2- (2- ((1H-pyrrol-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

pyrrole-2-carbaldehyde was used instead of benzaldehyde in the same manner as in example 7 with a yield of 60%.

White solid, melting point 183-184 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.07(s,1H),10.27(s,1H),7.91(s,1H),7.65(s,1H),6.75(s,1H),6.25(s,1H),6.03(s,1H),4.26(d,J＝8.5Hz,1H),4.15–4.06(m,1H),3.22–3.12(m,3H),1.83(t,J＝16.7Hz,6H),1.71–1.64(m,1H),1.60–1.44(m,3H),0.71(t,J＝7.3Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.84,155.37,151.55,138.53,133.71,128.37,120.48,115.28,109.53,108.83,60.17,58.18,28.82,28.74,27.72,26.29,23.58,23.36,8.92.HR-MS(ESI):Calcd.C₁₉H₂₅N₇O,[M+H]⁺m/z:368.2199,found:368.2199.

Example 30

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (pyridin-2-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

pyridine-2-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7 in a yield of 70%.

White solid, melting point 188-.¹H NMR(400MHz,DMSO-d⁶)δ11.02(s,1H),8.53(d,J＝4.7Hz,1H),8.11(s,1H),7.89(d,J＝8.0Hz,1H),7.84–7.76(m,2H),7.36–7.21(m,1H),4.29–4.21(m,1H),4.14(s,1H),3.24(s,3H),2.10(d,J＝10.1Hz,1H),1.94(d,J＝21.8Hz,5H),1.84–1.76(m,1H),1.72–1.64(m,1H),1.62–1.50(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.97,154.88,154.64,151.40,149.17,139.87,138.46,136.29,122.83,118.52,116.04,61.40,59.44,28.64,28.55,27.68,26.30,23.89,23.81,8.69.HR-MS(ESI):Calcd.C₂₀H₂₅N₇O,[M+H]⁺m/z:380.2199,found:380.2198.

Example 31

(E) Preparation of-2- (2- ((6-chloropyridin-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used, using 6-chloropyridine-2-carbaldehyde instead of benzaldehyde, in a yield of 70%.

White solid, melting point 182 ℃ and 183 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.18(s,1H),8.02(s,1H),7.91–7.78(m,3H),7.40(d,J＝7.0Hz,1H),4.28–4.22(m,1H),4.20–4.10(m,1H),3.24(s,3H),2.11–1.91(m,6H),1.83–1.76(m,1H),1.72–1.64(m,1H),1.63–1.51(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.97,155.53,154.64,151.40,149.74,139.84,138.41,137.83,122.87,117.40,116.31,61.29,59.38,28.63,28.56,27.69,26.32,23.86,23.79,8.69.HR-MS(ESI):Calcd.C₂₀H₂₄ClN₇O,[M+H]⁺m/z:414.1809,found:414.1810.

Example 32

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((5-methylpyridin-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

5-methylpyridine-2-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7, and the yield was 70%.

White solid, melting point 191-193 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.94(s,1H),8.37(s,1H),8.08(s,1H),7.83–7.75(m,2H),7.62(d,J＝6.6Hz,1H),4.26–4.21(m,1H),4.18–4.09(m,1H),3.24(s,3H),2.30(s,3H),2.17–2.06(m,1H),2.05–1.89(m,5H),1.83–1.75(m,1H),1.72–1.64(m,1H),1.62–1.50(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.95,154.96,152.12,151.39,149.27,140.01,138.49,136.79,132.23,118.05,115.91,61.43,59.44,28.66,28.56,27.66,26.29,23.91,23.82,17.77,8.70.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O,[M+H]⁺m/z:394.2355,found:394.2356.

Example 33

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((6-methylpyridin-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used, using 6-methylpyridine-2-carbaldehyde instead of benzaldehyde, in a yield of 70%.

