CN111217815B - Compound containing pteridone skeleton and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the technical field of medicine, and relates to compounds of pteridone skeleton represented by general formula I, their preparation methods and pharmaceutical compositions in which the compounds represented by general formula I are active ingredients, wherein substituents R ₁ , R ₂ , R ₃ , R ₄ , A ¹ , A ² , A ³ have the meanings given in the specification. The present invention also relates to compounds of general formula I and optical isomers, pharmaceutically acceptable salts, solvates or prodrugs thereof and pharmaceutical compositions thereof in the manufacture of medicaments for the treatment and/or prevention of cancer and other proliferative diseases use.

Description

Compound containing pteridone skeleton and preparation method and application thereof

technical field

The invention belongs to the technical field of medicine, and relates to a compound containing a pteridone skeleton and a preparation method and application thereof, in particular to a compound containing a pteridone skeleton, its geometric isomers and pharmaceutically acceptable salts, solvates or pro- Medicines, methods for their preparation and pharmaceutical compositions containing said compounds. The present invention also relates to the use of such compounds for the preparation and/or prevention of medicaments for cancer and other proliferative diseases.

Background technique

Cancer is a malignant tumor, which is a deadly disease that seriously endangers human health. Worldwide, there are currently 14 million new cancer cases worldwide and 8.2 million cancer deaths. Among them, there were 1.8 million new lung cancer cases and 1.59 million deaths. Therefore, malignant tumor has become the second type of human killer after cardiovascular disease.

Polo-like kinases (PLKs) are a class of serine/threonine protein kinases that are highly conserved in structure and function, and play a crucial role in the mitotic phase of cells. The PLKs family contains five subtypes in the human body, namely PLK1, PLK2 (SNK), PLK3 (FNK or PRK), PLK4 (SAK) and PLK5, which play important regulatory roles in various stages of the cell cycle. The structure of PLKs contains two conserved regions: a highly conserved N-terminal and a C-terminal, where the N-terminal has a highly homologous serine/threonine kinase domain, and the C-terminal has the characteristics of regulating PLKs activity and subcellular dynamic localization Structural domain (polo-box domain, PBD). PLK1, PLK2, and PLK3 isoforms are expressed in all human tissues, but the distribution of PLK4 mRNA in adults is limited to tissues such as testis and thymus, and has unique physiological roles. The PLK1 gene was reported in 1994, located at 16p12, the mRNA is about 2.3kb, and the encoded protein is about 6.6×104. It is the most thoroughly studied PLK family member.

The human PLK1 kinase structure consists of two regions: the highly conserved N-terminal region, consisting of 252 amino acids; the C-terminal, consisting of two polo box structures, called the PBD region (polo box domain), each polo box is composed of one A special region composed of β6-α sequences. Although the overall crystal structure of PLK1 has not been reported, the crystal structures of both the N-terminal kinase domain and the C-terminal PBD domain have been successfully determined.

At present, many compounds targeting PLK1 have entered the clinical research stage, such as: BI6727, BI2536, Rigosertib (ON-01910), GSK461364, MLN0905, TAK-960, etc. BI6727 reported in the literature was researched and developed by Boehringer Ingelheim Company. It belongs to the pteridone class of compounds. It is a selective Polo-like kinases (PLKs) inhibitor obtained from compound virtual screening. The IC ₅₀ of PLK1 inhibition in vitro is 0.87nM. It can be seen that inhibiting the activity of PLK1 has anti-tumor effects both in vitro and in vivo. The study of small molecule inhibitors targeting PLK1 has opened up a new direction for the research and development of anti-tumor drugs, and has a good development prospect. Therefore, many scientific research institutions and commercial pharmaceutical companies have launched corresponding R&D programs.

Based on the references, the inventors designed and synthesized [1,2,4]triazolo[4,3-f]pteridine-4(5H)-one, 1-methyl-[1,2 ,4]Triazolo[4,3-f]pteridine-4(5H)-one, tetraazolo[4,3-f]pteridine-4(5H)-one three series of pteridones Derivatives were screened for anti-tumor activity of various tumor cell lines in vitro, and the results showed that they had anti-tumor activity.

