CN105906633B - Pyrazoles N phenyl maleimide derivatives containing chromone structure and preparation method and application - Google Patents

Abstract

The invention discloses a 3-(6-bromo-4-oxygen-4H-chromone)-1,5-diphenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4 , 6(3aH,5H)-diketone and its preparation method and application, the preparation method uses 1,3-dipolar cycloaddition method to introduce a pyrazole ring into the N-substituted phenylmaleimide structure, and The chromone derivative reaction introduces the chromone structure, thereby synthesizing a novel 3-(6-bromo-4-oxygen-4H-chromone)-1,5-diphenyl-1,6a-dihydropyrroline[3 ,4-c]pyrazole-4,6(3aH,5H)-dione, the above compound has good inhibitory effect on tumor cell lines, and has better inhibitory rate and selectivity for oral cancer cells. It has very good industrial application prospects in terms of tumor drugs and the like.

Description

Pyrazole N-phenylmaleimide derivatives containing chromone structure and preparation method thereof Law and Application

technical field

The present invention relates to pyrazole N-phenylmaleimide derivatives containing chromone structure, specifically 3-(6-bromo-4-oxygen-4H-chromone)-1,5-diphenyl -1,6a-dihydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione and its preparation method and application.

Background technique

N-phenylmaleimide, chemical name: 1-phenylpyrrolidine-2,5-dione, chemical structure as follows:

1-Phenylpyrrolidine-2,5-dione

The 1,3-dipolar cycloaddition reaction has become the most important method for the synthesis of five-membered heterocyclic compounds due to its good regio and host selectivity, and it is also an active class of reactions in the research of heterocyclic medicinal chemistry. In recent years, due to the wide range of biological activities of chromone, such as anticancer, antibacterial, inhibition of platelet aggregation and so on, it has attracted much attention. Therefore, no matter from the perspective of pharmacology or synthesis, this kind of heterocyclic compounds has high synthetic value.

Pyrazole derivatives have attracted widespread attention as a class of useful intermediates and their various pharmaceutical activities. The present invention provides a brand-new pyrazole N-phenylmaleimide derivative containing a chromone structure by studying the effect of aggregation of different heterocyclic rings in the same molecule on pharmacological activity.

Contents of the invention

The purpose of the first aspect of the present invention is to provide a new pyrazole N-phenylmaleimide derivative containing a chromone structure.

The technical scheme that the present invention takes is as follows:

A pyrazole N-phenylmaleimide derivative containing a chromone structure, its chemical name is: 3-(6-bromo-4-oxygen-4H-chromone)-1,5-diphenyl Base-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione, its structural formula is as follows:

The relevant experimental data of this compound are as follows:

The applicant found through research that: on the basis of introducing the five-membered pyrazole ring into N-phenylmaleimide, the structure of chromone was introduced, and a new pyrazole N-benzene compound containing chromone structure was obtained. Based maleimide derivatives, the compound can alter the pharmacological activity.

The second aspect object of the present invention is to provide a kind of preparation method of the above-mentioned pyrazole N-phenylmaleimide derivative containing chromone structure, it is characterized in that, comprises the following steps: maleic anhydride and Aniline reaction synthesizes N-phenylmaleimide (3), then adopts 3-position to have formaldehyde to replace 6-bromochromone and phenylhydrazine dehydration reaction to synthesize 6-bromochromone phenylhydrazone (4), and finally N-phenylmaleimide and 6-bromochromone phenylhydrazone carry out dipolar cycloaddition reaction to synthesize pyrazole N-phenylmaleimide derivatives containing chromone structure (5), That is, 3-(6-bromo-4-oxo-4H-chromone)-1,5-diphenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6(3aH, 5H)-diketones.

The reaction equation that the present invention relates to is as follows:

Further, the preparation method of a pyrazole N-phenylmaleimide derivative containing a chromone structure is characterized in that it comprises the following steps:

(1), the synthesis of N-phenylmaleimide:

Dissolve maleic anhydride and aniline in acetone solvent, react under stirring, and gradually produce a light yellow precipitate. After reacting at room temperature for 2 hours, add manganese acetate, triethylamine and acetic anhydride in sequence. After heating up, the precipitate gradually dissolves. React at 50-60°C for 2 hours, the solution turns from orange to red-black, and a new precipitate, namely N-phenylmaleimide (3), gradually appears, cool to room temperature, wash the precipitate with water, dry, and reconstitute with acetone Crystallization obtains product N-phenylmaleimide (3);

(2), the synthesis of 6-bromochromone phenylhydrazone:

The 6-bromochromone phenylhydrazone (4) is a known compound, and its CAS number is 68287-82-1. The present invention adopts the dehydration reaction of 6-bromochromone and phenylhydrazine to form a seat Synthesized by the method of Huofu base: take 2mmol of phenylhydrazine and add it into a flask containing 10mL of tetrahydrofuran, heat and reflux and stir until dissolved, then slowly add 20mL of anhydrous Ethanol solution, continue to reflux and stir for 1 hour, add 10 drops of hydrochloric acid dropwise, a light yellow precipitate appears, continue to heat and reflux and stir for 5 hours, stop heating, add 20 mL of distilled water after cooling to room temperature and stir, the light yellow precipitate becomes darker, suction filter to obtain 6 -Bromochromone phenylhydrazone (4), a yellow-red needle-like product; washed with anhydrous ether several times, and vacuum-dried to obtain the product 6-bromochromone phenylhydrazone (4);

(3), import chromone structure:

Dissolve N-phenylmaleimide and 6-bromochromone phenylhydrazone in absolute ethanol, add chloramine T, reflux for 12 hours, carry out 1,3-dipolar cycloaddition reaction, import Chromone and pyrazole structures, recrystallized with methanol, and dried in vacuo to obtain compound (5).

