CN114181217B

CN114181217B - Pyrazolo [1,5-a ] [1,3,5] triazine derivative and salt thereof and application of derivative and salt thereof in preparation of anti-cancer drugs

Info

Publication number: CN114181217B
Application number: CN202210019208.2A
Authority: CN
Inventors: 李宝林; 席玮玮; 张娅玲; 王伟; 苗筱婉; 闫潇玉; 王子涵
Original assignee: Shaanxi Normal University
Current assignee: Shaanxi Normal University
Priority date: 2022-01-10
Filing date: 2022-01-10
Publication date: 2022-12-27
Anticipated expiration: 2042-01-10
Also published as: CN114181217A

Abstract

The invention discloses a pyridineAzolo [1,5-a][1,3,5]Triazine derivative and salt thereof and application of triazine derivative in preparation of anti-cancer drugs

Wherein R' is benzothiophen-3-yl or 1-methylindol-3-yl; r is-NHCH ₂ CH ₂ R 'diethylaminoethoxy, N, N, N' -trimethylethylenediamine, 4- (2-methoxyethyl) piperazinyl, diethylamino, 4-methylpiperazinyl, 4-acryloylpiperazinyl or

Wherein R' is diethylamino, morpholinyl, 4-methylpiperazinyl or pyrrolidinyl, R ¹ Is acrylamido, 4- (dimethylamino) but-2-enoylamino or N-methylpyrrolidin-2-ylimino; r ² Is one or two of fluorine, methoxy, dimethylamino, morpholinyl, diethylamino, thiomorpholinyl, 4-methylpiperazinyl and N, N, N' -trimethylethylenediamine; n is 1 or 2. Pyrazolo [1,5-a of the invention][1,3,5]The triazine derivative and the salt thereof have obvious inhibition effect on the proliferation of human cancer cells, and can be used for preparing anticancer drugs.

Description

Pyrazolo [1,5-a ] [1,3,5] triazine derivative and salt thereof and application of derivative and salt thereof in preparation of anti-cancer drugs

Technical Field

The invention belongs to the technical field of anti-cancer drugs, and particularly relates to novel pyrazolo [1,5-a ] [1,3,5] triazine derivatives and salts thereof, and application of the novel pyrazolo [1,5-a ] [1,3,5] triazine derivatives and salts thereof in preparation of anti-cancer drugs.

Background

According to recent reports around the world, lung cancer is the first ranked incidence of cancer, and about 80% to 85% of lung cancer patients are classified as non-small cell lung cancer (NSCLC). In the past, there have been surgical treatments, radiation treatments, chemotherapy treatments, etc. for NSCLC treatment. These treatments, while killing cancer cells, also cause irreversible damage to the patient's body. With the development of society and science, the cancer treatment means is continuously improved, and the toxic attack treatment on cells is developed into the targeted treatment aiming at cancer cells, so that the toxic and side effects of the medicament are greatly reduced, and the curative effect is obviously improved. After entering into the body, the targeted therapeutic drug can be specifically combined with a carcinogenic target to play a role, so that cancer cells are specifically killed, and normal tissue cells around the cancer cannot be affected. This method has been a hotspot in cancer therapy research. Among many cancer targets, epidermal Growth Factor Receptor (EGFR) Tyrosine Kinase (TK) is one of the cancer treatment targets with obvious effect and broad prospect.

Overexpression and abnormal activation of EGFR TK leads to excessive cell proliferation, leading to the development of cancer. Inhibiting the activity of EGFR TK, blocking the signal transduction pathway of cancer cell proliferation, and apoptosis. Thus, a range of EGFR TK inhibitors have been discovered and used in the treatment of cancer with good efficacy. The EGFR TK inhibitors can competitively inhibit the combination of Adenosine Triphosphate (ATP) and tyrosine kinase, inhibit the activity of tyrosine kinase, and inhibit the proliferation of cancer cells. Currently, small molecule tyrosine kinase inhibitors which are on the market and have better curative effect comprise Gefitinib (Gefitinib), lapatinib (Lapatinib), osertinib (Osimetinib) and Luo Xiti nib (Rociletinib), and although the drugs play a good role in the process of treating cancer, the drugs can generate drug resistance with different degrees after being used for a long time, so that the drug effect is reduced. Therefore, there is still a great need to find new low-toxicity, highly effective cancer therapeutic drugs.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a novel pyrazolo [1,5-a ] [1,3,5] triazine derivative with anticancer activity and salt thereof, and application of the compounds in preparing anticancer drugs.

The pyrazolo [1,5-a ] [1,3,5] triazine compound provided for solving the technical problems has the structural formula shown as follows:

in the formula, R' is any one of benzothiophen-3-yl and 1-methylindol-3-yl; r is-NHCH ₂ CH ₂ R ', diethylaminoethoxy, N, N' -trimethylethylenediamine, 4- (2-methoxyethyl) piperazinyl, diethylamino, 4-methylpiperazinyl, 4-acryloylpiperazinyl,

Wherein R' is any one of diethylamino, morpholinyl, 4-methylpiperazinyl and pyrrolidinyl, and R ¹ Is any one of acrylamido, 4- (dimethylamino) butyl-2-enamido and N-methylpyrrolidin-2-ylimino; r is ² Is a substituent attached to any position of the benzene ring, R ² Is selected from one or two of fluorine, methoxy, dimethylamino, morpholinyl, diethylamino, thiomorpholinyl, 4-methylpiperazinyl and N, N, N' -trimethylethylenediamine, and N is 1 or 2.

The salt of the pyrazolo [1,5-a ] [1,3,5] triazine derivative is a salt of the pyrazolo [1,5-a ] [1,3,5] triazine derivative with hydrochloric acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, maleic acid, fumaric acid, formic acid, acetic acid and benzoic acid. The salt is preferably a p-methylbenzenesulfonate salt of a pyrazolo [1,5-a ] [1,3,5] triazine derivative formed by reacting a pyrazolo [1,5-a ] [1,3,5] triazine derivative with p-methylbenzenesulfonic acid.

The pyrazolo [1,5-a ] [1,3,5] triazine derivative and the salt thereof are more preferably any of compounds 1 to 36, compound 1:2- (2-diethylaminoethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 2:2- (2-morpholinylethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 3:2- (2- (4-methylpiperazino) ethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 4:2- (4- (2-methoxyethyl) piperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 5:2- (4-methylpiperazino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 6: 2-piperazinyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 7:2- (4-acryloylpiperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 8:2- (4-methylpiperazino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine p-methylbenzenesulfonate; compound 9:2- (2-pyrrolidinylethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 10:2- (2-diethylaminoethoxy) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 11:2- (2-diethylaminoethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 12:2- (2-morpholinylethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 13:2- (4- (2-methoxyethyl) piperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 14:2- (4-methylpiperazino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 15: 2-piperazinyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 16:2- (4-acryloylpiperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 17:2- (2-pyrrolidinoethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 18: 2-diethylamino-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 19:2- (N, N' -trimethylethylenediamine) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 20:2- (3-acrylamido-4- (4-methylpiperazino) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 21:2- (3-acrylamido-4-diethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 22:2- (3-acrylamido-4-morpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 23:2- (3-acrylamido-4-thiomorpholino phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 24:2- (3-acrylamido-4- (N, N' -trimethylethylenediamine) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 25:2- (3-acrylamido-4-morpholinylphenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 26:2- (3-acrylamido-4- (4-methylpiperazino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 27:2- (3-acrylamido-4- (N, N' -trimethylethylenediamine) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 28:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 29:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 30:2- (3-acrylamido-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 31:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoroanilino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 32:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoroanilino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 33:2- (3-acrylamido-4- (N, N' -trimethylethylenediamine) -6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 34:2- (3-acrylamido-4-dimethylamino-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 35:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoro-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine; compound 36:2- (3- (N-methylpyrrolidin-2-yl) imino-4-fluoro-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine. The corresponding chemical structural formulas of the compounds 1 to 36 are as follows:

pyrazolo [1,5-a of the invention][1,3,5]In the synthesis of triazine derivatives 1-19, firstly, ethyl isothiocyanatocarboxylate and 2H-3-aminopyrazole are used as initial raw materials, and 1H-3 is generated through two-step reactionH-pyrazolo [1,5-a][1,3,5]Triazine-4-one parent nucleus, then with CH ₃ I reacts to generate 2-methylthio-3H-pyrazolo [1,5-a][1,3,5]Triazin-4-one with POCl in the presence of N, N-diethylaniline ₃ The reaction produces 2-methylthio-4-chloropyrazolo [1,5-a][1,3,5]A triazine having the formula:

when R' is benzothien-3-yl, 2-methylthio-4-chloropyrazolo [1,5-a][1,3,5]Triazine and benzothiophene-3-boronic acid in tris (dibenzylideneacetone) dipalladium (Pd) ₂ (dba) ₃ ) And 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (X-Phos) in the presence of K ₂ CO ₃ Carrying out Suzuki coupling reaction in a mixed system formed by aqueous solution and toluene to generate 2-methylthio-4- (benzothiophen-3-yl) pyrazolo [1,5-a][1,3,5]A triazine having the formula:

when R' is 1-methylindol-3-yl, 2-methylthio-4-chloropyrazolo [1,5-a][1,3,5]Triazines and 1-methylindoles with AlCl ₃ The catalyst is added into glycol dimethyl ether to react at 64 ℃ to generate 2-methylmercapto-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine having the formula:

further by oxidation of 2-methylsulfanyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine or 2-methylsulfanyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine with m-chloroperoxybenzoic acid (m-CPBA) in methylene chloride at room temperature to form the intermediate 2-methanesulfonyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine or 2-methanesulfonyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine, respectively, of the formula:

wherein R' is benzothiophen-3-yl or 1-methylindol-3-yl.

When R is-NHCH ₂ CH ₂ R' is derived from the intermediate 2-methanesulfonyl-4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine or 2-methanesulfonyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine reacts with 2-diethylaminoethylamine, 2-morpholinylethylamine, 2- (4-methylpiperazinyl) ethylamine and 2-pyrrolidinylethylamine in dioxane respectively to generate the triazine, and the reaction formula is as follows:

r' in the reaction formula is any one of diethylamino, morpholinyl, 4-methylpiperazinyl and pyrrolidinyl.

When R is diethylaminoethoxy, the intermediate 2-methanesulfonyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine is prepared by reacting 2-diethylaminoethanol in dioxane in the presence of metallic sodium, and the reaction formula is as follows:

when R is N, N, N '-trimethylethylenediamine, 4- (2-methoxyethyl) piperazinyl, diethylamino or 4-methylpiperazinyl, the intermediate obtained is obtained by reacting 2-methanesulfonyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine or 2-methanesulfonyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine with N, N, N' -trimethylethylenediamine, 4- (2-methoxyethyl) piperazine, diethylamine or N-methylpiperazine, respectively, in dioxane.

When R is 4-acryloylpiperazinyl, the compound is prepared by acylation reaction of 2-piperazinyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine or 2-piperazinyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine obtained by the above reaction with acryloyl chloride.

Pyrazolo [1,5-a of the invention][1,3,5]In the synthesis of triazine derivatives 20-36, when R is

In the synthesis process, a compound I (1-methylindole-3-formic acid or benzothiophene-3-formic acid) and oxalyl chloride react to generate a compound II; reacting the compound II with potassium thiocyanate to generate a compound III; the compound III reacts with 5-amino-1H-pyrazole to generate thiourea compound IV, and the reaction formula is as follows:

under the existence of mercuric chloride and triethylamine (or sodium hydride), the derivative of thiourea compound IV and 3-nitroaniline is obtained

Reacting in N, N-Dimethylformamide (DMF) under heating condition to generate a compound V; reducing the nitro group of the compound V under the action of hydrazine hydrate or iron powder to obtain a compound VI; the compound VI and acyl chloride are subjected to acylation reaction at room temperature to obtain the final target compound pyrazolo [1,5-a][1,3,5]Triazine derivative VII, having the formula:

in the acylation of the compounds VI with acid chlorides, if N-methylpyrrolidone is added, the acylation product is formed and at the same time R in the formula VII is formed ¹ Compound 36 which is N-methylpyrrolidin-2-ylimino.

The pyrazolo [1,5-a ] [1,3,5] triazine derivative and the salt thereof have obvious inhibition effect on the proliferation of human cancer cells, and can be used for preparing anticancer drugs. It is prepared into tablet, granule, capsule, injection, etc. with pharmaceutically acceptable carrier according to conventional preparation process. Wherein the cancer is any one of squamous carcinoma of skin, colon cancer and lung cancer.

The invention has the following beneficial effects:

the pyrazolo [1,5-a ] [1,3,5] triazine derivative and the salt thereof have simple synthesis methods, have obvious inhibition effects on the proliferation of human skin squamous carcinoma cells A431, human colon cancer cells SW480, human non-small cell lung cancer cells A549, human lung cancer cells NCI-H1975 and the like, can be used for preparing anticancer drugs, and can be used independently or combined with other drugs.

Detailed Description

The present invention will be described in further detail with reference to examples, but the scope of the present invention is not limited to these examples.

The reagents used in the following examples are all analytical grade. The nuclear magnetic resonance data used for compound structure identification is measured by a Bruker Avance 600 superconducting nuclear magnetic resonance instrument and a Bruker Avance 400 superconducting nuclear magnetic resonance instrument, and TMS is used as an internal standard; measuring infrared spectrum data by adopting a Bruker Tensor 2733 infrared spectrometer; the melting point was measured using an X-6 micro melting point apparatus (Beijing Takker instruments Co., ltd.) (temperature was not corrected); mass spectral data were measured with a Bruker Esquire 3000plus mass spectrometer.