SUMMARY OF THE INVENTION

The present invention relates to pteridone compounds represented by general formula (I) and geometric isomers, pharmaceutically acceptable salts, solvates or prodrugs thereof,

in,

A ¹ is N or CR ₁ , A ² is N or CR _1a , A ³ is N, C or CR _1b ,

wherein R ₁ , R _1a , R _1b , if present, are independently selected from hydrogen, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen or/and hydroxy or/and Amino-substituted (C ₁ -C ₆ )alkyl, halogen or/and hydroxy or/and amino-substituted (C ₁ -C ₆ )alkoxy;

R ₂ is (C ₁ -C ₆ )alkyl or (C ₃ -C ₈ )cycloalkyl;

R ₃ is (C ₁ -C ₆ ) alkyl, 3-12 membered saturated or partially unsaturated carbocyclic ring, or 4-7 membered heterocyclic ring containing N, O and/or S; or NH _n R ₄ R ₅ , NH _n COR ₄ R ₅ , NH _n CONH _n R ₄ R ₅ , SO ₂ (CH ₂ ) _n NR ₄ R ₅ , SO ₂ (CH ₂ ) _n CONR ₄ R ₅ ; they may be the same or different by 0-3 The R ₆ is substituted;

n is an integer from 0 to 2;

R ₄ and R ₅ are the same or different and are independently selected from hydrogen, (C ₁ -C ₁₀ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₁ -C ₄ ) alkoxy, (C 1 -C 4 ) alkoxy ₂ -C ₁₀ ) alkenyl, (C ₂ -C ₁₀ ) alkynyl;

or R ₄ and R ₅ together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group or a 4-10 membered heteroaryl group other than the one attached to R ₄ and R _{5 In} addition to nitrogen atoms, optionally containing _0-4 heteroatoms selected from N, O and/or S, and in addition to the nitrogen atoms connected to R4 and R5, the heterocyclic group optionally includes 0-2 carbons Carbon double bond or carbon-carbon triple bond, and the heterocyclic group or heteroaryl group is optionally substituted by 0-3 identical or different R ₆ ;

R ₆ is (C ₁ -C ₆ )alkyl, (C ₃ -C ₇ )cycloalkyl, (C ₁ -C ₆ )alkoxy, hydroxy, halogen, halo(C ₁ -C ₆ )alkyl , halogenated (C ₁ -C ₆ ) alkoxy, nitro.

The present invention preferably relates to the pteridone compounds represented by the general formula (I) and their geometric isomers, pharmaceutically acceptable salts, solvates or prodrugs thereof,

in,

A ¹ is N, A ² is N, A ³ is N, C or CR _1b ,

R _1b is independently selected from hydrogen, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen or/and hydroxy or/and amino substituted (C ₁ -C ₆ )alkane radical, halogen or/and hydroxy or/and amino substituted (C ₁ -C ₆ )alkoxy;

R ₂ is (C ₃ -C ₈ )cycloalkyl;

R ₃ is (C ₁ -C ₆ ) alkyl, 3-6 membered saturated or partially unsaturated carbocyclic ring, or 4-7 membered heterocyclic ring containing N, O and/or S; or NH _n R ₄ R ₅ , NH _n COR ₄ R ₅ , NH _n CONH _n R ₄ R ₅ , SO ₂ (CH ₂ ) _n NR ₄ R ₅ , SO ₂ (CH ₂ ) _n CONR ₄ R ₅ ; they may be the same or different by 0-3 The R ₆ is substituted;

n is an integer from 0 to 2;

R ₄ and R ₅ are the same or different, and are independently selected from hydrogen, (C ₁ -C ₆ ) alkyl, (C ₃ -C ₇ ) cycloalkyl, (C ₁ -C ₄ ) alkoxy, (C 1 -C 4 ) alkoxy ₂ - _C6 )alkenyl, (C2 _{- C6} )alkynyl;

Or R ₄ , R ₅ together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group or a 4-6 membered heteroaryl group, except that the heterocyclic group or heteroaryl group is attached to R ₄ and R ₅ . In addition to the nitrogen atom, it optionally contains 0-4 heteroatoms selected from N, O and/or S, and the heterocyclic group or heteroaryl group is optionally substituted by 0-3 identical or different R ₆ ;

R ₆ is (C ₁ -C ₆ )alkyl, (C ₃ -C ₇ )cycloalkyl, (C ₁ -C ₆ )alkoxy, hydroxy, halogen, halo(C ₁ -C ₆ )alkyl , halogenated (C ₁ -C ₆ ) alkoxy, nitro.

