CN105859724B - Rubigan 1,6a pyrrolin quinoline [3,4 c] pyrazoles 4,6 (3aH, 5H) diketone of 3 (the oxygen 4H chromones of 6 bromine 4) 1 phenyl 5 and preparation method and application - Google Patents

Abstract

The invention discloses a 3-(6-bromo-4-oxygen-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c] Pyrazole-4,6(3aH,5H)-dione and its preparation method and application, the preparation method uses 1,3-dipolar cycloaddition method to introduce five-membered N-p-chlorophenylmaleimide On the basis of the pyrazole ring, the structure of chromone was introduced to synthesize a new type of 3-(6-bromo-4-oxygen-4H-chromone)-1-phenyl-5-p-chlorophenyl-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, in the preparation of antitumor drugs, etc., has a very good industrial application prospect.

Description

3-(6-Bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyridine Porzoline[3,4-c]pyrazole-4,6(3aH,5H)-dione and its preparation method and application

technical field

The invention relates to a new pyrazole N-p-chlorophenylmaleimide derivative containing a chromone structure, specifically 3-(6-bromo-4-oxygen-4H-chromone)-1 -Phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione and its preparation method and application.

Background technique

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

1-p-Chlorophenylpyrrolidine-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-p-chlorophenylmaleimide 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-p-chlorophenylmaleimide derivative containing a chromone structure.

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

A pyrazole N-p-chlorophenylmaleimide derivative containing a chromone structure, its chemical name is: 3-(6-bromo-4-oxygen-4H-chromone)-1-phenyl -5-p-chlorophenyl-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:

Through research, the applicant found that: using chromone derivatives to substitute on the structure of pyrazole and modifying the structure of N-p-chlorophenylmaleimide, a new pyrazole containing chromone structure was obtained N-p-chlorophenylmaleimide derivative, the compound can change the pharmacological activity.

The second aspect object of the present invention is to provide a kind of preparation method of above-mentioned pyrazole N-p-chlorophenylmaleimide derivative containing chromone structure, it is characterized in that, comprises the following steps: Acid anhydride and p-chloroaniline react to synthesize N-p-chlorophenylmaleimide (3), and then adopt 3-position substituted 6-bromochromone and phenylhydrazine dehydration reaction to synthesize 6-bromochromone benzene Hydrazone (4), finally carry out dipolar cycloaddition reaction with N-p-chlorophenylmaleimide (3) and 6-bromochromone phenylhydrazone (4), synthesize 3-(6-bromo-4 -Oxygen-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione (5).

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

Further, the described 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4- c] the preparation method of pyrazole-4,6(3aH,5H)-dione, is characterized in that, comprises the following steps:

(1), the synthesis of N-p-chlorophenylmaleimide:

Dissolve maleic anhydride and p-chloroaniline in acetone solvent, react under stirring, and gradually produce a light yellow precipitate, react at room temperature for 1 hour, take a small amount of precipitate, wash with water, dry, add manganese acetate, triethyl Amine and acetic anhydride, after heating up, the precipitate gradually dissolves, react at 50-60°C for 2 hours, the solution turns from orange to red and black, and new precipitates gradually appear, cool to room temperature, wash the precipitate with water, dry, and recrystallize with acetone Obtain product N-p-chlorophenylmaleimide (3);

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

The 6-bromochromone phenylhydrazone (4) is a known compound with a CAS number of 68287-82-1. The present invention uses 6-bromochromone substituted with formaldehyde at the 3-position and dehydration reaction with phenylhydrazine to generate Schiff base method for synthesis: take 2mmol of phenylhydrazine and add it to a flask containing 10mL of tetrahydrofuran, stir in a boiling water bath under reflux until dissolved, then slowly add 20mL of 6-bromochromone dissolved in 2mmol of 3-position formaldehyde group substituted For absolute ethanol solution, continue to reflux and stir in a boiling water bath for 1 hour, add 10 drops of hydrochloric acid dropwise, and a light yellow precipitate appears; continue to reflux and stir in a boiling water bath for 5 hours, stop the water bath, add 20 mL of distilled water and stir, the light yellow precipitate becomes darker, and is obtained by suction filtration 6-bromochromone phenylhydrazone, a yellow-red needle-like product, was washed with anhydrous ether several times, and vacuum-dried to obtain the product 6-bromochromone phenylhydrazone (4);

(3), 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c]pyrazole Synthesis of -4,6(3aH,5H)-dione:

Add 1mmol N-p-chlorophenyl maleimide and 1.1mmol chromone phenylhydrazone to 20mL ethanol, then add 1.2mL chloramine T, and reflux for 12 hours. The precipitate was filtered, washed, recrystallized with methanol, and dried in vacuo to obtain the product 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-N-p-chlorophenyl-1,6a-di Hydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione (5).