Further settings are:

In the step (1), the pale yellow precipitate obtained from the reaction needs to be filtered under reduced pressure.

The object of the third aspect of the present invention is to provide the application of the aforementioned pyrazole N-phenylmaleimide derivatives containing chromone structure in the preparation of antitumor drugs. Experimental verification: the above compounds have inhibitory effects on different tumor lines such as human liver cancer cells, oral cancer cells, gastric cancer cells, ovarian cancer cells, leukemia cells, intestinal cancer cells, etc., among which KB (oral cancer cells) have more inhibitory effects. With excellent inhibition rate and selectivity, it can be used to prepare anti-tumor drugs alone, and can also be used as an active ingredient to prepare anti-tumor compositions with other anti-tumor drugs, and has very good industrial application prospects.

The beneficial effects of the present invention are as follows:

The applicant has discovered through research that introducing the structure of chromone on the basis of N-phenylmaleimide introducing the five-membered pyrazole ring can change the pharmacological activity. In view of the effect of aggregation in molecules on pharmacological activity, the applicant synthesized pyrazole N-phenylmaleimide derivatives containing chromone structure through dipolar cycloaddition reaction. The keto-structure pyrazole N-phenylmaleimide derivative has very good industrial application prospects in the preparation of antitumor drugs.

detailed description

Chloramine T: English name Chloramine-T, chemical structure is

The present invention will be further described below in conjunction with the examples, but the examples should not be construed as limiting the scope of the present invention.

Embodiment 1: the synthesis of N-phenylmaleimide:

Dissolve maleic anhydride and aniline in a certain amount of acetone solvent, add the aniline solution dropwise to the reaction flask containing the maleic anhydride solution under stirring, the reaction is exothermic and gradually produces a light yellow precipitate, and react at room temperature for 2 After one hour, manganese acetate, triethylamine and acetic anhydride were added in sequence. After the temperature was raised, the precipitate gradually dissolved and reacted at 50-60°C for 2 hours. , cooled to room temperature, the precipitate in water was washed with a large amount of water, dried, and recrystallized with acetone to obtain the product N-phenylmaleimide (3);

Embodiment 2: the synthesis of 6-bromochromone phenylhydrazone:

The method of dehydration reaction of 6-bromochromone with formaldehyde group in the 3 position and phenylhydrazine to generate Schiff base: take 2mmol of phenylhydrazine and add it to a flask containing 10mL of tetrahydrofuran, heat and reflux and stir until dissolved, then slowly add 20mL of Dissolve 2 mmol of anhydrous ethanol solution with formaldehyde group at position 3 of 6-bromochromone, continue to reflux and stir for 1 hour, add 10 drops of hydrochloric acid dropwise, and a pale yellow precipitate appears. Continue to heat and reflux and stir for 5 hours, stop heating, add 20 mL of distilled water after cooling to room temperature and stir, the pale yellow precipitate becomes darker, and suction-filters to obtain 6-bromochromone phenylhydrazone, a yellow-red needle-like product. Rinse with anhydrous ether several times, and vacuum dry to obtain the product 6-bromochromone phenylhydrazone (4);

Example 3: 3-(6-bromo-4-oxo-4H-chromone)-1,5-diphenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6 Synthesis of (3aH,5H)-diketones:

Add 1mmol N-phenylmaleimide and 1.1mmol 6-bromochromone phenylhydrazone to 20mL ethanol, then add 1.2mL chloramine T, and reflux for 12 hours. The precipitate was filtered, washed with 95% ethanol, recrystallized with methanol, and dried in vacuum to obtain the product. The product is a yellow crystal with a yield of 44.3%. The product has been detected: the molecular formula is C ₂₆ H ₁₆ BrN ₃ O ₄ , and the name is: 3-(6-bromo-4-oxo-4H-chromone)-1,5-di Phenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione (Compound 5), mp 139.5-141°C.

Confirmed structure: C ₂₆ H ₁₆ BrN ₃ O ₄

¹ H NMR(DMSO)δ:7.288-7.492(m,13H,Ar-H),6.47(s,1H,C=CH),5.82(d,J=1010Hz,1H),5.34(d,J=1014Hz ,1H)

IR 3457(NC＝O), 3085(ArH), 1720(C＝O), 1575(C＝N), 1293(COC)cm ^-1

m/e: 513.03 (100.0%), 515.03 (97.6%).

_{Anal.calcd.for C26H16BrN3O4} : C, 60.72 _; H, _3.16 ; N, 8.15.