Synthesis of 2-methylthio-4-chloropyrazolo [1,5-a ] used in the reaction][1,3,5]The triazine was synthesized according to the following method:

under ice bath conditions, 5.9g (0.07 mol) of 2H-3-aminopyrazole and 50mL of dichloromethane are added into a round-bottom flask, 30mL of dichloromethane solution containing 9.3g (0.07 mol) of isothiocyanatoethyl formate is slowly added, 80mL of dichloromethane is added for dilution, stirring is carried out for 2H, and the solvent is evaporated under reduced pressure to obtain a pale yellow solid. The resulting pale yellow solid was dissolved in 147mL of 2mol/L NaOH aqueous solution, stirred for 2 hours, and 211mL of 2mol/L H was added ₂ SO ₄ Dissolving in waterLiquid, white precipitate is generated immediately, after the mixture is fully stirred, the mixture is filtered, and a filter cake is dried to obtain a white solid product 1H-3H-pyrazolo [1,5-a][1,3,5]Triazine-4-one 10.6g, yield 90.6%, structural characterization data: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):13.47(s,1H),12.72(s,1H),7.88(br s,1H),5.90(br s,1H)。

1.0g (5.9 mmol) of 1H-3H-pyrazolo [1,5-a is weighed][1,3,5]Triazine-4-ketone in flask, adding 25mL ethanol to dissolve the solid, then adding 6.4mL 2mol/L NaOH aqueous solution, stirring slowly adding 0.9g (6.2 mmol) CH ₃ I, filtering after 2H, dissolving the filter cake with 39.6mL of water, and adding 3.9mL of 2mol/L H ₂ SO ₄ The water solution generates white precipitate immediately, the mixture is filtered after being fully stirred, and a filter cake is dried to obtain white solid 2-methylthio-3H-pyrazolo [1,5-a][1,3,5]Triazin-4-one 0.8g, 74.5% yield, structural characterization data: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):12.88(s,1H),7.98(d,J＝1.8Hz,1H),6.36(d,J＝1.8Hz,1H),2.54(s,3H)。

547mg (3 mmol) of 2-methylthio-3H-pyrazolo [1,5-a are weighed][1,3,5]Triazin-4-one in a flask, 5mL POCl was added ₃ And 175. Mu. L N, N-diethylaniline, were heated to reflux at 110 ℃ for 4h. Cooling, distilling under reduced pressure to remove solvent, adding 25mL toluene to dissolve residue, washing organic phase with saturated sodium chloride aqueous solution for 2 times, and evaporating solvent with rotary evaporator to obtain 2-methylthio-4-chloropyrazolo [1,5-a ]][1,3,5]Triazine 521mg, with a yield of 86.6%, and structural characterization data as follows: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):8.15(d,J＝2.1Hz,1H),6.50(d,J＝2.1Hz,1H),2.60(s,3H)。

the synthesis of 2-methanesulfonyl-4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine used in the following examples 1 to 10 is as follows:

weighing 2-methylthio-4-chloropyrazolo [1,5-a][1,3,5]Triazine 441mg (2.2 mmol), benzothiophene-3-boronic acid 263mg (1.5 mmol), tris (dibenzylideneacetone) dipalladium 34mg (0.037 mmol), 2-dicyclohexylphosphine-2 ',4',6' -triiso-lPropyl biphenyl 70mg (0.15 mmol) in a flask under nitrogen atmosphere 5.8mL 2mol/L of K ₂ CO ₃ The aqueous solution and 15mL of toluene were reacted at 60 ℃ for 12h. After the completion of the reaction, the solvent was distilled off by a rotary evaporator, and the residue was subjected to silica gel column chromatography (ethyl acetate: petroleum ether =1, 45, V/V) to give 2-methylthio-4- (benzothien-3-yl) pyrazolo [1,5-a as a pale yellow solid][1,3,5]Triazine 342mg, with a yield of 78.2%, and structural characterization data as follows: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.96(s,1H),9.09(d,J＝8.3Hz,1H),8.18(d,J＝2.1Hz,1H),7.95(d,J＝8.1Hz,1H),7.57(t,J＝7.3Hz,1H),7.48(t,J＝7.3Hz,1H),6.52(d,J＝2.1Hz,1H),2.70(s,3H)。

weighing 2-methylthio-4- (benzothiophen-3-yl) pyrazolo [1,5-a][1,3,5]298mg (1 mmol) of triazine are placed in a round-bottomed flask, 15mL of dichloromethane are added, 344mg (2 mmol) of m-chloroperoxybenzoic acid are slowly added under ice-bath conditions, and then stirring is carried out at ambient temperature for 4h. TLC monitoring reaction, after the reaction is completed, adding 15mL dichloromethane for dilution, washing with 2mol/L NaOH aqueous solution for 2 times, washing with saturated sodium chloride solution for 2 times, concentrating, and separating the residue by silica gel column chromatography (dichloromethane) to obtain light yellow solid 2-methylsulfonyl-4- (benzothiophen-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 238mg, yield 72.0%, structural characterization data: ¹ H NMR(400MHz,CDCl ₃ )δ(ppm):10.27(s,1H),9.29(d,J＝8.0Hz,1H),8.48(d,J＝2.3Hz,1H),7.97(d,J＝8.0Hz,1H),7.65(m,1H),7.53(m,1H),7.01(d,J＝2.3Hz,1H),3.48(s,3H)。

the synthesis of 2-methanesulfonyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine used in examples 11-19 below is as follows:

weighing 2-methylthio-4-chloropyrazolo [1,5-a][1,3,5]Triazine 482mg (2.4 mmol), alC1 ₃ 160mg (1.2 mmol) and 262mg (2.0 mmol) of 1-methylindole are placed in a round-bottomed flask. 10mL of ethylene glycol dimethyl ether (DME) was added under nitrogen atmosphere and reacted at 64 ℃ for 12 hours. After the reaction is completed, theThe reaction mixture is cooled to room temperature, filtered, the filter cake is washed 3 times with 20mL of methanol, the organic phases are combined and concentrated to dryness. The residue was subjected to silica gel column chromatography (dichloromethane) to give 2-methylsulfanyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a as a pale yellow solid][1,3,5]Triazine 532mg, yield 90.1%, structural characterization data: ¹ H NMR(400MHz,CDCl ₃ )δ(ppm):9.16(s,1H),8.71(d,J＝6.7Hz,1H),8.00(br s,1H),7.29(m,3H),6.31(br s,1H),3.83(s,3H),2.62(s,3H)。

weighing 2-methylthio-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]431mg (1.5 mmol) of triazine are dissolved in a round-bottom flask with 15mL of dichloromethane, 511mg (3 mmol) of m-chloroperoxybenzoic acid are added slowly under ice-bath conditions, and stirring is carried out at ambient temperature for 4h. Diluting with 15mL of dichloromethane, washing with 2mol/L NaOH aqueous solution for 2 times, washing with saturated sodium chloride solution for 2 times, concentrating, and separating the residue by silica gel column chromatography (dichloromethane) to obtain light yellow solid 2-methanesulfonyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 377mg, yield 79.0%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.46(s,1H),8.92(d,J＝8.1Hz,1H),8.38(br s,1H),7.53–7.39(m,3H),6.89(br s,1H),4.04(s,3H),3.51(s,3H)。

the synthesis of 1-methylindole-3-formyl-pyrazol-5-ylthiourea and benzothiophen-3-formyl-pyrazol-5-ylthiourea used in examples 20 to 36 below was carried out as follows:

weighing 876mg (5.0 mmol) of 1-methylindole-3-carboxylic acid into a round-bottom flask, adding 15mL of dichloromethane and catalytic amount of DMF (2 drops), slowly adding 0.6mL of oxalyl chloride under ice-bath condition in nitrogen atmosphere, stirring at room temperature for 4h after dropwise addition, evaporating to remove the solvent, adding 25mL of dried acetone and 729mg (7.5 mmol) of potassium thiocyanate into the residue, stirring at room temperature for 2h, evaporating to remove the solvent, adding 50mL of dichloromethane, washing with saturated aqueous sodium bicarbonate solution for 2 times, washing with saturated aqueous sodium chloride solution for 2 times, drying the organic phase with anhydrous sodium sulfate, and dryingEvaporated to dryness, dissolved in dichloromethane, added with 374mg (4.5 mmol) of 5-amino-1H-pyrazole, and stirred for 4H. The solid was filtered off and dried to yield 960mg of 1-methylindole-3-formyl-pyrazol-5-ylthiourea in 72.0% yield with structural characterization data: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):13.53(s,1H),12.75(s,1H),11.10(s,1H),8.76(s,1H),8.19(d,J＝7.8Hz,1H),7.73(s,1H),7.60(d,J＝8.1Hz,1H),7.35–7.31(m,1H),7.28(t,J＝7.1Hz,1H),7.10(s,1H),3.89(s,3H)。

the 1-methylindole-3-carboxylic acid in the synthesis method of the 1-methylindole-3-formyl-pyrazole-5-thiosemicarbazide is replaced by equivalent mass of benzothiophene-3-carboxylic acid, and other steps are the same as the steps to obtain 1.0g of benzothiophene-3-formyl-pyrazole-5-thiosemicarbazide, wherein the yield is 73.5 percent, and the structural characterization data are as follows: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):13.17(s,1H),12.81(s,1H),11.69(s,1H),8.97(s,1H),8.43(d,J＝8.0Hz,1H),8.12(d,J＝8.0Hz,1H),7.76(s,1H),7.54(t,J＝7.6Hz,1H),7.49(t,J＝7.4Hz,1H),7.11(s,1H)。

example 1

Weighing 2-methylsulfonyl-4- (benzothiophen-3-yl) pyrazolo [1,5-a][1,3,5]100mg (0.3 mmol) of triazine and 251mg (1.5 mmol) of 2-diethylaminoethylamine were placed in a round-bottomed flask, 5mL of dioxane were added, and heating and refluxing were carried out at 110 ℃ for 4 hours. The solvent was evaporated under reduced pressure and separated by silica gel column chromatography (methanol: dichloromethane =1, 40,v/V) to give compound 1 as a pale yellow solid: 2- (2-diethylaminoethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 90mg, yield 81.8%, structural characterization data: m.p.79.2-80.0 ℃; HRMS (C) ₁₉ H ₂₂ N ₆ S)m/z[M+H] ⁺ 367.1691 (calculated 367.1699); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.66(br s,1H),8.89(br s,1H),7.96(s,1H),7.90(s,1H),7.49(s,1H),7.42(s,1H),6.11(s,1H),6.05(s,1H),3.57(br s,2H),2.74(br s,2H),2.61(br s,4H),1.05(br s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.37,152.36,150.00,146.88,139.96,139.55,137.38,125.56,125.25,125.14,124.78,122.54,92.00,51.37,46.74,39.10,11.70；IR ν _max (KBr)cm ^-1 :3274,3102,2964,2921,1625,1522,1452,1270,908,761,726。

example 2

The same procedure as in example 1 was repeated except for replacing 2-diethylaminoethylamine in example 1 with an equivalent amount of 2-morpholinoethylamine to give a pale yellow solid compound 2:2- (2-Morpholinylethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 92mg, yield 80.7%, structural characterization data: m.p.137.0-138.4 deg.C; HRMS (C) ₁₉ H ₂₀ N ₆ OS)m/z[M+H] ⁺ 381.1492 (calculated 381.1492); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.71(s,1H),8.93(s,1H),8.01(d,J＝1.4Hz,1H),7.95(d,J＝8.0Hz,1H),7.58–7.51(m,1H),7.51–7.43(m,1H),6.15(s,1H),5.90(s,1H),3.76(d,J＝4.0Hz,4H),3.65(br s,2H),2.70(t,J＝5.7Hz,2H),2.55(br s,4H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.27,152.25,150.15,147.04,140.00,139.60,137.33,125.42,125.27,125.21,124.67,122.64,92.07,66.93,57.02,53.42,37.89；IR ν _max (KBr)cm ^-1 :3261,3033,2886,1618,1455,1260,911,732。

example 3

The same procedure as in example 1 was repeated except for replacing 2-diethylaminoethylamine in example 1 with an equivalent amount of 2- (4-methylpiperazinyl) ethylamine to give a pale yellow solid compound 3:2- (2- (4-methylpiperazino) ethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 82mg, yield69.5%, structural characterization data: m.p.140.7-141.3 ℃; HRMS (C) ₂₀ H ₂₂ N ₇ S)m/z[M+H] ⁺ 394.1809 (calculated 394.1808); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.70(s,1H),8.90(s,1H),7.98(s,1H),7.92(d,J＝7.6Hz,1H),7.49(t,J＝7.0Hz,1H),7.43(t,J＝6.8Hz,1H),6.13(s,1H),5.98(s,1H),3.63(br s,2H),2.91–2.53(m,10H),2.42(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.22,152.19,150.13,147.00,140.07,139.57,137.30,125.43,125.28,125.22,124.66,122.63,92.11,56.28,54.44,51.69,45.06,38.16；IR ν _max (KBr)cm ^-1 :3267,3014,2962,2933,1625,1358,1141,899,770。