The present invention preferably relates to the pteridone compounds represented by the general formula (I) and their geometric isomers, pharmaceutically acceptable salts, solvates or prodrugs thereof,

in,

A ¹ is N, A ² is N, A ³ is N, C or CR _1b ,

R _1b is independently selected from hydrogen, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen or/and hydroxy or/and amino substituted (C ₁ -C ₆ )alkane radical, halogen or/and hydroxy or/and amino substituted (C ₁ -C ₆ )alkoxy;

R ₂ is (C ₃ -C ₆ )cycloalkyl;

_R3 is _R3 is

The present invention preferably relates to the pteridone compounds represented by the general formula (I) and their geometric isomers, pharmaceutically acceptable salts, solvates or their prodrugs, preferably the following compounds, but these compounds do not mean that the present invention any restrictions on:

5-Cyclopentyl-7-((4-(4-methylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine -4(5H)-one

5-Cyclopentyl-7-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine -4(5H)-one

5-Cyclopentyl-7-((4-(piperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine-4(5H )-ketone

5-Cyclopentyl-7-((4-(4-methylpiperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine -4(5H)-one

5-Cyclopentyl-7-((4-(pyrrolidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine-4(5H )-ketone

5-Cyclopentyl-7-((4-(3,5-dimethylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f ]Pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(4-thiomorpholinyl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine-4( 5H)-keto

5-Cyclopentyl-7-((4-(4-(2-hydroxyethyl)piperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3 -f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(4-hydroxypiperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine- 4(5H)-ketone

5-Cyclopentyl-7-((4-(cyclopentylamino)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine-4(5H)- ketone

5-Cyclopentyl-7-((2-methoxy-4-(4-ethylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4, 3-f]Pteridine-4(5H)-one

5-Cyclopentyl-7-((2-methoxy-4-(piperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f] pteridine-4(5H)-one

5-Cyclopentyl-7-((2-methoxy-4-(morpholin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f] pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(4-methylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3 -f]pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3 -f]pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(piperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteride Pyridin-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(4-methylpiperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3 -f]pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(pyrrolidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteran Pyridin-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(3,5-dimethylpiperazin-1-yl)phenyl)amino)-[1,2,4]triazolo[ 4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(4-thiomorpholinyl)phenyl)amino)-[1,2,4]triazolo[4,3-f] pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(4-(2-hydroxyethyl)piperazin-1-yl)phenyl)amino)-[1,2,4]triazole Ipo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(4-hydroxypiperidin-1-yl)phenyl)amino)-[1,2,4]triazolo[4,3- f]Pteridine-4(5H)-one

5-Cyclopentyl-1-methyl-7-((4-(cyclopentylamino)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine- 4(5H)-ketone

5-Cyclopentyl-7-((4-(4-methylpiperazin-1-yl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(piperidin-1-yl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(4-methylpiperidin-1-yl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(pyrrolidin-1-yl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(3,5-dimethylpiperazin-1-yl)phenyl)amino)tetrazolo[4,3-f]pteridine-4(5H) -ketone

5-Cyclopentyl-7-((4-(4-thiomorpholinyl)phenyl)amino)tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(4-(2-hydroxyethyl)piperazin-1-yl)phenyl)amino)tetrazolo[4,3-f]pteridine-4( 5H)-keto

5-Cyclopentyl-7-((4-(4-hydroxypiperidin-1-yl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(cyclopentylamino)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

5-Cyclopentyl-7-((4-(4-(4-methylpiperazin-1-yl)-piperidin-1-yl)phenyl)amino)-tetrazolo[4,3- f]Pteridine-4(5H)-one.

Moreover, according to some common methods in the art to which the present invention pertains, the pteridone skeleton-containing compound of the above formula I of the present invention can be formed into a pharmaceutically acceptable salt with an acid. Pharmaceutically acceptable addition salts include inorganic and organic acid addition salts, with addition salts with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid , benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc.

Furthermore, the present invention also includes prodrugs of the derivatives of the present invention. Prodrugs of the derivatives of the present invention are derivatives of general formula I, which themselves may have weak or even no activity, but after administration, are destroyed under physiological conditions (eg by metabolism, solvolysis or otherwise) by into the corresponding biologically active form.