White solid, melting point 221-.¹H NMR(400MHz,DMSO-d⁶)δ11.00(s,1H),8.06(s,1H),7.78(s,1H),7.73–7.65(m,2H),7.19–7.11(m,1H),4.27–4.21(m,1H),4.20–4.09(m,1H),3.24(s,3H),2.46(s,3H),2.13–2.05(m,1H),2.05–1.87(m,5H),1.83–1.74(m,1H),1.72–1.64(m,1H),1.62–1.49(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.95,157.39,154.93,154.03,151.40,140.12,138.49,136.51,122.04,115.96,115.66,61.34,59.38,28.66,28.58,27.67,26.30,23.87,23.81,8.70.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O,[M+H]⁺m/z:394.2355,found:394.2356.

Example 34

(E) Preparation of-2- (2- ((4-chloropyridin-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used except that 4-chloropyridine-2-carbaldehyde was used in place of benzaldehyde in a yield of 70%.

White solid, melting point: 172-174 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.23(s,1H),8.50(d,J＝5.4Hz,1H),8.08(s,1H),7.90(d,J＝1.9Hz,1H),7.79(s,1H),7.45–7.40(m,1H),4.31–4.24(m,1H),4.12–3.98(m,1H),3.25(s,3H),2.26–2.17(m,1H),2.14–2.00(m,3H),1.98–1.89(m,2H),1.86–1.78(m,1H),1.76–1.67(m,1H),1.64–1.52(m,2H),0.79–0.72(m,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.00,156.57,154.60,151.31,150.75,142.97,138.25,137.83,122.55,117.95,116.23,62.05,60.10,28.55,28.34,27.65,26.30,24.12,23.87,8.57.HR-MS(ESI):Calcd.C₂₀H₂₄ClN₇O,[M+H]⁺m/z:414.1809,found:414.1810.

Example 35

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- ((6-methoxypyridin-2-yl) methylene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used except that 6-methoxypyridine-2-carbaldehyde was used in place of benzaldehyde in a yield of 70%.

White solid, melting point 183-184 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.01(s,1H),8.01(s,1H),7.78(s,1H),7.71(t,J＝7.8Hz,1H),7.50(t,J＝9.7Hz,1H),6.72(d,J＝8.1Hz,1H),4.26–4.21(m,1H),4.19–4.11(m,1H),3.87(s,3H),3.24(s,3H),2.15–2.05(m,1H),2.04–1.88(m,5H),1.84–1.76(m,1H),1.71–1.65(m,1H),1.60–1.50(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.26,162.95,154.88,152.41,151.39,139.42,139.13,138.46,116.00,111.64,109.28,61.39,59.41,52.93,28.66,28.58,27.67,26.29,23.92,23.84,8.70.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O₂,[M+H]⁺m/z:410.2304,found:410.2303.

Example 36

(E) Preparation of (E) -2- (2- ((5-chloropyridin-2-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one

5-chloropyridine-2-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7 in a yield of 70%.

White solid, melting point 185-187 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.11(s,1H),8.57(d,J＝2.3Hz,1H),8.10(s,1H),7.98–7.92(m,1H),7.87(d,J＝8.6Hz,1H),7.79(s,1H),4.27–4.22(m,1H),4.19–4.11(m,1H),3.24(s,3H),2.12–1.89(m,6H),1.83–1.76(m,1H),1.71–1.65(m,1H),1.62–1.51(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.96,154.72,153.28,151.40,147.65,138.48,138.44,136.30,129.69,119.59,116.21,61.34,59.40,28.64,28.56,27.68,26.31,23.88,23.81,8.70.HR-MS(ESI):Calcd.C₂₀H₂₄ClN₇O,[M+H]⁺m/z:414.1809,found:414.1808.

Example 37

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (pyridin-3-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

pyridine-3-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7 in a yield of 70%.