example 4

The same procedure as in example 1 was repeated except for replacing 2-diethylaminoethylamine in example 1 with an equivalent amount of 1- (2-methoxyethyl) piperazine to give compound 4 as a pale yellow oil: 2- (4- (2-methoxyethyl) piperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 81mg, yield 68.6%, structural characterization data: HRMS (C) ₂₀ H ₂₂ N ₆ OS)m/z[M+H] ⁺ 395.1650 (calculated 395.1649); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.69(s,1H),8.80(d,J＝8.2Hz,1H),7.98(d,J＝1.9Hz,1H),7.92(d,J＝8.0Hz,1H),7.50(t,J＝7.6Hz,1H),7.43(t,J＝7.6Hz,1H),6.10(d,J＝1.9Hz,1H),4.03(br s,4H),3.56(t,J＝5.4Hz,2H),3.38(s,3H),2.69–2.55(m,6H)； ¹³ C NMR(101MHz,CDCl ₃ )δ(ppm):155.72,152.43,149.71,147.24,139.97,139.62,137.39,125.28,125.22,125.13,125.05,122.70,91.59,70.06,58.99,58.03,53.44,44.26；IR ν _max (KBr)cm ^-1 :3065,2968,1610,1447,1358,1236,870,772。

example 5

The same procedure as in example 1 was repeated except for replacing 2-diethylaminoethylamine in example 1 with an equivalent amount of N-methylpiperazine to give a pale yellow solid, compound 5:2- (4-methylpiperazino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 81mg, yield 77.1%, structural characterization data: m.p.112.8-113.7 ℃; HRMS (C) ₁₈ H ₁₈ N ₆ S)m/z[M+H] ⁺ 351.1390 (calculated 351.1386); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.69(s,1H),8.82(d,J＝8.2Hz,1H),8.00(d,J＝2.0Hz,1H),7.96(d,J＝8.0Hz,1H),7.54–7.52(m,1H),7.47–7.45(m,1H),6.12(d,J＝2.0Hz,1H),4.03(br s,4H),2.63–2.49(m,4H),2.37(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):155.72,152.39,149.68,147.22,140.02,139.60,137.37,125.25,125.19,125.11,125.04,122.69,91.62,54.91,46.22,44.34；IR ν _max (KBr)cm ^-1 :3106,3061,2886,1618,1545,1512,1447,1301,1243,991,723。

example 6

The same procedure as in example 1 was repeated except for replacing 2-diethylaminoethylamine in example 1 with an equivalent amount of piperazine to give a pale yellow solid, compound 6: 2-piperazinyl-4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 83mg, yield 82.2%, structural characterization data: m.p.143.3-145.2 deg.C; HRMS (C) ₁₇ H ₁₆ N ₆ S)m/z[M+H] ⁺ 337.1234 (calculated 337.1230); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.69(s,1H),8.81(d,J＝8.2Hz,1H),8.00(d,J＝2.0Hz,1H),7.95(d,J＝8.0Hz,1H),7.52(t,J＝7.6Hz,1H),7.45(t,J＝7.6Hz,1H),6.12(d,J＝2.0Hz,1H),3.98(br s,4H),3.05–2.97(m,4H),1.81(s,1H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):155.84,152.45,149.71,147.25,139.97,139.64,137.40,125.27,125.25,125.13,125.04,122.71,91.58,46.08,45.61；IR ν _max (KBr)cm ^-1 :3456,3102,2887,1538,1530,1444,1305,1245,873,727。

example 7

Weighing 2-piperazinyl-4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 81mg (0.24 mmol), triethylamine 29mg (0.29 mmol) in a round bottom flask, dichloromethane 10mL, acryloyl chloride 22mg (0.24 mmol) slowly added dropwise under ice bath conditions, then stirred at ambient temperature for 1h. After the reaction was completed, the solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (methanol: dichloromethane =1, 35,v/V) to obtain compound 7 as a pale yellow solid: 2- (4-acryloylpiperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 66mg, yield 70.2%, structural characterization data: m.p.147.4-148.8 deg.C; HRMS (C) ₂₀ H ₁₈ N ₆ OS)m/z[M+H] ⁺ 391.1333 (calculated 391.1336); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.74(s,1H),8.79(d,J＝8.2Hz,1H),8.03(d,J＝2.0Hz,1H),7.95(d,J＝8.0Hz,1H),7.52(t,J＝7.6Hz,1H),7.46(t,J＝7.6Hz,1H),6.63(dd,J＝16.8,10.6Hz,1H),6.37(dd,J＝16.8,1.4Hz,1H),6.15(d,J＝2.0Hz,1H),5.77(dd,J＝10.6,1.4Hz,1H),4.04(br s,4H),3.86(br s,2H),3.72(br s,2H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):165.68,155.63,152.17,149.94,147.44,140.46,139.64,137.31,128.51,127.29,125.37,125.25,125.00,124.91,122.80,92.00,45.58,44.50,44.23,41.81；IR ν _max (KBr)cm ^-1 :3463,3024,2886,1731,1609,1439,1244,911,732。

example 8

Weighing 2- (4-methylpiperazino) -4- (benzothiophen-3-yl) pyrazolo [1,5-a][1,3,5]Putting triazine 77mg (0.22 mmol) and p-toluenesulfonic acid monohydrate 42mg (0.22 mmol) in a round-bottom flask, adding methanol 5mL, heating and refluxing at 70 ℃ for 1h, cooling and crystallizing to separate out a light yellow solid, performing suction filtration, and drying to obtain a light yellow solid compound 8:2- (4-methylpiperazine)Yl) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]76mg of triazine p-toluenesulfonate with a yield of 66.1%, and structural characterization data as follows: m.p.147.4-148.8 deg.C; ¹ H NMR(600MHz,DMSO-d6)δ(ppm):9.81(s,1H),9.74(s,1H),8.71(d,J＝8.2Hz,1H),8.24(s,1H),8.20(d,J＝8.0Hz,1H),7.59(t,J＝7.5Hz,1H),7.54(t,J＝7.5Hz,1H),7.49(d,J＝7.4Hz,2H),7.11(d,J＝7.4Hz,2H),6.28(s,1H),3.58–3.12(m,8H),2.86(s,3H),2.28(s,3H)； ¹³ C NMR(151MHz,DMSO-d6)δ(ppm):155.48,151.88,150.09,148.29,146.22,141.78,139.55,138.08,137.27,128.52,126.14,125.97,125.83,125.08,124.51,123.70,92.45,52.46,42.90,41.91,21.24；IR ν _max (KBr)cm ^-1 :3016,2964,2834,1616,1358,1158,761,675,562。

example 9

The same procedure as in example 1 was repeated except for replacing 2-diethylaminoethylamine in example 1 with an equivalent amount of 2-pyrrolidinoethylamine to give a pale yellow solid compound 9:2- (2-pyrrolidinoethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 92mg, yield 84.4%, structural characterization data: m.p.123.7-124.6 ℃; HRMS (C) ₁₉ H ₂₀ N ₆ S)m/z[M+H] ⁺ 365.1538 (calculated 365.1543); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.69(s,1H),8.90(d,J＝7.9Hz,1H),7.95(s,1H),7.87(d,J＝7.8Hz,1H),7.45(t,J＝7.4Hz,1H),7.39(t,J＝7.4Hz,1H),6.23(s,1H),6.11(s,1H),3.65(br s,2H),2.81(br s,2H),2.63(br s,4H),1.80(br s,4H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.27,152.29,150.05,146.83,140.04,139.51,137.34,125.66,125.33,125.19,124.68,122.52,92.00,54.63,54.04,40.01,23.54；IR ν _max (KBr)cm ^-1 :3268,3001,2911,1626,1577,1366,1252,902,731。

example 10

Weighing 140mg (1.2 mmol) of 2-diethylaminoethanol and 28mg (1.2 mmol) of sodium metal into a round-bottomed flask, adding 5mL of dioxane, stirring at room temperature for 1h, and slowly adding 2-methanesulfonyl-4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 100mg (0.3 mmol) was heated under reflux at 110 ℃ for 4h. After completion of the reaction, the reaction mixture was cooled to room temperature, 10mL of ethanol was added to quench the reaction, and the solvent was evaporated under reduced pressure and separated by silica gel column chromatography (methanol: dichloromethane =1, 40, V/V) to obtain a pale yellow solid compound 10:2- (2-diethylaminoethoxy) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 74mg, yield 67.2%, structural characterization data: m.p.61.4-62.8 deg.C, HRMS (C) ₁₉ H ₂₁ N ₅ OS)m/z[M+H] ⁺ 368.1537 (calculated 368.1540); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.90(s,1H),9.05(d,J＝8.2Hz,1H),8.07(d,J＝1.7Hz,1H),7.85(d,J＝7.9Hz,1H),7.47(t,J＝7.6Hz,1H),7.38(t,J＝7.6Hz,1H),6.32(d,J＝1.7Hz,1H),4.53(t,J＝6.4Hz,2H),2.93(t,J＝6.4Hz,2H),2.63(q,J＝7.0Hz,4H),1.04(t,J＝7.0Hz,6H). ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):159.93,151.72,151.37,147.40,141.69,139.34,137.32,126.02,125.69,125.38,124.41,122.47,94.29,66.37,51.07,47.84,11.89；IR ν _max (KBr)cm ^-1 :3105,2902,1634,1577,1366,1243,903,732。

example 11

Weighing 2-methylsulfonyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]164mg (0.5 mmol) of triazine and 268mg (2.0 mmol) of 2-diethylaminoethylamine are placed in a round-bottomed flask, 10mL of dioxane are added, and heating and refluxing are carried out at 110 ℃ for 4 hours. The solvent was evaporated under reduced pressure and separated by silica gel column chromatography (methanol: dichloromethane =1 25,v/V) to give compound 11 as a pale yellow solid: 2- (2-diethylaminoethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 144mg, yield 79.1%, structural characterization data: m.p.151.6-152.5 ℃; HRMS (C) ₂₀ H ₂₅ N ₇ )m/z[M+H] ⁺ 364.2242 (calculated 364.2244); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.26(s,1H),8.81(s,1H),7.97(s,1H),7.36–7.29(m,3H),6.08(s,1H),6.02(s,1H),3.95(s,3H),3.74(br s,2H),2.98(br s,2H),2.82(br s,4H),1.91(br s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.99,151.99,151.37,146.03,139.73,137.21,127.44,123.35,123.13,122.30,109.91,105.18,91.39,67.14,57.23,53.45,37.99,33.75；IR ν _max (KBr)cm ^-1 :3499,3016,2964,1608,1538,1358,1167,761,675,562。

example 12

The same procedure as in example 11 was repeated except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of 2-morpholinoethylamine to give a beige solid compound 12:2- (2-Morpholinylethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 158mg, yield 83.5%, structural characterization data: m.p.187.4-188.9 ℃; HRMS (C) ₂₀ H ₂₅ N ₇ O)m/z[M+H] ⁺ 378.2035 (calculated 378.2037); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.26(s,1H),8.81(s,1H),7.97(s,1H),7.49–7.38(m,3H),6.08(s,1H),6.02(s,1H),3.95(s,3H),3.74(br s,2H),2.98(br s,2H),2.82(br s,4H),1.91(br s,4H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.99,151.99,151.37,146.03,139.73,137.21,127.44,123.35,123.13,122.30,109.91,105.18,91.39,67.14,57.23,53.45,37.99,33.75；IR ν _max (KBr)cm ^-1 :3482,2921,2868,1625,1538,1435,1374,769,622。

example 13

The same procedures as in example 11 were repeated except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of 1- (2-methoxyethyl) piperazine to give a reaction productTo beige compound 13:2- (4- (2-methoxyethyl) piperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 137mg, yield 69.8%, structural characterization data: m.p.119.3-120.4 ℃; HRMS (C) ₂₁ H ₂₅ N ₇ O)m/z[M+H] ⁺ 392.2189 (calculated 392.2193); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.31(s,1H),8.67(d,J＝5.6Hz,1H),8.00(s,1H),7.47(d,J＝7.1Hz,1H),7.42–7.40(m,2H),6.10(s,1H),4.12(br s,4H),3.97(s,3H),3.64(br s,2H),3.43(s,3H),2.72(br s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.41,152.11,150.95,146.26,139.86,137.23,127.36,123.31,122.71,122.35,110.01,105.70,91.11,69.95,59.04,58.08,53.51,44.14,33.79；IR ν _max (KBr)cm ^-1 :2968,2911,2853,1627,1545,1479,1382,1114,854,764,626。

example 14

The same procedure as in example 11 was repeated except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of N-methylpiperazine to give a beige solid compound 14:2- (4-methylpiperazino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 130mg, yield 74.7%, structural characterization data: m.p.221.5-223.1 ℃; HRMS (C) ₁₉ H ₂₁ N ₇ )m/z[M+H] ⁺ 348.1924 (calculated 348.1931); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.23(s,1H),8.63–8.50(m,1H),7.91(d,J＝2.0Hz,1H),7.42–7.35(m,1H),7.35–7.30(m,2H),6.01(d,J＝2.0Hz,1H),4.04(br s,4H),3.89(s,3H),2.58(br s,4H),2.36(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.47,152.09,150.89,146.21,139.85,137.20,127.35,123.27,122.69,122.31,109.97,105.69,91.09,55.02,46.28,44.34,33.73；IR ν _max (KBr)cm ^-1 :2918,2862,1627,1528,1415,1374,1114,862,772,626。

example 15

The same procedure as in example 11 was repeated except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of piperazine to give a beige solid compound 15: 2-piperazinyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 112mg, yield 67.1%, structural characterization data: m.p.173.4-175.0 deg.C; HRMS (C) ₁₈ H ₁₉ N ₇ )m/z[M+H] ⁺ 334.1772 (calculated 334.1775); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.25(s,1H),8.62(d,J＝7.2Hz,1H),7.96(s,1H),7.44–7.32(m,3H),6.07(s,1H),3.99(br s,4H),3.89(s,3H),3.02(br s,4H),1.84(s,1H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.56,152.12,150.86,146.19,139.82,137.19,127.35,123.25,122.69,122.29,109.95,105.71,91.03,46.14,45.60,33.71；IR ν _max (KBr)cm ^-1 :3456,3105,2911,2902,1642,1569,1366,1252,903,732。