Further settings are:

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

The third aspect object of the present invention is to provide a kind of 3-(6-bromo-4-oxygen-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline [3 ,4-c]Application of pyrazole-4,6(3aH,5H)-dione 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., and have a better inhibitory rate on leukemia cells And selectivity, it can 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 found through research that a new pyrazole containing chromone structure was obtained by substituting chromone derivatives on the structure of pyrazole and transforming the structure of N-p-chlorophenylmaleimide. N-p-chlorophenylmaleimide derivatives, this compound can change the pharmacological activity. After further experiments and analysis, the influence of different heterocycles on the pharmacological activity caused by the aggregation of different heterocycles in the same molecule was studied. The pyrazole N-p-chlorophenylmaleimide derivatives containing chromone structure were synthesized by polar cycloaddition reaction, and the new pyrazole N-p-chlorophenylmaleimide derivatives containing chromone structure prepared by this method The imide derivatives have 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-p-chlorophenylmaleimide:

Dissolve maleic anhydride and p-chloroaniline in a certain amount of acetone solvent, add the p-chloroaniline solution dropwise to the reaction flask containing maleic anhydride solution under stirring, the reaction exotherms and gradually produces a light yellow precipitate , After reacting at room temperature for 1 hour, take a small amount of precipitate, wash with water, dry, 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 changes from orange to Red and black, 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-p-chlorophenylmaleimide (3).

Embodiment 2: the synthesis of 6-bromochromone phenylhydrazone:

Using the method of dehydration reaction of 6-bromochromone with formaldehyde group at the 3 position and phenylhydrazine to generate Schiff base, the specific operation: take 2mmol of phenylhydrazine and add it to a flask containing 10mL of tetrahydrofuran, reflux and stir in a boiling water bath until dissolved, Then slowly add 20 mL of anhydrous ethanol solution in which 2 mmol of 6-bromochromone substituted with formaldehyde group at the 3-position was added dropwise, continue stirring in a boiling water bath under reflux for 1 h, add 10 drops of hydrochloric acid dropwise, and a pale yellow precipitate appears. Continuously reflux and stir in boiling water bath for 5 hours, stop the water bath, add 20mL of distilled water and stir, the light yellow precipitate becomes darker, and phenylhydrazone, a yellow-red needle-like product, is obtained by suction filtration. Rinse with anhydrous ether several times, and dry in vacuum to obtain the product 6-bromochromone phenylhydrazone (4).

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

Add 1mmol N-p-chlorophenyl maleimide and 1.1mmol chromone phenylhydrazone to 20mL ethanol, then add 1.2mL chloramine T, and reflux for 12 hours. The precipitate was filtered, washed, recrystallized with methanol, and dried in vacuo to obtain the product 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-N-p-chlorophenyl-1,6a-di Hydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione (5). The product is yellow crystal, the yield is 54.2%, m.p.173.5-174.5℃.

Confirmation of compound (5) structure: C ₂₆ H ₁₅ BrClN ₃ O ₄

¹ H NMR(DMSO)δ:7.288-7.492(m,12H,Ar-H),6.47(s,1H,C=CH),5.84(d,J=1010Hz,1H),5.33(d,J=1014Hz ,1H),

IR 3457(NC=O), 3085(ArH), 1730(C=O), 1578(C=N), 1292(COC)cm ^-1

m/e: 548.99 (100.0%), 546.99 (77.2%).

_{Anal.calcd.for C26H15BrClN3O4} : C, _{56.92 ;} H, 2.78; N, 7.66.

Application Example 4: MTT method was used to detect the antitumor activity of the test compound (5) on different tumor lines.