In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to straight or branched chain alkyl; "cycloalkyl" refers to substituted or unsubstituted cycloalkyl; "Heteroaryl" refers to an unsubstituted or substituted phenyl group; "Heteroaryl" refers to a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, cyclic The system is aromatic, such as imidazolyl, pyridyl, pyrazolyl, (1,2,3)- and (1,2,4)-triazolyl, furyl, thienyl, pyrrolyl, thiazolyl, Benzothiazolyl, oxazolyl, isoxazolyl, naphthyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, etc.; "saturated or partially saturated heterocyclyl" means Monocyclic or polycyclic ring systems containing one or more heteroatoms selected from N, O, S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidine, imidazolidine base and thiazolinyl, etc.

The present invention can contain the compound containing pteridone skeleton of the above formula I, and its pharmaceutically acceptable salts and solvates as active ingredients, mixed with pharmaceutically acceptable carriers or excipients to prepare a composition, and prepared In order to form a clinically acceptable dosage form, the above-mentioned pharmaceutically acceptable excipients refer to any diluents, adjuvants and/or carriers that can be used in the pharmaceutical field. The derivatives of the present invention can be used in combination with other active ingredients as long as they do not produce other adverse effects, such as allergic reactions.

The clinical dosage of the compound containing the pteridone skeleton of the above formula I of the present invention for a patient can be determined according to: the therapeutic efficacy and bioavailability of the active ingredient in vivo, their metabolism and excretion rates, and the age, sex, and disease stage of the patient. Appropriate adjustments are made, but the daily dose for adults should generally be 10-500 mg, preferably 50-300 mg. Therefore, when the pharmaceutical composition of the present invention is formulated into a unit dosage form, considering the above-mentioned effective dose, each unit preparation should contain 10-500 mg, preferably 50-300 mg, of the compound containing the pteridone skeleton of the above formula I. These formulations may be administered in several divided doses (preferably one to six doses) at regular intervals, as directed by a physician or pharmacist.

The pharmaceutical composition of the present invention can be formulated into several dosage forms, which contain some commonly used excipients in the pharmaceutical field. Several of the above-mentioned dosage forms can be used as injections, tablets, capsules, aerosols, suppositories, films, drop pills, external liniments, ointments and other dosage forms.

The carriers used in the pharmaceutical compositions of the present invention are of the common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, cosolvents, diluents, stabilizers, suspending agents, flavoring agents, preservatives , solubilizers and substrates, etc. Pharmaceutical formulations may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically), and if certain drugs are unstable under gastric conditions, they may be formulated as enteric-coated tablets.

Through in vitro inhibition of human prostate cancer cell PC-3, human breast cancer cell MCF-7, human breast cancer cell MDA-MB-231, human colon cancer cell HCT116 and human lung adenocarcinoma cell A549 activity test, we found that the compound of the present invention has Significant antitumor activity, so the compounds of the present invention can be used for the preparation of medicines for the treatment and/or prevention of various cancers, such as breast, lung, liver, kidney, colon, rectum, stomach, prostate, bladder, uterus, pancreas, bone marrow, Cancer and leukemia of testis, ovary, lymph, soft tissue, head and neck, thyroid, esophagus, neuroblastoma, etc. In particular, it is used for the preparation of medicines for the treatment and/or prevention of lung cancer and liver cancer.

The active compounds of the present invention or their pharmaceutically acceptable salts and their solvates can be used alone as the only anti-tumor drugs, or can be used together with existing anti-tumor drugs (such as platinum drug cisplatin, camptothecin drug iritinib) Kang, vinblastine drugs Noviben, deoxycytidine drugs gemcitabine, etoposide, paclitaxel, etc.) are used in combination. Combination therapy is accomplished by administering the individual therapeutic components simultaneously, sequentially or separately.

The examples and preparations provided hereinafter further illustrate and illustrate the compounds of the present invention and methods for their preparation. It should be understood that the scope of the following examples and preparations do not limit the scope of the invention in any way.

The following synthetic schemes describe the preparation of the derivatives of general formula I of the present invention, all starting materials were prepared by methods described in these schemes, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available. All final compounds of the present invention are prepared by methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors applied in these routes are as defined below or as defined in the claims.

Route one:

Route two:

Route three:

Route four:

Detailed ways:

The following examples are intended to illustrate rather than limit the scope of the invention. The hydrogen nuclear magnetic resonance spectrum of the compound was measured with BrukerARX-400 or -600, and the mass spectrum was measured with Agilent 1100LC/M(S)D; all reagents used were of analytical or chemical purity.