example 16

With equal amount of 2- (piperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine substitution of 2- (piperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a in example 7][1,3,5]Triazine, the other steps being the same as in example 7, gave compound 16 as a pale yellow solid: 2- (4-acryloylpiperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 48mg, yield 51.6%, structural characterization data: m.p.219.9-221.5 ℃; HRMS (C) ₂₁ H ₂₁ N ₇ O)m/z[M+H] ⁺ 388.1878 (calculated 388.1880); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.31(s,1H),8.70–8.55(m,1H),8.00(d,J＝1.9Hz,1H),7.52–7.42(m,1H),7.42–7.36(m,2H),6.66(dd,J＝16.8,10.6Hz,1H),6.38(dd,J＝16.8,1.4Hz,1H),6.11(s,1H),5.77(dd,J＝10.6,1.4Hz,1H),4.08(br s,4H),3.96(s,3H),3.88(br s,2H),3.75(br s,2H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):165.66,156.29,151.81,151.12,146.38,140.12,137.26,128.31,127.43,127.32,123.41,122.59,122.46,110.09,105.56,91.39,45.67,44.51,44.19,41.87,33.79；IR ν _max (KBr)cm ^-1 :3064,2903,1780,1634,1528,1431,1243,1211,886,739。

example 17

The same procedure as in example 11 was followed, except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of 2-pyrrolidinoethylamine, to give a beige solid compound 17:2- (2-pyrrolidinoethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]141mg of triazine, with a yield of 77.9%, and the structural characterization data: m.p.181.3-182.1 deg.C; HRMS (C) ₂₀ H ₂₃ N ₇ )m/z[M+H] ⁺ 362.2081 (calculated 362.2088); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.23(s,1H),8.80(d,J＝6.1Hz,1H),7.95(d,J＝1.6Hz,1H),7.47–7.37(m,1H),7.38–7.35(m,2H),6.07(s,1H),6.01(s,1H),3.91(s,3H),3.75(br s,2H),2.95(br s,2H),2.79(br s,4H),1.88(br s,4H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):157.11,152.07,151.32,145.94,139.69,137.18,127.49,123.29,123.26,122.25,109.81,105.34,100.01,54.86,53.99,40.45,33.70,23.57；IR ν _max (KBr)cm ^-1 :3457,2972,2852,1607,1530,1427,1116,882,744,622。

example 18

The same procedure as in example 11 was repeated except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of diethylamine to give a pale yellow solid compound 18: 2-diethylamino-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 130mg, yield 81.2%, structural characterization data: m.p.134.1-134.6 deg.C; HRMS (C) ₁₈ H ₂₀ N ₆ )m/z[M+H] ⁺ 321.1817 (calculated 321.1822); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.28(s,1H),8.72(d,J＝8.7Hz,1H),7.96(s,1H),7.40(s,1H),7.38–7.34(m,2H),6.07(s,1H),3.89(br s,4H),3.82(s,3H),1.35(br s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.00,152.58,150.60,145.92,139.62,137.16,127.53,123.13,122.76,122.09,109.84,105.95,90.53,42.38,33.65,13.52；IR ν _max (KBr)cm ^-1 :2972,2852,1608,1530,1434,1383,1106,890,744,631。

example 19

The same procedure as in example 11 was repeated except for replacing 2-diethylaminoethylamine in example 11 with an equivalent amount of N, N' -trimethylethylenediamine to give a pale yellow solid compound 19:2- (N, N, N' -trimethylethylenediamine) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 132mg, yield 75.4%, structural characterization data: m.p.129.8-130.3 ℃; HRMS (C) ₁₉ H ₂₃ N ₇ )m/z[M+H] ⁺ 350.2084 (calculated 350.2088); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.15(s,1H),8.66(t,J＝6.9Hz,1H),7.91(d,J＝2.1Hz,1H),7.44–7.09(m,3H),6.03(s,1H),3.89(br s,2H),3.76(s,3H),3.28(s,3H),2.62(d,J＝6.1Hz,2H),2.33(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):156.02,152.07,151.05,146.31,139.94,137.23,127.35,123.31,122.54,122.35,110.03,105.67,91.17,47.23,33.76,27.10；IR ν _max (KBr)cm ^-1 :3057,2943,2894,1618,1530,1472,1236,886,739。

example 20

1. In a round-bottomed flask, 2.0g (12.8 mmol) of 4-fluoro-3-nitroaniline, 1.9g (19.1 mmol) of N-methylpiperazine and 2.5g (19.1 mmol) of N, N-Diisopropylethylamine (DIEA) were weighed, 25mL of acetonitrile was added, and the mixture was reacted at reflux temperature for 12 hours. Residue after evaporation of solventThe material was subjected to silica gel column chromatography (methanol: dichloromethane =1, 20,v/V) to give 4- (4-methylpiperazinyl) -3-nitroaniline 2.6g as an orange-red solid in 86.6% yield and with the structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):7.08(d,J＝8.3Hz,1H),7.00(d,J＝2.2Hz,1H),6.81(dd,J＝8.3,2.2Hz,1H),3.73(s,2H),2.97(br s,4H),2.55(br s,4H),2.34(s,3H)。

2. 507mg (1.7 mmol) of 1-methylindole-3-formyl-pyrazol-5-ylthiourea, 400mg (1.7 mmol) of 4- (4-methylpiperazinyl) -3-nitroaniline, 459mg (1.7 mmol) of mercuric chloride and 325mg (3.4 mmol) of triethylamine are weighed into a round-bottomed flask, 10ml of DMF is added, and the mixture is reacted at 130 ℃ for 12 hours in a nitrogen atmosphere. After completion of the reaction, the reaction mixture was cooled to room temperature, insoluble matter was removed by suction filtration, the filtrate was diluted with ethyl acetate and washed with a saturated aqueous sodium chloride solution 3 times, the organic phase was collected, dried over anhydrous sodium sulfate, concentrated by filtration, and separated by silica gel column chromatography (methanol: dichloromethane =1, 200, V/V) to give 2- (4- (4-methylpiperazinyl) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a) as an orange-red solid][1,3,5]Triazine 263mg, yield 32.3%, structural characterization data are: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.24(s,1H),8.68(d,J＝7.6Hz,1H),8.28(d,J＝2.4Hz,1H),8.02(d,J＝2.0Hz,1H),7.75(dd,J＝8.8,2.4Hz,1H),7.43–7.29(m,4H),7.19(d,J＝8.8Hz,1H),6.22(d,J＝2.0Hz,1H),3.90(s,3H),3.16–3.01(m,4H),2.60(d,J＝3.9Hz,4H),2.37(s,3H)。

3. weighing 2- (4- (4-methylpiperazino) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 200mg (0.41 mmol), iron powder 140mg (2.5 mmol), ammonium chloride 16mg (0.29 mmol) in a round-bottomed flask, 4mL of ethanol, 8mL of tetrahydrofuran and 4mL of water were added, and the mixture was heated under reflux at 80 ℃ for 4 hours. After the reaction, the reaction mixture was cooled to room temperature, insoluble matter was removed by suction filtration, the filtrate was evaporated under reduced pressure to remove the solvent, the residue was dissolved in methylene chloride, and the mixture was washed with a saturated aqueous sodium bicarbonate solution and a saturated aqueous sodium chloride solution 2 times, and the organic phase was collected, dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography (methanol: methylene chloride =1, 200V/V) to give 2- (4- (4-methylpiperazinyl) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 106mg, yield 57.0%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.30(s,1H),8.79(s,1H),8.02(s,1H),7.44–7.36(m,4H),7.06–7.04(m,2H),6.95(s,1H),6.20(s,1H),4.12(s,2H),3.96(s,3H),2.98(br s,4H),2.64(br s,4H),2.39(s,3H)。

4. weighing 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]106mg (0.23 mmol) of triazine and 36mg (0.35 mmol) of triethylamine are placed in a round-bottom flask, 15mL of dichloromethane solution is added, 25mg (0.28 mmol) of acryloyl chloride is slowly added under ice bath conditions, and stirring is carried out at room temperature for 1h. After the reaction was complete, 15mL of saturated aqueous sodium bicarbonate was added to quench the reaction, and the solution was separated. The organic phase was washed with a saturated aqueous sodium chloride solution, the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was subjected to silica gel column chromatography (methanol: dichloromethane =1, 40, V/V) to obtain compound 20:2- (3-Acrylamido-4- (4-methylpiperazino) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 52mg, yield 44.4%, structural characterization data m.p.137.1-138.7 ℃; HRMS (C) ₂₈ H ₂₉ N ₉ O)m/z[M+H] ⁺ 508.2565 (calculated 508.2568); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.22(s,1H),8.86(s,1H),8.76(dd,J＝5.8,2.8Hz,1H),8.51(s,1H),8.00(br s,2H),7.46–7.30(m,4H),7.25(d,J＝8.6Hz,1H),6.41(d,J＝16.9Hz,1H),6.31(dd,J＝16.9,10.2Hz,1H),6.22(d,J＝1.8Hz,1H),5.79(d,J＝10.2Hz,1H),3.89(s,3H),2.93(br s,4H),2.62(s,3H),2.40(br s,4H). ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.32,154.25,151.34,151.19,146.06,139.85,137.17,136.78,136.15,133.74,132.01,127.34,126.99,123.47,123.41,122.38,121.35,115.26,110.73,109.80,105.07,92.76,56.06,52.48,46.15,33.72；IR ν _max (KBr)cm ^-1 :3553,3380,3024,2932,1670,1587,1514,1459,1377,1075,864,745。

example 21

1. In a round-bottomed flask, 0.78g (5.0 mmol) of 4-fluoro-3-nitroaniline, 1.80g (25.0 mmol) of diethylamine, and 1.94g (15 mmol) of N, N-Diisopropylethylamine (DIEA) were weighed, and 25mL of acetonitrile was added to react at 56 ℃ for 12 hours. The solvent was evaporated and the residue was subjected to silica gel column chromatography (dichloromethane) to give 528mg of 4-diethylamino-3-nitroaniline as a reddish brown solid in a yield of 50.5%, according to the structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):7.09(d,J＝8.7Hz,1H),7.01(d,J＝2.6Hz,1H),6.83(dd,J＝8.7,2.6Hz,1H),3.81–3.79(m,4H),2.95–2.92(m,6H)。

2. the same procedures used in example 20, step 2, were repeated except for replacing 4- (4-methylpiperazino) -3-nitroaniline in example 20, step 2, with an equivalent amount of 4- (diethylamino) -3-nitroaniline to give the orange-red body 2- (4-diethylamino-3-nitroanilino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 399mg, yield 51.4%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.20(s,1H),8.64(d,J＝7.6Hz,1H),8.16(s,1H),8.01(d,J＝1.6Hz,1H),7.66(dd,J＝8.8,1.9Hz,1H),7.41(s,1H),7.36–7.27(m,3H),7.20(d,J＝8.8Hz,1H),6.20(d,J＝1.6Hz,1H),3.86(s,3H),3.10(q,J＝7.1Hz,4H),1.08(t,J＝7.1Hz,6H)。

3. with equal mass of 2- (4-diethylamino-3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, the other steps were the same as in step 3 of example 20, to give 2- (4-diethylamino-3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 128mg, yield 73.5%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.29(s,1H),8.78(d,J＝7.7Hz,1H),8.02(d,J＝2.0Hz,1H),7.43(d,J＝8.0Hz,1H),7.40–7.30(m,3H),7.11–6.99(m,2H),6.96(dd,J＝8.3,2.1Hz,1H),6.20(d,J＝2.0Hz,1H),4.22(s,2H),3.95(s,3H),2.96(q,J＝7.1Hz,4H),1.03(t,J＝7.1Hz,6H)。

4. with equal mass of 2- (4-diethylamino-3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine substituted triazineExample 20 2- (4- (4-Methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give compound 21 as a yellow solid: 2- (3-Acrylamido-4-diethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 52mg, yield 46.8%, structural characterization data: m.p.109.1-110.8 ℃; HRMS (C) ₂₇ H ₂₈ N ₈ O)m/z[M+H] ⁺ 481.2461 (calculated 481.2495); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.20(s,1H),9.16(d,J＝17.2Hz,1H),8.72(s,1H),8.57(s,1H),8.06(d,J＝7.9Hz,1H),7.99(s,1H),7.38–7.31(m,4H),7.22(d,J＝8.6Hz,1H),6.41(d,J＝16.9Hz,1H),6.31(dd,J＝16.9,10.1Hz,1H),6.22(s,1H),5.75(d,J＝10.1Hz,1H),3.85–3.78(m,3H),2.94(q,J＝7.1Hz,4H),0.98(t,J＝7.1Hz,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.33,154.24,151.26,151.20,145.99,139.82,137.12,136.97,136.77,133.77,132.10,127.33,126.82,123.47,123.41,123.34,122.31,114.87,109.97,109.74,105.05,92.71,50.07,33.64,13.09；IR ν _max (KBr)cm ^-1 :3298,2968,2923,1670,1596,1514,1459,1367,1230,1158,1065,965,864,773,736。

example 22

1. The same procedure as in step 1 of example 20 was repeated except for replacing N-methylpiperazine in step 1 of example 20 with an equal amount of morpholine to give 2.6g of 4-morpholinyl-3-nitroaniline as an orange-red solid in a yield of 91.0%, and the structural characterization data are as follows: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):7.09(d,J＝8.7Hz,1H),7.01(d,J＝2.6Hz,1H),6.83(dd,J＝8.7,2.6Hz,1H),3.81–3.79(m,4H),2.95–2.92(m,4H)。