White solid, melting point 177-170 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.93(s,1H),8.78(d,J＝1.7Hz,1H),8.54–8.44(m,1H),8.07(s,1H),8.03–7.92(m,1H),7.77(s,1H),7.48–7.33(m,1H),4.28–4.20(m,1H),4.18–4.04(m,1H),3.24(s,3H),2.17–1.90(m,6H),1.84–1.76(m,1H),1.72–1.65(m,1H),1.61–1.48(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.95,155.01,151.36,148.99,147.50,138.46,136.18,132.28,131.53,123.75,115.87,61.59,59.58,28.65,28.53,27.65,26.27,23.97,23.86,8.68.HR-MS(ESI):Calcd.C₂₀H₂₅N₇O,[M+H]⁺m/z:380.2199,found:380.2200.

Example 38

(E) Preparation of-2- (2- ((2-chloropyridin-3-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used except that 2-chloropyridine-3-carbaldehyde was used instead of benzaldehyde, and the yield was 70%.

White solid, melting point 176-178 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.19(s,1H),8.40–8.33(m,2H),8.31–8.25(m,1H),7.78(s,1H),7.52–7.45(m,1H),4.27–4.22(m,1H),4.15–4.05(m,1H),3.24(s,3H),2.15–2.07(m,1H),2.02–1.87(m,5H),1.84–1.77(m,1H),1.72–1.65(m,1H),1.60–1.48(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ154.71,151.35,148.97,148.09,138.38,134.31,133.49,129.90,123.43,116.22,61.53,59.58,28.58,28.47,27.67,26.30,23.92,23.81,8.66.HR-MS(ESI):Calcd.C₂₀H₂₄ClN₇O,[M+H]⁺m/z:414.1809,found:414.1808.

Example 39

(E) Preparation of-2- (2- ((2-aminopyridin-3-yl) methylene) hydrazino) -8-cyclopentyl-7-ethyl-5-methyl-7, 8-dihydropteridin-6 (5H) -one:

the procedure of example 7 was repeated using 2-aminopyridine-3-carbaldehyde instead of benzaldehyde in a yield of 70%.

White solid, melting point 188-.¹H NMR(400MHz,DMSO-d⁶)δ10.77(s,1H),8.11(s,1H),7.96–7.90(m,1H),7.82(s,1H),7.58(s,2H),7.48–7.41(m,1H),6.64–6.54(m,1H),4.44–4.33(m,1H),4.22–4.15(m,1H),3.23(s,3H),2.03–1.97(m,1H),1.93(s,1H),1.82–1.70(m,5H),1.66–1.56(m,3H),0.77(t,J＝7.5Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.85,156.27,155.15,151.87,147.40,140.73,138.87,137.99,115.74,112.08,111.62,59.47,57.74,28.88,28.77,27.75,26.28,23.12,22.84,8.99.HR-MS(ESI):Calcd.C₂₀H₂₆N₈O,[M+H]⁺m/z:395.2308,found:395.2309.

Example 40

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (1- (pyridin-3-yl) ethylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the procedure is as in example 7, using 3-acetylpyridine instead of benzaldehyde, in a yield of 70%.

Pale yellow solid, melting point 181-183 ℃.¹H NMR(400MHz,DMSO-d⁶)δ9.74(s,1H),9.00(d,J＝1.8Hz,1H),8.56–8.44(m,1H),8.12(d,J＝8.1Hz,1H),7.79(s,1H),7.45–7.33(m,1H),4.28–4.20(m,1H),4.16–4.06(m,1H),3.25(s,3H),2.29(s,3H),2.16(d,J＝10.8Hz,1H),2.04–1.85(m,5H),1.83–1.75(m,1H),1.73–1.65(m,1H),1.60–1.46(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.00,155.83,151.48,148.58,146.90,141.56,138.34,134.51,132.50,123.12,115.91,61.49,59.68,28.48,28.38,27.66,26.27,23.59,23.43,12.81,8.67.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O,[M+H]⁺m/z:394.2355,found:394.2356.

EXAMPLE 41

(E) Preparation of-8-cyclopentyl-7-ethyl-2- (2- ((5-methoxypyridin-3-yl) methylene) hydrazino) -5-methyl-7, 8-dihydropteridin-6 (5H) -one:

5-methoxypyridine-3-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7, and the yield was 70%.