Application Example 4: MTT method was used to detect the antitumor activity of the test compound on different tumor lines.

With the compound (5) prepared in the above-mentioned examples, different tumor strains (tumor cells Bel-7402, KB, SGC7901, HO8901, HL-60, ECA109) were used as experimental objects respectively, and the in vitro effects of the test compound (5) on different tumor strains were tested. Inhibition: The experiment adopts tetramethylazolate microenzyme reaction colorimetry (MTT method), and the activity is represented by the half inhibitory concentration (IC ₅₀ ).

The specific experimental steps are as follows:

Compound 5 was dissolved in DMSO, diluted, tumor cells Bel-7402 (human liver cancer cells), KB (oral cancer cells), SGC7901 (stomach cancer cells), HO8901 (ovarian cancer cells), HL-60 (leukemia cells), ECA109 ( Intestinal cancer cells) Seed 4000 cells/200 μL/well on a 96-well plate, add 2 μL of compound to each well, the final concentration is 12.0 μM, 6.0 μM, 3.0 μM, 1.5 μM, and culture the cells together at 37°C, 5% CO ₂ Incubate in the box for 72 hours, and use DMSO (1%) as a blank control. After 72 hours, add MTT with a final concentration of 0.25mg/mL, place in a 37°C, 5% _CO2 cell culture incubator for 4 hours, then blot the solvent, add 100μl DMSO to each well, and use an enzyme-linked immunosorbent analyzer at 570nm The absorbance (OD value) was measured, and the obtained data was used to calculate the IC ₅₀ value.

The test compounds with different concentrations were screened with a 96-well plate, and the IC ₅₀ value was calculated according to the obtained inhibition rate, and the results are shown in the table below.

Table 1. IC ₅₀ values of pyrazole N-phenylmaleimide derivatives containing chromone structure against six tumor cell lines.

It can be seen from the data in the above table that the compounds prepared by the present invention have inhibitory effects on six tumor cell lines, among them, they have better inhibitory rate and selectivity for KB (oral cancer cells), and can independently prepare antitumor drugs, It can also be used as an active ingredient and other antitumor drugs to prepare antitumor compositions, and has very good industrial application prospects.

Claims (5)

1. the pyrazole N-phenylmaleimide derivative that contains chromone structure, its structural formula is as follows: 2. a kind of preparation method that contains the pyrazole N-phenylmaleimide derivative of chromone structure, is characterized in that, comprises the following steps: maleic anhydride and aniline are reacted synthetic N-phenylmaleimide to imide, and then use 3-formaldehyde substituted 6-bromochromone and phenylhydrazine dehydration reaction to synthesize 6-bromochromone phenylhydrazone, and finally N-phenylmaleimide and 6-bromo Chromone phenylhydrazone undergoes dipolar cycloaddition reaction to synthesize pyrazole N-phenylmaleimide derivatives containing chromone structure. 3. the preparation method of a kind of pyrazole N-phenylmaleimide derivative containing chromone structure according to claim 2, is characterized in that, comprises the following steps: (1) Synthesis of N-phenylmaleimide: Dissolve maleic anhydride and aniline in acetone solvent, react under stirring, and gradually produce a light yellow precipitate. After reacting at room temperature for 2 hours, add manganese acetate, triethylamine and acetic anhydride in sequence. After heating up, the precipitate gradually dissolves. React at 50-60°C for 2 hours, the solution turns from orange to red-black, and a new precipitate, N-phenylmaleimide, gradually appears, cool to room temperature, wash the precipitate with water, dry, and recrystallize with acetone to obtain the product N-phenylmaleimide; (2) Synthesis of 6-bromochromone phenylhydrazone: Take 2mmol of phenylhydrazine and add it to a flask containing 10mL of tetrahydrofuran, heat and reflux and stir until dissolved, then slowly add 20mL of anhydrous ethanol solution in which 2mmol of 6-bromochromone substituted with formaldehyde group at the 3-position is added dropwise, and continue to reflux and stir 1h, add 10 drops of hydrochloric acid dropwise, a pale yellow precipitate appears, continue to heat and reflux and stir for 5h, stop heating, add 20mL distilled water after cooling to room temperature and stir, the pale yellow precipitate becomes darker, and 6-bromochromone benzene is obtained by suction filtration Hydrazone, a yellow-red needle-like product; washed with anhydrous ether several times, and vacuum-dried to obtain the product 6-bromochromone phenylhydrazone; (3) Import the chromone structure: Dissolve N-phenylmaleimide and 6-bromochromone phenylhydrazone in absolute ethanol, add chloramine T, reflux for 12 hours, carry out 1,3-dipolar cycloaddition reaction, import Chromone and pyrazole structures, recrystallized with methanol, and vacuum-dried to obtain pyrazole N-phenylmaleimide derivatives containing chromone structures. 4. the preparation method of a kind of pyrazole N-phenylmaleimide derivative containing chromone structure according to claim 3 is characterized in that: in the described step (1), the reaction obtained The pale yellow precipitate needs to be filtered under reduced pressure. 5. The application of the compound according to claim 1 in the preparation of antitumor drugs.