2. the procedure of example 20 was repeated except for replacing 4- (4-methylpiperazino) -3-nitroaniline in example 20, step 2, with an equivalent amount of 4-morpholinyl-3-nitroaniline to give 2- (3-nitro-4-morpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo-orange-red isomer[1,5-a][1,3,5]Triazine 219mg, yield 27.2%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.35(s,1H),8.76(d,J＝7.5Hz,1H),8.41(d,J＝2.6Hz,1H),8.09(d,J＝2.1Hz,1H),7.84(dd,J＝8.8,2.6Hz,1H),7.47(d,J＝7.5Hz,1H),7.43–7.36(m,2H),7.29(s,1H),7.26(d,J＝8.8Hz,1H),6.29(d,J＝2.1Hz,1H),3.99(s,3H),3.92–3.85(m,4H),3.15–3.02(m,4H)。

3. with equal amount of 2- (3-nitro-4-morpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine As an alternative to 2- (4- (4-Methylpiperazino) -3-Nitrophenylamino) -4- (1-Methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, the other steps were the same as in step 3 of example 20, to give 2- (3-amino-4-morpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 121mg, yield 67.0%, structural characterization data: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):9.51(s,1H),9.35(s,1H),9.00(d,J＝5.1Hz,1H),8.13(d,J＝2.1Hz,1H),7.65(d,J＝8.1Hz,1H),7.41–7.37(m,1H),7.34(t,J＝7.4Hz,1H),7.27(s,1H),7.06(d,J＝7.6Hz,1H),6.92(d,J＝8.5Hz,1H),6.17(d,J＝2.1Hz,1H),4.88(s,2H),4.02(s,3H),3.84–3.62(m,4H),2.96–2.70(m,4H)。

4. with equal amount of 2- (3-amino-4-morpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give a yellow solid compound 22:2- (3-Acryloylamino-4-morpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 51mg, yield 44.7%, structural characterization data m.p.169.3-171.3 ℃; HRMS (C) ₂₇ H ₂₆ N ₈ O ₂ )m/z[M+H] ⁺ 495.2258 (calculated 495.2251); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.27(s,1H),8.85(s,1H),8.83–8.74(m,1H),8.52(s,1H),8.05(d,J＝7.9Hz,1H),8.02(d,J＝1.4Hz,1H),7.50–7.37(m,3H),7.30(s,1H),7.25(d,J＝7.1Hz,1H),6.42(d,J＝16.9Hz,1H),6.31(dd,J＝16.9,10.2Hz,1H),6.24(d,J＝1.4Hz,1H),5.81(d,J＝10.2Hz,1H),3.94(s,3H),3.89(s,4H),2.90(s,4H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.31,154.21,151.34,151.16,146.06,139.87,137.17,137.05,135.85,133.76,131.91,127.35,127.15,123.46,123.40,122.37,121.38,115.19,110.80,109.81,105.06,92.80,67.77,52.94,33.71；IR ν _max (KBr)cm ^-1 :3289,2976,2940,1670,1596,1523,1359,1230,1157,911,764,736。

example 23

1. The same procedure as in step 1 of example 20 was repeated except for replacing N-methylpiperazine in step 1 of example 20 with thiomorpholine in an equivalent amount to give 2.0g of 4-thiomorpholinyl-3-nitroaniline as an orange-red solid in a yield of 65.3%, and the structural characterization data were as follows: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):7.17(d,J＝8.7Hz,1H),6.85(d,J＝2.7Hz,1H),6.78(dd,J＝8.7,2.7Hz,1H),5.49(s,2H),3.08–2.95(m,4H),2.70–2.59(m,4H)。

2. the same procedures used in example 20, step 2, were repeated except for replacing 4- (4-methylpiperazino) -3-nitroaniline in example 20, step 2, with an equivalent amount of 4-thiomorpholino-3-nitroaniline to give 2- (3-nitro-4-thiomorpholino phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine 249mg as an orange-red body in 30.2% yield.

3. With equal mass of 2- (3-nitro-4-thiomorpholino phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, and the other steps were carried out in the same manner as in step 3 of example 20 to give 2- (3-amino-4-thiomorpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 112mg, yield 61.5%, structural characterization data: ¹ H NMR(600MHz,DMSO-d6)δ(ppm):9.49(s,1H),9.35(s,1H),9.00(s,1H),8.13(d,J＝2.0Hz,1H),7.65(d,J＝8.1Hz,1H),7.39(t,J＝7.2Hz,1H),7.34(t,J＝7.4Hz,1H),7.25(s,1H),7.05(d,J＝7.5Hz,1H),6.91(d,J＝8.4Hz,1H),6.17(d,J＝2.0Hz,1H),4.87(s,2H),4.02(s,3H),3.02(br s,4H),2.80(br s,4H)。

4. with an equivalent amount of 2- (3-amino-4-thiomorpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in example 20, step 4, to give compound 23 as a beige solid: 2- (3-Acryloylamino-4-thiomorpholinylphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 63mg, yield 53.9%, structural characterization data: m.p.221.4-223.4 ℃; HRMS (C) ₂₇ H ₂₆ N ₈ OS)m/z[M+H] ⁺ 511.2022 (calculated 511.2023); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.22(s,1H),8.76–8.75(m,2H),8.49(s,1H),8.03(d,J＝8.2Hz,1H),8.01(d,J＝1.9Hz,1H),7.40–7.35(m,3H),7.34(s,1H),7.20(d,J＝8.6Hz,1H),6.42(d,J＝16.9Hz,1H),6.29(dd,J＝16.9,10.1Hz,1H),6.23(d,J＝1.9Hz,1H),5.80(d,J＝10.1Hz,1H),3.89(s,3H),3.20–3.02(m,4H),2.83(br s,4H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.28,154.19,151.34,151.15,146.06,139.88,137.18,137.05,133.56,131.90,127.35,127.18,123.46,123.41,122.39,121.83,115.09,115.07,110.67,109.81,105.06,92.82,54.81,33.73,29.71；IR ν _max (KBr)cm ^-1 :3453,3035,2901,1660,1605,1523,1447,1367,1312,965,882,773,736。

example 24

1. The same procedures used in step 1 of example 20 were repeated except for replacing N-methylpiperazine in step 1 of example 20 with an equivalent amount of N, N, N '-trimethylethylenediamine to give 2.6g of 4- (N, N, N' -trimethylethylenediamine) -3-nitroaniline as a reddish brown solid in a yield of 84.6 g8%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):7.08(d,J＝8.7Hz,1H),6.95(d,J＝2.8Hz,1H),6.79(dd,J＝8.7,2.8Hz,1H),3.71(s,2H),3.09–2.99(m,2H),2.72(s,3H),2.46–2.35(m,2H),2.20(s,6H)。

2. the 4- (4-methylpiperazino) -3-nitroaniline in step 2 in example 20 was replaced with an equivalent amount of 4- (N, N, N '-trimethylethylenediamine) -3-nitroaniline and the other steps were the same as in step 2 in example 20 to give an orange-red body of 2- (3-nitro-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 318mg, its rate is 38.5%, the structural characterization data are: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.29(s,1H),8.72(d,J＝7.5Hz,1H),8.28(s,1H),8.04(d,J＝1.8Hz,1H),7.73(d,J＝8.0Hz,1H),7.43(d,J＝7.9Hz,1H),7.40–7.31(m,3H),7.21(d,J＝8.8Hz,1H),6.24(d,J＝1.8Hz,1H),3.94(s,3H),3.32–3.24(m,2H),2.85(s,3H),2.64–2.58(m,2H),2.33(s,6H)。

3. with an equal mass of 2- (3-nitro-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine As an alternative to 2- (4- (4-Methylpiperazino) -3-Nitrophenylamino) -4- (1-Methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, the other steps were the same as in step 3 of example 20, to give 2- (3-amino-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 105mg, yield 56.4%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.29(s,1H),8.83–8.75(m,1H),8.02(d,J＝2.1Hz,1H),7.44(d,J＝7.2Hz,1H),7.40–7.35(m,2H),7.31(d,J＝2.2Hz,1H),7.05–7.03(m,2H),6.93(dd,J＝8.4,2.2Hz,1H),6.20(d,J＝2.1Hz,1H),3.95(s,3H),3.05(t,J＝6.5Hz,2H),2.70(s,3H),2.59–2.53(m,2H),2.38(br s,6H)。

4. with an equal mass of 2- (3-amino-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine As an alternative to 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in the fourth step of example 20][1,3,5]Triazine, further procedure and example 20The same four steps gave compound 24 as a yellow solid: 2- (3-Acryloylamino-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 59mg, its yield is 50.5%. The structural characterization data are: m.p.121.3-123.3 ℃; HRMS (C) ₂₈ H ₃₁ N ₉ O)m/z[M+H] ⁺ 510.2725 (calculated 510.2724); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.96(s,1H),9.21(s,1H),8.76(s,1H),8.58(s,1H),8.02–7.90(m,2H),7.43(s,1H),7.36(s,3H),7.22(s,1H),6.67–6.53(m,1H),6.46(d,J＝16.2Hz,1H),6.21(s,1H),5.73(d,J＝7.4Hz,1H),3.87(s,3H),2.98(br s,2H),2.67(s,3H),2.57(br s,2H),2.40(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):164.22,154.29,151.34,151.17,146.04,139.86,137.17,136.94,136.86,135.70,132.04,127.35,126.97,123.48,123.41,122.47,122.40,115.28,111.55,109.79,105.07,92.68,77.29,77.08,76.87,56.38,54.91,44.54,43.42,33.71；IR ν _max (KBr)cm ^-1 :3474,2986,2932,1670,1597,1459,1358,974,874,773。

example 25

1. This step is the same as in step 1 of example 22.

2. 845mg (2.8 mmol) of benzothiophene-3-formyl-pyrazol-5-ylthiourea, 624mg (2.8 mmol) of 4-morpholinyl-3-nitroaniline, 759mg (2.8 mmol) of mercuric chloride and 566mg (5.6 mmol) of triethylamine were weighed out and placed in a round-bottomed flask, 15mL of DMF was added, and the mixture was reacted at 130 ℃ for 12 hours under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was cooled to room temperature, insoluble substances were removed by suction filtration, the filtrate was diluted with ethyl acetate and washed with a saturated aqueous sodium chloride solution 3 times, the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was subjected to silica gel column chromatography (methanol: dichloromethane =1:200 to 100, V/V) to give an orange-red body, 2- (3-nitro-4-morpholinophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 742mg, yield 56.0%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.83(s,1H),8.86(d,J＝8.0Hz,1H),8.32(d,J＝2.2Hz,1H),8.09(d,J＝2.1Hz,1H),7.91(d,J＝7.5Hz,1H),7.77(dd,J＝8.8,2.5Hz,1H),7.50(s,1H),7.48–7.42(m,2H),7.18(d,J＝8.8Hz,1H),6.30(d,J＝2.1Hz,1H),3.88–3.82(m,4H),3.07–3.00(m,4H)。

3. with equal mass of 2- (3-nitro-4-morpholinylphenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine As an alternative to 2- (4- (4-Methylpiperazino) -3-Nitrophenylamino) -4- (1-Methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, the other steps were the same as in step 3 of example 20, to give 2- (3-amino-4-morpholinophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 110mg, yield 60.5%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.83(s,1H),8.96(d,J＝8.1Hz,1H),8.07(d,J＝2.1Hz,1H),7.96(d,J＝8.0Hz,1H),7.52(t,J＝7.5Hz,1H),7.47(t,J＝7.4Hz,1H),7.36(d,J＝2.1Hz,1H),7.14(s,1H),7.03(d,J＝8.4Hz,1H),6.97(dd,J＝8.4,2.2Hz,1H),6.28(d,J＝2.1Hz,1H),4.13(s,2H),3.88(br s,4H),3.05–2.83(m,4H)。

4. with equal amount of 2- (3-amino-4-morpholinylphenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give compound 25 as a yellow solid: 2- (3-Acryloylamino-4-morpholinylphenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 93mg, yield 81.3%, structural characterization data: m.p.131.8-132.8 ℃; HRMS (C) ₂₆ H ₂₃ N ₇ O ₂ S)m/z[M+H] ⁺ 498.1710 (calculated 498.1707); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.88(s,1H),9.00(d,J＝8.2Hz,1H),8.84(s,1H),8.54(s,1H),8.07(d,J＝2.0Hz,1H),8.05(d,J＝8.0Hz,1H),7.94(d,J＝8.0Hz,1H),7.55(t,J＝7.6Hz,1H),7.47(t,J＝7.6Hz,1H),7.43(s,1H),7.25(s,1H),6.42(d,J＝16.9Hz,1H),6.34–6.26(m,2H),5.81(d,J＝10.3Hz,1H),3.89(br s,4H),2.90(br s,4H)； ¹³ CNMR(151MHz,CDCl ₃ )δ(ppm):163.38,153.50,151.47,150.05,147.06,140.98,139.46,137.28,136.56,136.18,133.83,131.81,127.32,125.85,125.43,125.30,124.45,122.52,121.48,115.14,110.88,93.47,67.73,52.93；IR ν _max (KBr)cm ^-1 :3482,3102,2947,1674,1592,1512,1358,908,848,770。

example 26

1. This step is the same as step 1 of example 20.

2. The same procedures used in example 25, step 2, were repeated except for replacing 4-morpholinyl-3-nitroaniline in example 25, step 2, with an equivalent amount of 4- (4-methylpiperazinyl) -3-nitroaniline to give 2- (3-nitro-4- (4-methylpiperazinyl) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a) as a orange-red solid][1,3,5]Triazine 843mg, yield 61.8%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.86(s,1H),8.91(d,J＝6.6Hz,1H),8.34(s,1H),8.10(s,1H),7.95(d,J＝7.0Hz,1H),7.80(d,J＝7.7Hz,1H),7.58–7.43(m,2H),7.35(s,1H),7.23(d,J＝8.1Hz,1H),6.33(s,1H),3.10(br s,4H),2.61(br s,4H),2.37(s,3H)。

3. with equal mass of 2- (3-nitro-4- (4-methylpiperazino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine As an alternative to 2- (4- (4-Methylpiperazino) -3-Nitrophenylamino) -4- (1-Methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, the other steps were the same as in step 3 of example 20, to give 2- (3-amino-4- (4-methylpiperazinyl) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a as a tan solid][1,3,5]Triazine 111mg, yield 59.3%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.82(s,1H),8.95(d,J＝8.2Hz,1H),8.06(d,J＝2.0Hz,1H),7.95(d,J＝8.0Hz,1H),7.52(t,J＝7.5Hz,1H),7.47(t,J＝7.4Hz,1H),7.33(d,J＝1.6Hz,1H),7.13(s,1H),7.04(d,J＝8.4Hz,1H),6.96(dd,J＝8.4,1.9Hz,1H),6.27(d,J＝1.9Hz,1H),4.10(s,2H),2.97(br s,4H),2.61(br s,4H),2.38(s,3H).