White solid, melting point 212 ℃ and 214 ℃.¹H NMR(400MHz,DMSO-d⁶)δ10.97(s,1H),8.35(s,1H),8.22(d,J＝2.6Hz,1H),8.07(s,1H),7.77(s,1H),7.56(s,1H),4.27–4.20(m,1H),4.18–4.07(m,1H),3.87(s,3H),3.24(s,3H),2.14–1.89(m,6H),1.84–1.76(m,1H),1.71–1.64(m,1H),1.61–1.48(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.94,155.48,154.99,151.39,140.20,138.46,137.62,136.03,132.29,115.86,115.08,61.52,59.49,55.38,28.69,28.63,27.65,26.27,24.01,23.91,8.69.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O₂,[M+H]⁺m/z:410.2304,found:410.230.

Example 42

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (pyridin-4-ylmethylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

pyridine-4-carbaldehyde was used in place of benzaldehyde in the same manner as in example 7 in a yield of 70%.

White solid, melting point 213-214 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.11(s,1H),8.59–8.50(m,2H),8.01(s,1H),7.79(s,1H),7.59–7.50(m,2H),4.28–4.21(m,1H),4.17–4.06(m,1H),3.24(s,3H),2.13(d,J＝2.8Hz,1H),2.05–1.90(m,5H),1.83–1.76(m,1H),1.72–1.65(m,1H),1.61–1.51(m,1H),0.76(t,J＝7.4Hz,2H).¹³C NMR(101MHz,DMSO-d⁶)δ162.98,154.75,151.37,149.92,142.87,138.39,136.38,119.93,116.21,61.54,59.59,28.98,28.62,28.50,27.68,26.30,23.97,23.85,8.67.HR-MS(ESI):Calcd.C₂₀H₂₅N₇O,[M+H]⁺m/z:380.2199,found:380.2200.

Example 43

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- (1- (pyridin-4-yl) ethylidene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used, using 4-acetylpyridine instead of benzaldehyde, in a yield of 70%.

White solid, melting point 224-226 ℃.¹H NMR(400MHz,DMSO-d⁶)δ9.91(s,1H),8.55(d,J＝5.9Hz,2H),7.81(s,1H),7.73(d,J＝6.0Hz,2H),4.30–4.22(m,1H),4.20–4.09(m,1H),3.25(s,3H),2.27(s,3H),2.20–2.11(m,1H),2.03–1.88(m,5H),1.83–1.76(m,1H),1.74–1.65(m,1H),1.56(d,J＝7.3Hz,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.02,155.50,151.54,149.55,146.04,141.08,138.13,119.64,116.21,61.35,59.62,28.43,28.36,27.69,26.31,23.56,23.39,12.37,8.66.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O,[M+H]⁺m/z:394.2355,found:394.2356.

Example 44

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((3-nitropyridin-4-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the procedure of example 7 was repeated except for using 3-nitropyridine-4-carbaldehyde in place of benzaldehyde in a yield of 70%.

White solid, melting point 212 ℃ and 214 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.59(s,1H),9.16(s,1H),8.77(d,J＝5.3Hz,1H),8.56(s,1H),8.04(d,J＝5.3Hz,1H),7.82(s,1H),4.32–4.22(m,1H),4.19–4.09(m,1H),3.25(s,3H),2.19–2.06(m,1H),1.94(t,J＝11.3Hz,5H),1.85–1.77(m,1H),1.74–1.65(m,1H),1.58(d,J＝7.9Hz,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ163.03,154.26,152.51,151.39,146.24,142.66,138.29,137.69,131.13,119.37,116.88,61.41,59.61,28.50,28.40,27.72,26.36,23.84,23.72,8.65.HR-MS(ESI):Calcd.C₂₀H₂₄N₈O₃,[M+H]⁺m/z:425.2049,found:425.2050.