The compound (5) prepared in the above example was prepared in different tumor lines [tumor cell Bel-7402 (human liver cancer cell), KB (oral cancer cell), SGC7901 (stomach cancer cell), HO8901 (ovarian cancer cell), HL- 60 (leukemia cells), ECA109 (intestinal cancer cells)] were the experimental objects, and the in vitro inhibitory effect of the test compound (5) on different tumor strains: the experiment used tetramethylazolate microenzyme reaction colorimetry (MTT method) , and the activity was 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) were seeded into 4000 cells/200 μL/well on a 96-well plate, and 2 μL of compound was added to each well, with a final concentration of 12.0 μM, 6.0 μM, 3.0 μM, and 1.5 μM, and the cells were cultured together at 37°C and 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 compounds with high inhibitory activity were selected, and the effects of different concentrations of the compounds on the cycle and apoptosis of human tumor cells were determined.

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, _IC50 values of pyrazole N-p-chlorophenylmaleimide derivatives to 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 which, HL-60 (leukemia cells) has better inhibitory rate and selectivity, and antitumor drugs can be prepared separately , 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)

1.3-(6-Bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c]pyrazole-4,6 (3aH,5H)-diketone, its structural formula is as follows: 2. A 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c]pyrazole -4, the preparation method of 6(3aH, 5H)-diketone is characterized in that, comprises the following steps: reacting maleic anhydride and p-chloroaniline to synthesize N-p-chlorophenylmaleimide, and then using The 6-bromochromone substituted with formaldehyde at the 3-position is dehydrated with phenylhydrazine to synthesize 6-bromochromone phenylhydrazone, and finally the N-p-chlorophenylmaleimide and 6-bromochromone phenylhydrazone A dipolar cycloaddition reaction was carried out to synthesize 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4 -c] pyrazole-4,6(3aH,5H)-dione. 3. a kind of 3-(6-bromo-4-oxygen-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline according to claim 2 [3 ,4-c] the preparation method of pyrazole-4,6 (3aH, 5H)-dione is characterized in that, comprises the following steps: (1), the synthesis of N-p-chlorophenylmaleimide: Dissolve maleic anhydride and p-chloroaniline in acetone solvent, react under stirring, and gradually produce a light yellow precipitate, react at room temperature for 1 hour, take a small amount of precipitate, wash with water, dry, add manganese acetate, triethyl Amine and acetic anhydride, after heating up, the precipitate gradually dissolves, react at 50-60°C for 2 hours, the solution turns from orange to red and black, and new precipitates gradually appear, cool to room temperature, wash the precipitate with water, dry, and recrystallize with acetone Obtain product N-p-chlorophenylmaleimide; (2), the synthesis of 6-bromochromone phenylhydrazone: Take 2mmol of phenylhydrazine and add it to a flask containing 10mL of tetrahydrofuran, stir in a boiling water bath until it dissolves, then slowly add 20mL of anhydrous ethanol solution in which 2mmol of 6-bromochromone substituted with a formaldehyde group at the 3-position is added dropwise, and continue boiling The bath was refluxed and stirred for 1 hour, 10 drops of hydrochloric acid was added dropwise, and a pale yellow precipitate appeared; the boiling water bath was continuously refluxed and stirred for 5 hours, the water bath was stopped, and 20 mL of distilled water was added to stir, the pale yellow precipitate became darker, and 6-bromochromone benzene was obtained by suction filtration Hydrazone, a yellow-red needle-like product, was washed with anhydrous ether several times, and vacuum-dried to obtain the product; (3), 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline[3,4-c]pyrazole Synthesis of -4,6(3aH,5H)-dione: Add 1mmol N-p-chlorophenylmaleimide 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, recrystallized with methanol, and dried in vacuo to obtain the product 3-(6-bromo-4-oxo-4H-chromone)-1-phenyl-N-p-chlorophenyl-1,6a-di Hydropyrroline[3,4-c]pyrazole-4,6(3aH,5H)-dione. 4. a kind of 3-(6-bromo-4-oxygen-4H-chromone)-1-phenyl-5-p-chlorophenyl-1,6a-dihydropyrroline according to claim 3 [3 ,4-c] The preparation method of pyrazole-4,6(3aH,5H)-dione, characterized in that: in the step (1), the light yellow precipitate obtained from the reaction needs to be filtered under reduced pressure. 5. The application of the compound according to claim 1 in the preparation of antitumor drugs.