4. with equal amount of 2- (3-amino-4- (4-methylpiperazino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give compound 26 as a yellow solid: 2- (3-Acrylamido-4- (4-methylpiperazino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 52mg, yield 44.6%, structural characterization data: m.p.136.1-137.2 ℃; HRMS (C) ₂₇ H ₂₆ N ₈ OS)m/z[M+H] ⁺ 511.2025 (calculated 511.2023); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.89(s,1H),9.02(d,J＝8.2Hz,1H),8.86(s,1H),8.52(s,1H),8.07(d,J＝2.1Hz,1H),8.04(d,J＝8.0Hz,1H),7.95(d,J＝8.0Hz,1H),7.57(t,J＝7.6Hz,1H),7.49(t,J＝7.2Hz,1H),7.37(s,1H),7.28(d,J＝8.7Hz,1H),6.42(d,J＝17.0Hz,1H),6.37–6.23(m,2H),5.81(d,J＝9.6Hz,1H),2.94(br s,4H),2.64(br s,4H),2.40(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.39,153.56,151.53,150.07,147.06,140.92,139.47,137.30,136.56,136.28,133.86,131.92,127.13,125.86,125.45,125.31,124.49,122.53,121.47,115.17,110.76,93.44,56.07,52.51,46.18；IR ν _max (KBr)cm ^-1 :3394,3107,2964,1640,1568,1373,1142,1093,959,775。

example 27

1. This step is the same as step 1 of example 24.

2. The same procedures used in example 25, step 2, were repeated except for replacing 4-morpholinyl-3-nitroaniline used in example 25, step 2, with an equivalent amount of 4- (N, N, N '-trimethylethylenediamine) -3-nitroaniline to give the orange-red body 2- (3-nitro-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 578mg, yield 42.3%, structural characterizationThe data are as follows: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.86(s,1H),8.92(d,J＝8.2Hz,1H),8.29(d,J＝1.8Hz,1H),8.10(d,J＝2.0Hz,1H),7.96(d,J＝7.9Hz,1H),7.74(dd,J＝8.9,2.4Hz,1H),7.52(t,J＝7.5Hz,1H),7.48(t,J＝7.5Hz,1H),7.27(s,1H),7.21(d,J＝8.9Hz,1H),6.32(d,J＝2.0Hz,1H),3.42–3.13(m,2H),2.87(s,3H),2.67–2.47(m,2H),2.28(s,6H)。

3. the equivalent mass of 2- (3-nitro-4- (N, N, N '-trimethylethylenediamino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine used in step 3 of example 20 was substituted for 2- (4- (4-methylpiperazinyl) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine used in example 20, and the other steps were the same as in step 3 of example 20 to give 129mg of 2- (3-amino-4- (N, N, N' -trimethylethylenediamino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a ] [1,3,5] triazine as a yellow-green solid in 68.4% yield.

4. With an equivalent mass of 2- (3-amino-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give compound 27 as a yellow solid: 2- (3-Acryloylamino-4- (N, N, N' -trimethylethylenediamine) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 80mg, yield 68.4%, structural characterization data: m.p.102.9-103.6 deg.C; HRMS (C) ₂₇ H ₂₈ N ₈ OS)m/z[M+H] ⁺ 513.2167 (calculated 513.2180); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):10.31(s,1H),9.89(s,1H),9.03(d,J＝8.2Hz,1H),8.57(d,J＝2.5Hz,1H),8.07(d,J＝2.1Hz,1H),8.06(s,1H),7.96(d,J＝8.0Hz,1H),7.59(t,J＝7.5Hz,1H),7.49(t,J＝7.1Hz,1H),7.38(s,1H),7.29(d,J＝8.6Hz,1H),6.47(dd,J＝16.9,1.4Hz,1H),6.35(s,1H),6.31(d,J＝2.1Hz,1H),5.74(dd,J＝10.2,1.4Hz,1H),2.89(br s,2H),2.74(s,3H),2.32(br s,2H),2.28(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):164.07,153.57,151.58,150.00,147.00,140.88,139.44,137.47,137.31,136.19,136.07,131.99,126.81,125.91,125.46,125.30,124.50,122.73,122.49,115.03,111.39,93.36,57.35,45.53,43.25,29.71；IR ν _max (KBr)cm ^-1 :3440,3065,2911,1675,1602,1521,1252,878,739。

example 28

1. The same procedures used in example 20, step 2, were repeated except for replacing 4- (4-methylpiperazino) -3-nitroaniline in example 20, step 2, with an equivalent amount of 4-dimethylamino-3-nitroaniline to give the orange-red colored body 2- (3-nitro-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]197mg triazine, 27.0% yield, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.22(s,1H),8.65(d,J＝7.2Hz,1H),8.23(s,1H),8.00(d,J＝1.8Hz,1H),7.69(d,J＝7.6Hz,1H),7.36–7.27(m,4H),7.05(d,J＝8.9Hz,1H),6.19(d,J＝1.8Hz,1H),3.88(s,3H),2.87(s,6H)。

2. with equal mass of 2- (3-nitro-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, and the other steps were the same as in step 3 of example 20, to give 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 71mg, yield 44.6%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.24(s,1H),8.74(d,J＝7.5Hz,1H),8.01(t,J＝4.0Hz,1H),7.39–7.31(m,3H),7.30(d,J＝2.2Hz,1H),7.14(s,1H),7.03(d,J＝8.3Hz,1H),6.94(dd,J＝8.3,2.0Hz,1H),6.18(d,J＝2.0Hz,1H),4.12(s,2H),3.89(s,3H),2.69(s,6H)。

3. weighing 58mg (0.35 mmol) of trans-4-dimethylamino crotonate into a round-bottom flask, adding a catalytic amount of DMF (1 drop) and 4mL of tetrahydrofuran, slowly adding 0.3mL of oxalyl chloride under an ice-bath condition in a nitrogen atmosphere, reacting at ambient temperature for 1.5h after the dropping is finished, and completely dissolving a substrate into dark red to finish the preparation of the acyl chloride. The reaction mixture was evaporated under reduced pressure to remove the solventThen, 4mL of anhydrous tetrahydrofuran containing 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a was added thereto][1,3,5]Triazine 71mg (0.18 mmol) of N-methylpyrrolidone (NMP) solution (3 mL) was stirred at room temperature for 3h. The reaction mixture was adjusted to pH about 9 with 2mol/L aqueous sodium hydroxide solution, extracted with 3 × 15mL of ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated by filtration, and the residue was subjected to silica gel column chromatography (methanol: dichloromethane =1, 20, V/V) to give compound 28 as a yellow-green solid: 2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 12mg, yield 13.6%, structural characterization data: m.p.134.7-136.3 ℃; HRMS (C) ₂₈ H ₃₁ N ₉ O)m/z[M+H] ⁺ 510.2729 (calculated 510.2724); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.21(s,1H),8.76–7.74(m,2H),8.50(s,1H),7.99(s,2H),7.41–7.31(m,4H),7.24(d,J＝8.6Hz,1H),6.97(dt,J＝15.2,5.9Hz,1H),6.25–6.20(m,2H),3.88(s,3H),3.16(d,J＝5.6Hz,2H),2.65(s,6H),2.31(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.39,154.34,151.35,151.26,146.05,141.39,139.81,137.94,137.19,136.35,136.22,133.92,127.38,126.72,123.51,123.41,122.41,120.69,110.86,109.79,105.12,92.72,60.25,45.43,45.28,33.74；IR ν _max (KBr)cm ^-1 :3456,2943,2853,1675,1585,1512,1366,1236,926,862,772,739。

example 29

1. The same procedures used in example 25, step 2, were repeated except for replacing 4-morpholinyl-3-nitroaniline in example 25, step 2 with an equivalent amount of 4-dimethylamino-3-nitroaniline to give 2- (3-nitro-4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a as a red-orange solid][1,3,5]Triazine 265mg, yield 21.9%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.75(s,1H),8.74(s,1H),8.16(s,1H),8.00(d,J＝0.8Hz,1H),7.81(d,J＝3.2Hz,1H),7.64–7.51(m,2H),7.37–7.30(m,2H),6.95(d,J＝8.7Hz,1H),6.19(s,1H),2.83(s,6H)。

2. with an equivalent mass of 2- (3-nitro-4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 20][1,3,5]Triazine, the other steps were the same as in step 3 of example 20, to give 2- (3-amino-4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 75mg, yield 45.6%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.81(s,1H),8.93(d,J＝8.4Hz,1H),8.05(d,J＝2.0Hz,1H),7.94(d,J＝7.9Hz,1H),7.49(d,J＝7.6Hz,1H),7.45(dd,J＝11.0,3.9Hz,1H),7.28(d,J＝1.4Hz,1H),7.18(s,1H),7.02(d,J＝8.4Hz,1H),7.00–6.89(m,1H),6.26(d,J＝2.0Hz,1H),4.11(s,2H),2.68(s,6H)。

3. with an equivalent amount of 2- (3-amino-4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 28][1,3,5]Triazine, the other steps being the same as in step 3 of example 28, gave compound 29 as a yellow-green solid: 2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 31mg, yield 33.7%, structural characterization data: m.p.168.8-170.4 ℃; HRMS (C) ₂₇ H ₂₈ N ₈ OS)m/z[M+H] ⁺ 513.2180 (calculated 513.2180); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.87(s,1H),9.00(d,J＝7.8Hz,1H),8.76(s,1H),8.52(s,1H),8.06(s,1H),8.00(s,1H),7.93(d,J＝7.8Hz,1H),7.54(d,J＝7.4Hz,1H),7.47(s,2H),7.24(s,1H),6.97(d,J＝15.3Hz,1H),6.29(s,1H),6.21(d,J＝15.3Hz,1H),3.16(d,J＝4.4Hz,2H),2.66(s,6H),2.31(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.49,153.62,151.58,150.02,147.01,141.67,140.86,139.45,138.32,137.31,135.85,133.98,126.56,125.90,125.44,125.29,124.51,122.49,120.77,115.10,110.94,93.36,60.27,45.47,45.24,29.7；IR ν _max (KBr)cm ^-1 :3585,3110,2938,1642,1512,1366,1150,908,778。

example 30

1. This procedure is the same as in example 28, step 1.

2. This procedure is the same as in step 2 of example 28.

3. With equal mass of 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine As an alternative to 2- (4- (4-Methylpiperazino) -3-Aminophenylamino) -4- (1-Methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give a yellow solid compound 30:2- (3-Acrylamido-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 50mg, yield 48.1%, structural characterization data: m.p.199.8-200.3 deg.C; HRMS (C) ₂₅ H ₂₄ N ₈ O)m/z[M+H] ⁺ 453.2141 (calculated 453.2164); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.26(s,1H),8.83–8.77(m,2H),8.51(s,1H),8.02–8.01(m,2H),7.44–7.38(m,3H),7.27(s,1H),7.26(d,J＝2.4Hz,1H),6.44(d,J＝16.9Hz,1H),6.35(dd,J＝16.9,10.5Hz,1H),6.24(d,J＝2.0Hz,1H),5.78(d,J＝10.5Hz,1H),3.94(s,3H),2.67(s,6H)； ¹³ C NMR(151MHz,CDCl3)δ(ppm):163.46,154.29,151.23,145.99,139.81,137.96,137.12,136.39,133.71,131.93,127.33,127.16,123.47,123.34,122.33,122.31,120.65,115.27,110.94,109.75,105.05,92.70,45.26,33.68；IR ν _max (KBr)cm ^-1 :3301,3041,2938,1669,1607,1521,1357,933,864,761,735。

example 31

1. Weighing 1.6g (10 mmol) of 4-fluoro-3-nitroaniline, 1.5g of activated carbon and FeCl ₃ ·6H ₂ 0.1g of O in a round-bottom flask, absolute ethanol was added25mL, heating to 70 ℃, then slowly adding 2.9mL of 80% hydrazine hydrate dropwise into the reaction system, and reacting for 1h at 70 ℃. Filtering, collecting filtrate, drying and concentrating to obtain 1.2g of yellow green solid 4-fluoro-3-amino aniline, wherein the yield is 95.0%, and the structural characterization data is as follows: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):6.74(t,J＝9.7Hz,1H),6.22–5.70(m,2H),3.67(s,2H),3.40(s,2H)。