Example 45

(E) Preparation of-8-cyclopentyl-7-ethyl-5-methyl-2- (2- ((4-methylpyridin-2-yl) methylene) hydrazino) -7, 8-dihydropteridin-6 (5H) -one:

the same procedure as in example 7 was used except that 4-methylpyridine-2-carbaldehyde was used instead of benzaldehyde, and the yield was 70%.

White solid, melting point 224-226 ℃.¹H NMR(400MHz,DMSO-d⁶)δ11.02(s,1H),8.38(d,J＝5.0Hz,1H),8.08(s,1H),7.76(d,J＝11.9Hz,2H),7.19–7.07(m,1H),4.31–4.21(m,1H),4.14–4.01(m,1H),3.24(s,3H),2.34(s,3H),2.20–2.12(m,1H),2.10–1.90(m,5H),1.86–1.77(m,1H),1.73–1.65(m,1H),1.63–1.52(m,2H),0.76(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO-d⁶)δ162.97,154.90,154.53,151.35,148.91,146.71,139.77,138.44,123.79,118.97,115.95,61.76,59.74,28.61,28.48,27.65,26.28,24.02,23.85,20.51,8.64.HR-MS(ESI):Calcd.C₂₁H₂₇N₇O,[M+H]⁺m/z:394.2355,found:394.2356.

Example 46

BRD4 protein inhibitory Activity assay

1. Experimental methods

An Anti-GST-Cryptate (Eu3+ Cryptate conjugated mouse monoclonal antibody Anti-glutamathione S-transferase) is used as an energy donor, a Streptavidin-d2(d2-conjugated Streptavidin) is used as an energy receptor, GST-BD1, Biotin-H4, Anti-GST-Cryptate, Streptavidin-d2 and Assay buffer with certain concentrations are respectively added into a 384-pore plate, the final reaction system is 20 mu L, the reaction is carried out for a certain time at room temperature, and finally the interaction strength of the two biomolecules is judged according to the ratio of a signal value at an emission wavelength of 665nm to a signal value at a wavelength of 620nm, so that the inhibitory activity of the small molecular compound on BRD4 protein is tested.

2. The experimental results are shown in the second table.

Example 47

MTT method tumor cell inhibitory Activity test

1. Experimental methods

The selected inventive compound and the positive control compound JQ1 are respectively prepared into solution with the concentration gradient of 50, 25, 12.5, 6.26, 3.125, 1.56, 0.78, 0.39 and 0.19 mu M by 1640 culture medium, and respectively matched with gastric cancer cells SGC, BGC, MGC and MKN45. The leukemia cells THP1 were co-cultured in 96-well plates for 5 days, with 1000 cells per well for gastric cancer cells and 5000 cells per well for leukemia cells. After the compound stimulates gastric cancer cells for 5 days, adding MTT solution into the cells, wherein each hole is 20 mul, continuously culturing for 4 hours in an incubator, then slightly sucking out the original culture medium, adding 150 mul DMSO for dissolving, and slightly shaking for 10 minutes at room temperature to ensure that crystals are completely dissolved; and taking out the 96-well plate, and measuring the light absorption value of each well by a microplate reader at 570 nm. Adding MTT solution into the cells after the compound stimulates leukemia cells for 5 days, wherein each well has 20 mu L, continuously culturing for 4 hours in an incubator, adding 100 mu L of triple solution into each well, reacting for 24 hours, measuring the light absorption value of each well by an enzyme-labeling instrument at 570nm, and calculating the survival rate of the cells and IC according to the light absorption value₅₀。

2. The experimental results are shown in Table III.

Claims

1. dihydropteridone compounds and pharmaceutically acceptable salts thereof are characterized in that, are the following compounds:

,

,

.

2 . The application of the dihydropteridone compound or its pharmaceutically acceptable salt according to claim 1 , wherein, it is used as an active ingredient to prepare a drug for gastric cancer or blood cancer. 3 .