Example 1 5-Cyclopentyl-7-((4-(4-morpholinyl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine-4 (5H)-keto

1.1 2-Chloro-4-cyclopentylamino-5-nitropyrimidine (2)

50 g (259 mmol) of 2,4-dichloro-5-nitropyrimidine, 43.5 g (518 mmol) of NaHCO ₃ , and 200 mL of DCM were added to a three-necked flask, and a solution of 26.4 g (311 mmol) of cyclopentylamine in DCM was added dropwise to the flask. In the above reaction solution, the reaction was carried out at room temperature for 10h. After the reaction was completed, the reaction solution was poured into 200 mL of water, the organic layer was separated, the aqueous layer was extracted twice with 100 mL of DCM, the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was evaporated to dryness, ether was added to the residue, and a pale yellow solid was precipitated. , filtered with suction, and dried to obtain 53.27 g of pale yellow solid, yield 85.0%, MS (ESI) m/z: 243.3 [M+H] ⁺ .

1.2 Synthesis of 2-chloro-4-cyclopentylamino-5-aminopyrimidine (3)

At room temperature, iron powder (57.8 g, 1033 mmol) was added to 95% ethanol (200 mL) with stirring, and the temperature was raised to 70° C. to activate the reaction for 30 min. 2-Chloro-4-cyclopentylamino-5-nitropyrimidine 2 (50 g, 207 mmol) was added to the reaction solution, and the temperature was raised to 80° C. to react for 2 h. After the reaction was completed, the mixture was filtered through a pad of diatomaceous earth while hot, and the solvent was evaporated, and purified by column chromatography to obtain 32.0 g of a white solid with a yield of 73%, MS (ESI) m/z: 213.2 [M+H] ⁺ .

1.3 Synthesis of 2-chloro-8-cyclopentyl-5,8-dihydropteridine-6,7-dione (4)

2-Chloro-4-cyclopentylamino-5-aminopyrimidine 3 (30 g, 124 mmol), K ₂ CO ₃ (34.2 g, 248 mmol) were dissolved in 150 mL of acetone at room temperature, and oxalyl chloride monoethyl ester (28.9 g, 136.4 mmol) was slowly added dropwise to the above reaction solution and reacted at room temperature for 2 h. Suction filtration, the filtrate was concentrated, the residue, 200 mL of EtOH, and TEA (15.1 g, 148.8 mmol) were added to a 500 mL pressure-resistant bottle, the temperature was raised to 120 ° C and stirred for 4 h. After the reaction was completed, the reaction solution was cooled to room temperature, suction filtered, and dried. 27.4 g of white solid was obtained, the yield was 83%, MS (ESI) m/z: 264.9 [MH] ⁻ .

1.4 Synthesis of 2,6-dichloro-8-cyclopentylpteridine-7(8H)-one (5)

2-Chloro-8-cyclopentyl-5,8-dihydropteridine-6,7-dione 4 (25 g, 94 mmol) was dissolved in 80 mL of SOCl ₂ , the temperature was raised to 80° C. and reacted for 3 h. After the reaction was completed, the The reaction solution was concentrated, the residue was added to a large amount of ice water, suction filtered, and dried to obtain 24.3 g of white solid, the yield was 91%, MS (ESI) m/z: 285.26 [M+H] ⁺ .

1.5 Synthesis of 2-chloro-8-cyclopentyl-6-hydrazinopteridine-7(8H)-one (6)

2,6-Dichloro-8-cyclopentylpteridine-7(8H)-one 5 (18 g, 63.4 mmol), 80% hydrazine hydrate (9.5 g, 190 mmol) were dissolved in 200 mL of EtOH, and the temperature was raised to 40 °C to react 2h. After the reaction was completed, the reaction solution was lowered to room temperature, filtered with suction, and dried to obtain 15.6 g of a white solid with a yield of 88%, MS(ESI) m/z: 279.0 [MH] ⁻ .

1.6 7-Chloro-5-cyclopentyl-[1,2,4]triazolo[4,3-f]pteridine-4(5H)-one (7)

2-Chloro-8-cyclopentyl-6-hydrazinopteridine-7(8H)-one 6 (6 g, 21.4 mmol) was added to 20 mL of triethyl orthoformate, and the temperature was raised to 80° C. and stirred for 2 h. After the reaction was completed, the reaction solution was cooled to room temperature, filtered with suction, and dried to obtain 5.3 g of a white solid with a yield of 86%, MS(ESI) m/z: 313.1 [M+Na] ⁺ .