2. 599mg (2.0 mmol) of 1-methylindole-3-formyl-pyrazol-5-ylthiourea, 376mg (3.0 mmol) of 4-fluoro-3-aminoaniline, 815mg (2.0 mmol) of mercuric chloride and 607mg (6.0 mmol) of triethylamine are weighed into a round-bottomed flask, 15ml of DMF is added, and the mixture is reacted at 100 ℃ for 12 hours in a nitrogen atmosphere. After the reaction is finished, the reaction solution is cooled to room temperature, insoluble substances are removed by suction filtration, and filtrate is collected and refluxed for 12 hours at the temperature of 150 ℃. After the reaction, the reaction mixture was cooled to room temperature, diluted with ethyl acetate and washed with saturated aqueous sodium chloride solution 3 times, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was subjected to silica gel column chromatography (dichloromethane) to give 2- (3-amino-4-fluoroanilino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 150mg, yield 20.1%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.29(s,1H),8.75(d,J＝7.1Hz,1H),8.03(s,1H),7.44–7.43(m,2H),7.39–7.35(m,2H),7.07(s,1H),6.98(t,J＝9.6Hz,1H),6.86(s,1H),6.21(s,1H),3.95(s,3H),3.81(s,2H)。

3. with an equivalent amount of 2- (3-amino-4-fluoroanilino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine in place of 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as in example 28 step 3][1,3,5]Triazine, the other steps being the same as in step 3 of example 28, to give a tan solid compound 31:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoroanilino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 45mg, yield 51.3%, structural characterization data: m.p.202.6-204.3 deg.C; HRMS (C) ₂₇ H ₂₅ FN ₈ O)m/z[M+H] ⁺ 485.2204 (calculated 485.2208); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.24(s,1H),8.74(d,J＝4.8Hz,1H),8.48(s,1H),8.01(s,1H),7.93(s,1H),7.55(s,1H),7.40–7.36(m,3H),7.29(s,1H),7.11(t,J＝9.8Hz,1H),7.03–6.98(m,1H),6.21(s,1H),6.18(d,J＝15.3Hz,1H),3.90(s,3H),3.14(d,J＝5.7Hz,2H),2.30(s,6H)； ¹³ C NMR(151MHz,DMSO-d6)δ(ppm):163.95,154.55,151.05(d, ³ J _C-F ＝4Hz),150.67(d, ¹ J _C-F ＝242Hz),146.72,142.21,140.66,137.47,136.72,127.20,126.31,126.23(d, ³ J _C-F ＝13Hz),123.94,123.67,122.56,116.81,116.21,115.60(d, ² J _C-F ＝20Hz),111.14,104.35,100.00,92.53,60.19,45.57,33.95；IR ν _max (KBr)cm ^-1 :3439,2947,1670,1560,1357,1150,968,847,770。

example 32

1. This step is the same as in step 1 of example 31.

2. The 1-methylindole-3-formyl-pyrazol-5-ylthiourea in step 2 of example 31 was replaced with an equivalent amount of benzothien-3-formyl-pyrazol-5-ylthiourea and the other steps were the same as in step 2 of example 31 to obtain 2- (3-amino-4-fluoroanilino) -4- (benzothien-3-yl) pyrazolo [1,5-a as an orange-red solid][1,3,5]Triazine 127mg, yield 16.7%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.84(s,1H),8.94(d,J＝8.1Hz,1H),8.08(s,1H),7.96(d,J＝7.9Hz,1H),7.53(t,J＝7.5Hz,1H),7.48(t,J＝7.5Hz,1H),7.43(d,J＝7.9Hz,1H),7.12(s,1H),7.06–6.92(m,1H),6.89(s,1H),6.29(s,1H),3.83(s,2H)。

3. with an equivalent mass of 2- (3-amino-4-fluoroanilino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 28][1,3,5]Triazine, the other steps being the same as in step 3 of example 28, to give a tan solid compound 32:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoroanilino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 23mg, yield 26.1%, structural characterization data: m.p.205.5-207.4 ℃; HRMS (C) ₂₅ H ₂₅ FN ₇ OS)m/z[M+H] ⁺ 488.1659 (calculated 488.1663); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.81(s,1H),8.88(s,1H),8.48(s,1H),8.01(s,1H),7.93–7.80(m,2H),7.68–7.64(m,2H),7.44–7.40(m,2H),7.05–6.98(m,2H),6.22(s,1H),6.16(d,J＝13.7Hz,1H),3.09(br s,2H),2.26(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.56,153.52,151.38,149.99,149.29(d, ¹ J _C-F ＝240Hz),147.03,143.36,140.97,139.40,137.24,135.30(d, ⁴ J _C-F ＝2Hz),126.56(d, ³ J _C-F ＝11Hz),125.81,125.36(d, ³ J _C-F ＝15Hz),124.42,122.47,115.55,115.51,114.92(d, ² J _C-F ＝20Hz),113.04,113.02,93.39,60.26,45.49；IR ν _max (KBr)cm ^-1 :3447,3081,2918,1642,1537,1431,1252,1219,991,878,739。

example 33

1. In a round-bottomed flask, 931mg (5.0 mmol) of 4-fluoro-2-methoxy-5-nitroaniline, 766mg (7.5 mmol) of N, N, N' -trimethylethylenediamine and 969mg (7.5 mmol) of N, N-Diisopropylethylamine (DIEA) were weighed, and 20mL of acetonitrile was added to react at reflux temperature for 12 hours. The solvent was evaporated to dryness and the residue was chromatographed on a silica gel column (methanol: dichloromethane =1, 15,v/V) to give 4- (N, N' -trimethylethylenediamino) -2-methoxy-5-nitroaniline 985mg as a tan solid in 73.4% yield with structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):7.24(s,1H),6.59(s,1H),3.89(s,3H),3.76(s,2H),3.18–3.08(m,2H),2.79(s,3H),2.51–2.43(m,2H),2.21(s,6H)。

2. 755mg (2.5 mmol) of 1-methylindole-3-formyl-pyrazol-5-ylthiourea, 150mg (6.2 mmol) of sodium hydride, 677mg (2.5 mmol) of 4- (N, N, N' -trimethylethylenediamine) -2-methoxy-3-nitroaniline and 685mg (2.5 mmol) of mercuric chloride are weighed into a round-bottomed flask, 10mL of DMF is added, and the reaction is carried out at 60 ℃ for 24 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, insoluble matter was removed by suction filtration, and 420mg (7) of calcium oxide was added to the filtrate.5 mmol) were heated at 150 ℃ under reflux for 12h. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with ethyl acetate and washed with a saturated aqueous sodium chloride solution 3 times, the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and subjected to silica gel column chromatography (methanol: dichloromethane =1, 40, V/V) to give 2- (2-methoxy-4- (N, N' -trimethylethylenediamino) -5-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a) as a orange-red solid][1,3,5]Triazine 439mg, yield 34.3%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.27(s,1H),9.23(s,1H),8.77–8.73(m,1H),8.03(d,J＝2.1Hz,1H),7.51(s,1H),7.45–7.33(m,3H),6.68(s,1H),6.27(d,J＝2.1Hz,1H),4.01(s,3H),3.93(s,3H),3.36–3.25(m,2H),2.89(s,3H),2.61–2.53(m,2H),2.28(s,6H)。

3. weighing 2- (2-methoxy-4- (N, N, N' -trimethylethylenediamine) -5-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]439mg (0.85 mmol) of triazine, 94mg of activated carbon and FeCl ₃ ·6H ₂ Adding 25mL of absolute ethyl alcohol into a round-bottom flask by using O5 mg, heating to 70 ℃, then slowly dropwise adding 0.5mL of 80% hydrazine hydrate into the reaction system, and reacting for 1h. The filtrate was suction-filtered, collected, and the organic phase was dried, concentrated and subjected to silica gel column chromatography (methanol: dichloromethane =1, 20, V/V) to give 2- (2-methoxy-4- (N, N' -trimethylethylenediamine) -5-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 298mg, yield 72.1%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.29(s,1H),8.84(dd,J＝6.3,2.5Hz,1H),8.16(s,1H),8.02(d,J＝2.0 Hz,1H),7.66(s,1H),7.51–7.33(m,3H),6.72(s,1H),6.22(d,J＝2.0Hz,1H),3.96(s,3H),3.89(s,3H),3.13(t,J＝6.3Hz,2H),2.71(s,3H),2.67(br s,2H),2.49(s,6H)。

4. with equal mass of 2- (2-methoxy-4- (N, N, N' -trimethylethylenediamine) -5-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give compound 33 as a yellow solid: 2- (3-Acryloylamino-4- (N, N, N' -tris)Methylethylenediamino) -6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 53mg, yield 42.7%, structural characterization data: m.p.251.7-252.9 ℃; HRMS (C) ₂₉ H ₃₃ N ₉ O ₂ )m/z[M+H] ⁺ 540.2820 (calculated 540.2830); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):10.10(s,1H),9.68(s,1H),9.29(s,1H),8.80(dd,J＝5.7,3.0 Hz,1H),8.02(d,J＝2.1Hz,1H),7.61(s,1H),7.42(dd,J＝5.9,3.0Hz,1H),7.38–7.32(m,2H),6.82(s,1H),6.42(d,J＝16.1Hz,1H),6.35(d,J＝1.5Hz,1H),6.31(dd,J＝17.0,10.2Hz,1H),5.68(dd,J＝10.2,1.2Hz,1H),3.94(s,3H),3.92(s,3H),2.93–2.85(m,2H),2.72(s,3H),2.33–2.29(m,2H),2.27(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):163.28,154.20,151.41,151.36,146.10,145.34,139.81,137.25,136.47,132.49,129.54,127.41,126.12,125.77,123.43,123.26,122.20,112.34,109.77,105.34,104.64,93.23,57.49,56.63,56.13,45.58,43.46,33.73；IR ν _max (KBr)cm ^-1 :3431,3055,2930,1685,1607,1538,1374,1150,770,545。

example 34

1. In a round-bottomed flask, 1.0g (5.4 mmol) of 4-fluoro-2-methoxy-5-nitroaniline, 818. Mu.L (6.0 mmol) of a tetrahydrofuran solution of dimethylamine and 1.0mL (6.0 mmol) of N, N-Diisopropylethylamine (DIEA) were weighed, and 25mL of tetrahydrofuran was added to react at 50 ℃ for 12 hours. The solvent was evaporated under reduced pressure and separated by silica gel column chromatography (methanol: dichloromethane =1, 20,v/V) to give 4-dimethylamino-2-methoxy-5-nitroaniline 793mg as a dark brown oil in 69.5% yield with the structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):7.35(d,J＝3.9Hz,1H),6.46(d,J＝2.8Hz,1H),3.92(s,3H),3.71(s,2H),2.83(s,6H)。

2. 830mg (2.77 mmol) of 1-methylindole-3-formyl-pyrazol-5-ylthiourea, 650mg (3.0 mmol) of 4-dimethylamino-2-methoxy-5-nitroaniline, 816mg (3.0 mmol) of mercuric chloride and 607mg (6.0 mmol) of triethylamine are weighed in a round-bottomed flask, and added15ml of DMMF, and reacting for 12 hours at 100 ℃ in a nitrogen atmosphere. After completion of the reaction, the reaction mixture was cooled to room temperature, insoluble matter was removed by suction filtration, and 453mg (8.1 mmol) of calcium oxide was added to the filtrate, and the mixture was heated under reflux at 150 ℃ for 10 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with ethyl acetate and washed with a saturated aqueous sodium chloride solution 3 times, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to silica gel column chromatography (methanol: dichloromethane =1, 100, V/V) to give 2- (4-dimethylamino-2-methoxy-5-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as an orange-red solid][1,3,5]Triazine 391mg, yield 30.8%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.32(s,1H),9.28(s,1H),8.82–8.75(m,1H),8.05(d,J＝2.0Hz,1H),7.50(s,1H),7.47–7.43(m,1H),7.43–7.34(m,2H),6.52(s,1H),6.29(d,J＝2.0Hz,1H),4.04(s,3H),3.97(s,3H),2.94(s,6H)。

3. with equal mass of 2- (4-dimethylamino-2-methoxy-5-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (2-methoxy-4- (N, N, N' -trimethylethylenediamine) -5-nitrophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a ] in step 3, example 33][1,3,5]Triazine, the other steps were the same as in step 3 of example 33, to give 2- (4-dimethylamino-2-methoxy-5-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a yellow-green solid][1,3,5]Triazine 309mg, yield 84.9%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.26(s,1H),8.83(d,J＝6.8Hz,1H),8.13(s,1H),8.01(d,J＝1.6Hz,1H),7.61(s,1H),7.45–7.33(m,3H),6.71(s,1H),6.20(s,1H),3.93(s,3H),3.90(s,3H),3.88(s,2H),2.69(s,6H)。

4. with an equal amount of 2- (4-dimethylamino-2-methoxy-5-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (4- (4-methylpiperazino) -3-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 4 of example 20][1,3,5]Triazine, the other steps being the same as in step 4 of example 20, to give compound 34 as a yellow solid: 2- (3-Acrylamido-4-dimethylamino-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 65mg, yield 58.4%, structural characterization data: m is.p.213.4–214.8℃；HRMS(C ₂₆ H ₂₆ N ₈ O ₂ )m/z[M+H] ⁺ 483.2243 (calculated 483.2251); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.58(s,1H),9.25(s,1H),8.82–8.72(m,1H),8.48(s,1H),8.01(d,J＝2.0Hz,1H),7.55(s,1H),7.45–7.37(m,1H),7.37–7.28(m,2H),6.78(s,1H),6.46–6.27(m,3H),5.72(d,J＝10.8Hz,1H),3.92(s,3H),3.90(s,3H),2.66(s,6H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):162.93,154.23,151.39,151.34,146.07,145.55,139.84,137.90,137.22,132.26,127.37,126.69,126.34,125.43,123.39,123.26,122.19,112.55,109.79,105.26,102.77,93.12,56.12,45.03,33.71；IR ν _max (KBr)cm ^-1 :3430,3068,2938,1676,1607,1547,1374,1158,770,545。

example 35

1. The same procedures as in step 1 of example 31 were repeated except for replacing 4-fluoro-3-nitroaniline in step 1 of example 31 with an equivalent amount of 4-fluoro-2-methoxy-5-nitroaniline to give 1.4g of a crude yellowish green solid, 4-fluoro-2-methoxy-5-aminoaniline, in a crude yield of 89.7%, and having the structural characterization data as follows: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):6.48(dd,J＝11.9,3.3Hz,1H),6.12(dd,J＝8.9,3.3Hz,1H),3.68(s,3H),3.37(s,3H)。

2. the same procedures used in example 33, step 2, were repeated except for replacing 4- (N, N, N' -trimethylethylenediamine) -2-methoxy-3-nitroaniline in example 33, step 2, with an equivalent amount of 4-fluoro-2-methoxy-5-aminoaniline to give 2- (4-fluoro-2-methoxy-5-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a as a beige solid][1,3,5]Triazine 85mg, yield 8.4%, structural characterization data: ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.30(s,1H),8.83(dd,J＝5.9,3.0Hz,1H),8.28(d,J＝9.6Hz,1H),8.04(d,J＝2.0Hz,1H),7.60(s,1H),7.49–7.36(m,3H),6.69(d,J＝11.8Hz,1H),6.23(d,J＝2.0Hz,1H),3.97(s,3H),3.89(s,3H),3.56(s,2H)。

3. with equal mass of 2- (4-fluoro-2-methoxy-5-aminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine for substitution of 2- (3-amino-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a in step 3 of example 28][1,3,5]Triazine, the other steps are the same as in example 28, step 3. The obtained crude product is separated by column chromatography to obtain a tan solid compound 35:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoro-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 32mg (yield 34.8%) and compound 36:2- (3- (N-methylpyrrolidin-2-yl) imino-4-fluoro-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine 7mg (yield 8.0%). Structural characterization data for compound 35 is: m.p.169.7-170.8 ℃; HRMS (C) ₂₇ H ₂₇ FN ₈ O ₂ )m/z[M+H] ⁺ 515.2303 (calculated 515.2314); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.32(br s,1H),9.29(s,1H),8.82–8.74(m,1H),8.03(d,J＝2.0Hz,1H),7.53(s,1H),7.46–7.41(m,1H),7.41–7.34(m,2H),7.23(s,1H),7.04–6.93(m,1H),6.73(d,J＝11.7Hz,1H),6.29(s,1H),6.16(d,J＝15.3Hz,1H),3.95(s,3H),3.93(s,3H),3.12(d,J＝4.5Hz,2H),2.29(s,6H)； ¹³ C NMR(151MHz,DMSO-d6)δ(ppm):163.83,155.65,153.19(d, ¹ J _C-F ＝244Hz),151.49,151.09,150.74,150.69,146.82,141.90,140.65,137.45,127.14,125.95,123.99(d, ⁴ J _C-F ＝3Hz),123.62,122.51,122.13,118.13(d, ³ J _C-F ＝13Hz),111.21,104.33,100.78(d, ² J _C-F ＝25Hz),91.92,60.26,56.76,45.62,33.97；IR ν _max (KBr)cm ^-1 3422,3059,2938,1676,1616,1538,1374,1158,769,553; the structural characterization data for compound 36 is: m.p.104.3-105.7 ℃; HRMS (C) ₂₇ H ₂₇ FN ₈ O ₂ )m/z[M+H] ⁺ 485.2204 (calculated 485.2214); ¹ H NMR(600MHz,CDCl ₃ )δ(ppm):9.30(s,1H),8.83(d,J＝8.1Hz,1H),8.28(d,J＝9.2Hz,1H),8.03(s,1H),7.61(s,1H),7.49–7.37(m,3H),6.72(d,J＝11.6Hz,1H),6.22(s,1H),3.97(s,3H),3.94(s,3H),3.43(t,J＝6.9Hz,2H),3.07(s,3H),2.53(t,J＝7.8Hz,2H),2.02–1.93(m,2H)； ¹³ C NMR(151MHz,CDCl ₃ )δ(ppm):164.68,154.17,151.31,150.78(d, ¹ J _C-F ＝238Hz),146.02,144.28,144.27,144.23,139.80,137.20,127.37,124.62,124.60(d, ⁴ J _C-F ＝2Hz),123.29,122.38,116.29,109.82,105.18,99.36(d, ² J _C-F ＝27Hz),92.56,56.29,51.85,33.74,31.71,28.34,19.68；IR ν _max (KBr)cm ^-1 :3062,2952,1617,1519,1423,1372,894,767。

example 36

The invention discloses an application of pyrazolo [1,5-a ] [1,3,5] triazine derivatives and salts thereof in preparing anticancer drugs, and the specific test conditions are as follows:

1. cell line

Human colon cancer cells SW480, human non-small cell lung cancer cells A549, human skin squamous cancer cells A431 and human lung cancer cells NCI-H1975 were all purchased from Shanghai cell Bank of Chinese academy of sciences.

2. Reagents and materials

MTT (MPBIO), 96-well cell culture plates, fetal bovine serum (Gibco), DMEM (Dulbecco's Modified Eagle Medium powder, high glucose, gibco BRL, gibco), penicillin, streptomycin (bi yun), trypsin digest (bi yun), and microplate reader (penespire).

3. Experimental procedure

(1) Cell culture

The cell culture medium used for the cancer cells SW480, A549, A431 and NCI-H1975 in the experiments was DMEM medium containing 10% (v/v) fetal bovine serum, 100units/mL penicillin, 100. Mu.g/mL streptomycin and 2 mmol/LL-glutamine. The cells were subjected to saturation humidity at 37 ℃ and 5% CO ₂ Culturing in an incubator. Passage is carried out once every 2 to 3 days.

(2) Detection of anti-cancer Activity

The growth inhibitory activity of compounds 1 to 36 on cancer cells was measured by the MTT method. Respectively taking human cancer cells in logarithmic growth phase, digesting with 0.25% trypsin digestive juice, centrifuging, resuspending, counting,preparing cell suspension, adjusting the concentration of the cell suspension to 2.0 × 10 ⁴ ～5×10 ⁴ one/mL. Inoculating the cell suspension into 96-well culture plates (100. Mu.L/well), saturating at 37 ℃ and 5% CO ₂ Culturing in an incubator for 24h. Test compounds were diluted with cell culture medium to the desired concentration, added to 96-well plates (100. Mu.L/well) inoculated with human cancer cells at a final DMSO concentration of 0.5%, and placed in an incubator for 72h. MTT was added to 96-well plates (20. Mu.L/well) and reacted in an incubator for 4h. The liquid in the wells was removed by blotting, DMSO (150 μ L/well) was added, and the formazan was shaken on a shaker for 10min to dissolve the formazan completely. Then, the absorbance (OD value) at the wavelength of 570nm was measured by a microplate reader, the absorbance at the wavelength of 630nm was used as a reference, and the cell growth inhibition rate was calculated using the corresponding solvent as a control.

The inhibition rate of the test compounds on the growth of cancer cells was calculated as follows:

cancer cell growth inhibition rate% = [1- (ODs-OD) _NC )/(OD _PC -OD _NC )]×100％

Wherein: OD _S The absorbance values (cells + test compound + MTT) for the sample wells are expressed; OD _PC The absorbance values (cells + DMSO + MTT) for the control wells are expressed; OD _NC Represent absorbance values for the zeroed wells (cell culture medium + DMSO + MTT); ODs = OD _570s -OD _630s ；OD _PC ＝OD _570PC -OD _630PC ；OD _NC ＝OD _570NC -OD _630NC 。

Fitting of test Compounds to cancer cell growth inhibition curves and IC ₅₀ The calculation of (2): fitting the inhibition curve of the tested compound on the growth of the cancer cells by adopting Graphpad Prism5, and obtaining IC ₅₀ The value is obtained. Each set was provided with 3 replicate wells, repeated at least 3 times.

4. Results of the experiment

TABLE 1 Activity of test Compounds to inhibit cancer cell proliferation [ IC ₅₀ (μmol/L)]

The data in table 1 show that the compounds of the present invention have significant antiproliferative activity against at least one of the four cancer cells tested, most of the compounds showed good antiproliferative activity against the four cancer cells tested, and in particular, compounds 24, 28, 31, 32, 33 had prominent proliferation inhibitory activity against the four cancer cells tested. The results show that the pyrazolo [1,5-a ] [1,3,5] triazine derivative and the salt thereof have anticancer activity and can be used for preparing anticancer drugs.

Claims

1. Pyrazolo [1,5-a][1,3,5]Triazine derivatives and salts thereof, characterized in that the derivatives have the following structural formula:

in the formula, R' is any one of benzothiophen-3-yl and 1-methylindol-3-yl; r is-NHCH ₂ CH ₂ R', diethylaminoethoxy,N,N,N' -trimethylethylenediamino, 4- (2-methoxyethyl) piperazinyl, diethylamino, 4-methylpiperazinyl, 4-acryloylpiperazinyl,

Wherein R' is any one of diethylamino, morpholinyl, 4-methylpiperazinyl and pyrrolidinyl, and R ¹ Is acrylamido, 4- (dimethylamino) but-2-enamidoamino,N-any one of methylpyrrolidin-2-ylimino; r ² Is a substituent attached to any position of the benzene ring, R ² Selected from fluoro, methoxy, dimethylamino, morpholinyl, diethylamino, 4-methylpiperazinyl,N,N,N' -trimethylethylene diamino, n is 1 or 2.

2. According to the rightPyrazolo [1,5-a][1,3,5]Triazine derivatives and salts thereof, characterized in that: the salt is pyrazolo [1,5-a][1,3,5]Triazine derivatives are salts with hydrochloric acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, maleic acid, fumaric acid, formic acid, acetic acid and benzoic acid, respectively.

3. Pyrazolo [1,5-a][1,3,5]Triazine derivatives and salts thereof, characterized in that: the salt is pyrazolo [1,5-a][1,3,5]Salts of triazine derivatives with p-toluenesulfonic acid.

4. Pyrazolo [1,5-a][1,3,5]Triazine derivative, characterized in that the derivative is any one of the following compounds 1 to 36:

compound 1:2- (2-diethylaminoethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 2:2- (2-morpholinoethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 3:2- (2- (4-methylpiperazino) ethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 4:2- (4- (2-methoxyethyl) piperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 5:2- (4-methylpiperazino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 6: 2-piperazinyl-4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 7:2- (4-acryloylpiperazinyl) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 8:2- (4-methylpiperazino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine p-methyl benzene sulfonate

Compound 9:2- (2-pyrrolidinoethylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 10:2- (2-diethylaminoethoxy) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 11:2- (2-diethylaminoethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 12:2- (2-Morpholinylethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 13:2- (4- (2-methoxyethyl) piperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 14:2- (4-methylpiperazino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 15: 2-piperazinyl-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 16:2- (4-acryloylpiperazinyl) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 17:2- (2-pyrrolidinoethylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 18: 2-diethylamino-4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 19:2- (N,N,N' -trimethylethylenediamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 20:2- (3-Acrylamido-4- (4-methylpiperazino) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 21:2- (3-Acrylamido-4-diethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 22:2- (3-Acryloylamino-4-morpholinophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 24:2- (3-acrylamido-4-) (N,N,N' -trimethylethylenediamino) phenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 25:2- (3-Acrylamido-4-morpholinophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 26:2- (3-Acrylamido-4- (4-methylpiperazino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 27:2- (3-acrylamido-4-) (N,N,N' -trimethylethylenediamino) phenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 28:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-dimethylaminobenzamido) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 29:2- (3- (4-dimethylamino butane-2-Enoylamino) -4-dimethylaminophenylamino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 30:2- (3-Acrylamido-4-dimethylaminophenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 31:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoroanilino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 32:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoroanilino) -4- (benzothien-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 33:2- (3-acrylamido-4-) (N,N,N' -trimethylethylenediamino) -6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 34:2- (3-Acryloylamino-4-dimethylamino-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 35:2- (3- (4-dimethylaminobutyl-2-enoylamino) -4-fluoro-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

Compound 36:2- (3- (N-methylpyrrolidin-2-yl) imino-4-fluoro-6-methoxyphenylamino) -4- (1-methylindol-3-yl) pyrazolo [1,5-a][1,3,5]Triazine

。

5. Pyrazolo [1,5-a][1,3,5]The use of triazine derivatives and salts thereof in the preparation of anti-cancer drugs.

6. Pyrazolo [1,5-a][1,3,5]The application of triazine derivatives and salts thereof in preparing anticancer drugs is characterized in that: the cancer is any one of colon cancer, lung cancer and skin squamous carcinoma.