1.7 Synthesis of 4-(4-nitrophenyl)morpholine (11)

P-fluoronitrobenzene, morpholine, K ₂ CO ₃ were added to DMF, and the temperature was raised to 80 °C and stirred for 2 h. After the reaction was completed, the reaction solution was cooled to room temperature, added to ice water, filtered with suction, and dried to obtain a yellow solid.

1.8 Synthesis of 4-morpholinoaniline (12)

4-(4-Nitrophenyl)morpholine and palladium on carbon (Pd/C) were added to 50 mL of ethanol, passed through with hydrogen, evacuated, and reacted at room temperature for 6 h. Filter with suction, wash the filter cake with ethanol (30 mL×3), discard the filter cake, and keep the filtrate. The filtrate solvent was removed.

1.9 5-cyclopentyl-7-((4-(4-morpholinyl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pteridine-4(5H )-ketone

7-Chloro-5-cyclopentyl-[1,2,4]triazolo[4,3-f]pteridine-4(5H)-one 7(1equiv), 12(1.1equiv), para Toluenesulfonic acid (1equiv) and isopropanol were added to a 50mL pressure bottle, and the temperature was raised to 100°C for reaction for 12h. After the reaction was completed, the reaction solution was cooled to room temperature, saturated sodium carbonate solution was added, extracted with 3×30 mL of dichloromethane, the organic phases were combined, dried, evaporated to remove the organic solvent, and purified by column chromatography to obtain pale yellow powder.

According to the method of Example 1, the compounds of Examples 1-13 were prepared by reacting intermediate 7 with various substituted anilines (see Table 1).

Table 1

Example 2 5-Cyclopentyl-1-methyl-7-((4-(4-morpholinyl)phenyl)amino)-[1,2,4]triazolo[4,3-f ]Pteridine-4(5H)-one

2.1 Synthesis of 7-chloro-5-cyclopentyl-1-methyl-[1,2,4]triazolo[4,3-f]pteridine-4(5H)-one (9)

According to the method of Example 1, intermediate 9 was prepared by reacting intermediate 6 with triethyl orthoacetate.

2.2 5-cyclopentyl-1-methyl-7-((4-(4-morpholinyl)phenyl)amino)-[1,2,4]triazolo[4,3-f]pyro Synthesis of Pyridin-4(5H)-one

7-Chloro-5-cyclopentyl-1-methyl-[1,2,4]triazolo[4,3-f]pteridine-4(5H)-one 7(1equiv), 12( 1.1 equiv), p-toluenesulfonic acid (1 equiv) and isopropanol were added to a 50 mL pressure bottle, and the temperature was raised to 100° C. for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, saturated sodium carbonate solution was added, extracted with 3×30 mL of dichloromethane, the organic phases were combined, dried, evaporated to remove the organic solvent, and purified by column chromatography to obtain pale yellow powder.

According to the method of Example 2, the compounds of Examples 14-23 were prepared by reacting intermediate 9 with various substituted anilines (see Table 2).

Table 2

Example 3 5-Cyclopentyl-7-((4-(4-morpholinyl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

2.1 Synthesis of 7-chloro-5-cyclopentyl-tetrazolo[4,3-f]pteridine-4(5H)-one (8)

According to the method of Example 1, intermediate 8 was prepared by reacting intermediate 5 with sodium azide.

2.2 Synthesis of 5-cyclopentyl-7-((4-(4-morpholinyl)phenyl)amino)-tetrazolo[4,3-f]pteridine-4(5H)-one

7-Chloro-5-cyclopentyl-tetrazolo[4,3-f]pteridine-4(5H)-one 8(1equiv), 12(1.1equiv), p-toluenesulfonic acid (1equiv) and Isopropanol was added to a 50 mL pressure-resistant bottle, and the temperature was raised to 100 °C for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, saturated sodium carbonate solution was added, extracted with 3×30 mL of dichloromethane, the organic phases were combined, dried, evaporated to remove the organic solvent, and purified by column chromatography to obtain pale yellow powder.

According to the method of Example 3, the compounds of Examples 24-34 were prepared by reacting intermediate 8 with various substituted anilines (see Table 3).

table 3

In vitro anti-tumor cell activity of the product of the present invention

In vitro inhibition of human prostate cancer cells PC-3, human breast cancer cells MCF-7, human breast cancer cells MDA-MB-231, and human colon cancer cells HCT116 was performed on the compounds of the pteridone skeleton of the above formula I according to the present invention. and human lung adenocarcinoma cell A549 activity assay. The reference substance BI6727 was prepared according to the method described in the literature (WO 2015180552).

(1) After the cells were recovered and passaged 2-3 times to stabilize, they were digested from the bottom of the culture flask with trypsin solution (0.25%). After the cell digestion solution was poured into the centrifuge tube, the culture solution was added to stop the digestion. Centrifuge the centrifuge tube at 800 r/min for 10 min, discard the supernatant, add 5 mL of culture medium, mix the cells by pipetting, pipette 10 μL of the cell suspension and add it to a cell counting plate for counting, and adjust the cell concentration to 10 ⁴ cells/well. In the 96-well plate, 100 μL of cell suspension was added to the rest of the 96-well plates, except that A1 was a blank well without cells. The 96-well plate was placed in an incubator for 24 h.

(2) Dissolve the test sample with 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into 2 mg/mL liquid, and then dilute the sample in a 24-well plate to 20, 4, 0.8, 0.16, 0.032 μg/mL.

The culture medium in the 96-well plate was discarded, and each concentration was added to 3 wells. The growth of the cells in the surrounding two rows and two columns was greatly affected by the environment, and was only used as a blank cell well. The 96-well plate was placed in an incubator for 72 h.

(3) 10 μL of MTT (0.5 mg/mL) was added to each well and placed in an incubator for 4 h, the MTT solution was discarded, and 100 μL of dimethyl sulfoxide was added. Shake on a magnetic shaker to fully dissolve the surviving cells and the MTT reaction product formazan, and put it into a microplate reader to measure the results. The drug IC ₅₀ value can be obtained by the Bliss method.

Table 4 shows the activity test results of some compounds in human prostate cancer cells PC-3, human breast cancer cells MCF-7, human breast cancer cells MDA-MB-231, human colon cancer cells HCT116 and human lung adenocarcinoma cells A549.

Table 4

It can be clearly seen from the above experimental results that the compound of general formula I to be protected by the present invention has good in vitro antitumor activity, which is comparable to or better than that of the positive control drug BI6727.

Claims (10)

1. A compound of general formula (I) and geometric isomers, pharmaceutically acceptable salts and solvates thereof,

wherein,

A¹is N, A²Is N, A³Is N, C or CR_1b，

Wherein R is_1bIndependently selected from hydrogen, halogen, (C), if present₁-C₆) Alkyl, (C)₁-C₆) Alkoxy, halogen and/or hydroxy and/or amino substituted (C)₁-C₆) Alkyl, halogen and/or hydroxy and/or amino substituted (C)₁-C₆) An alkoxy group;

R₂is (C)₁-C₆) Alkyl or (C)₃-C₈) A cycloalkyl group;

R₃is a 4-7 membered heterocyclic ring containing N, O and/or S; or NH_nR₄R₅、NH_nCONH_nR₄R₅、SO₂(CH₂)_nNR₄R₅、SO₂(CH₂)_nCONR₄R₅(ii) a They may be substituted by 0 to 3 identical or different R₆Substitution;

n is an integer of 0 to 2;

R₄and R₅The same or different, are respectively and independently selected from hydrogen and (C)₁-C₁₀) Alkyl, (C)₃-C₇) Cycloalkyl group, (C)₁-C₄) An alkoxy group;

or R₄And R₅Together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclyl or 4-10 membered heteroaryl group, other than R₄And R₅Optionally containing 0 to 4 heteroatoms selected from N, O and/or S in addition to the nitrogen atom to which it is attached, other than R₄And R₅Said heterocyclyl optionally including 0-2 carbon-carbon double or carbon-carbon triple bonds in addition to the attached nitrogen atom, said heterocyclyl or heteroaryl optionally being substituted with 0-3R, which may be the same or different₆Substitution;

R₆is (C)₁-C₆) Alkyl, (C)₃-C₇) Cycloalkyl group, (C)₁-C₆) Alkoxy, hydroxy, halogen, halo (C)₁-C₆) Alkyl, halo (C)₁-C₆) Alkoxy and nitro.

2. A compound of general formula (I) according to claim 1, and geometric isomers, pharmaceutically acceptable salts, solvates thereof,

wherein,

A¹is N, A²Is N, A³Is N, C or CR_1b，

R_1bIndependently selected from hydrogen, halogen, (C)₁-C₆) Alkyl, (C)₁-C₆) Alkoxy, halogen and/or hydroxy and/or amino substituted (C)₁-C₆) Alkyl, halogen and/or hydroxy and/or amino substituted (C)₁-C₆) An alkoxy group.

3. A compound of general formula (I) according to claim 1 or 2, and geometric isomers, pharmaceutically acceptable salts, solvates thereof,

wherein,

R₃is a 4-7 membered heterocyclic ring containing N, O and/or S; or NH_nR₄R₅、NH_nCONH_nR₄R₅、SO₂(CH₂)_nNR₄R₅、SO₂(CH₂)_nCONR₄R₅(ii) a They can be divided into 0-3Identical or different R₆Substitution;

n is an integer of 0 to 2;

R₄and R₅The same or different, are respectively and independently selected from hydrogen and (C)₁-C₆) Alkyl, (C)₃-C₇) Cycloalkyl group, (C)₁-C₄) An alkoxy group;

or R₄、R₅Together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclyl or 4-6 membered heteroaryl group, other than R₄And R₅Optionally containing 0 to 4 heteroatoms selected from N, O and/or S in addition to the nitrogen atom to which it is attached, said heterocyclyl or heteroaryl group optionally being substituted with 0 to 3R, which may be the same or different₆And (4) substitution.

4. A compound of general formula (I) according to any one of claims 1 to 3, and geometric isomers, pharmaceutically acceptable salts, solvates thereof,

wherein,

R₃is composed of

。

5. The following compounds and optical isomers, pharmaceutically acceptable salts and solvates thereof:

5-cyclopentyl-7- ((4- (4-methylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4-ethylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (piperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4-methylpiperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (pyrrolidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (3, 5-dimethylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-7- ((4- (4-thiomorpholinyl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4- (2-hydroxyethyl) piperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-7- ((4- (4-hydroxypiperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (cyclopentylamino) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((2-methoxy-4- (4-ethylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-7- ((2-methoxy-4- (piperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-7- ((2-methoxy-4- (morpholin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (4-methylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (4-ethylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (piperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H)-

Ketones

5-cyclopentyl-1-methyl-7- ((4- (4-methylpiperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (pyrrolidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (3, 5-dimethylpiperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Butterfly

Pyridine-4 (5)H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (4-thiomorpholinyl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (4- (2-hydroxyethyl) piperazin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]

Pteridine-4 (5)H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (4-hydroxypiperidin-1-yl) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine

-4(5H) -ketones

5-cyclopentyl-1-methyl-7- ((4- (cyclopentylamino) phenyl) amino) - [1,2,4]Triazolo [4,3-f]Pteridine-4 (5)H)-

Ketones

5-cyclopentyl-7- ((4- (4-methylpiperazin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4-ethylpiperazin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (piperidin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4-methylpiperidin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (pyrrolidin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (3, 5-dimethylpiperazin-1-yl) phenyl) amino) tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4-thiomorpholinyl) phenyl) amino) tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4- (2-hydroxyethyl) piperazin-1-yl) phenyl) amino) tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4-hydroxypiperidin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (cyclopentylamino) phenyl) amino) -tetrazolo [4,3-f]Pteridine-4 (5)H) -ketones

5-cyclopentyl-7- ((4- (4- (4-methylpiperazin-1-yl) -piperidin-1-yl) phenyl) amino) -tetrazolo [4,3-f]Pteridine

-4(5H) -a ketone.

6. A pharmaceutical composition comprising a compound of any one of claims 1-5 and pharmaceutically acceptable salts, solvates thereof as an active ingredient and a pharmaceutically acceptable excipient.

7. A process for the preparation of the compound of claim 1, wherein the intermediate is prepared as follows:

route one:

and a second route:

and a third route:

and a fourth route:

。

8. the use of a compound according to any one of claims 1 to 5, and pharmaceutically acceptable salts, solvates thereof, or pharmaceutical compositions according to claim 6, for the manufacture of a medicament for the treatment and/or prophylaxis of proliferative diseases.

9. Use of a compound according to any one of claims 1 to 5, and pharmaceutically acceptable salts, solvates thereof, or pharmaceutical compositions according to claim 6, for the manufacture of a medicament for the treatment and/or prophylaxis of cancer.

10. Use of a compound of any one of claims 1-5 and pharmaceutically acceptable salts, solvates thereof, or pharmaceutical composition of claim 6 for the preparation of a medicament for the treatment and/or prophylaxis of lung cancer, liver cancer, stomach cancer, colon cancer, breast cancer.