CN104817535A - Quinolinone derivative, and synthetic method and application thereof - Google Patents

Abstract

The invention discloses a quinolinone derivative and its synthesis method and application. Described quinolinone derivative is 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone, and its synthetic method is: take p-toluidine as raw material, in In the presence of acetic acid or hydrochloric acid, add acetic anhydride to carry out acylation to obtain compound 1; the obtained compound 1 is ring-closed with phosphorus oxychloride to obtain compound 2; the obtained compound 2 is hydrolyzed by adding acid to obtain compound 3; the obtained compound 3 and o-Phenylenediamine condensation reaction, that is. The method of the invention is simple and easy to operate, and the yield is high; the obtained target compound exhibits a certain growth inhibitory activity on various cell lines, among which the activity on human gastric cancer cell MGC-803 is the highest. The structural formula of described quinolinone derivative is shown in following formula (I):

Description

A kind of quinolinone derivative and its synthesis method and application

technical field

The invention relates to the technical field of medicine, in particular to a quinolinone derivative and a synthesis method and application thereof.

Background technique

The 2(1H)-quinolinone structure is an alkaloid that exists widely in natural products like quinoline. Compounds containing the 2(1H)-quinolinone structure have various biological activities. Different substituents can produce broad-spectrum pharmacological activities such as anti-tumor, anti-oxidation, and anti-inflammation. Seeking drug lead compounds with excellent activity and low toxicity is an important method for the development of new anti-tumor drugs. Because 2(1H)-quinolinone has the characteristics of good activity, easy structure modification and low toxicity, it is widely used Applied in the design and screening of anticancer drugs. Some compounds with 2(1H)-quinolinone skeletons have entered the clinic as anti-tumor drugs, for example: Dovitinib is an orally effective small molecule multi-target tyrosine kinase inhibitor, which can directly Acting on tumor cells and blood vessels and stroma that provide nutrition for tumor cells, it exhibits anti-tumor effects through anti-proliferation and anti-vascular survival activities; tipifanib (Tipifanib) belongs to farnesyl transferase inhibitors. Farnesylated proteins prevent the activation of the Ras oncogene, inhibit cell growth, induce apoptosis, and inhibit angiogenesis. So far, there are no related reports on 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone and its synthesis and application.

Contents of the invention

The technical problem to be solved in the present invention is to provide a new 2(1H)-quinolinone derivative, namely 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinoline Ketones, their synthesis and applications.

The present invention relates to a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

The chemical name of the compound represented by the above formula (I) is 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone, and its molecular weight is 275.1.

The synthetic train of thought of the compound shown in above-mentioned formula (I) is: take p-toluidine as raw material, under the condition that acetic acid or hydrochloric acid exist, add acetic anhydride to carry out acylation, obtain acylated product (being compound 1); Gained acylated product Closing the ring with phosphorus oxychloride to obtain a ring-closing product (i.e. compound 2); the resulting ring-closing product is hydrolyzed with acid to obtain a hydrolyzed product (i.e. compound 3); the resulting hydrolyzed product is condensed with o-phenylenediamine to obtain The target product (ie compound 4). Concrete synthetic route is as follows:

Reagents: (a) acetic anhydride, acetic acid or hydrochloric acid; (b) N,N-dimethylformamide, phosphorus oxychloride; (c) acid; (d) o-phenylenediamine, methanol and/or ethanol.

A more specific synthetic method comprises the following steps:

1) Using p-toluidine as a raw material, in the presence of acetic acid or hydrochloric acid, add acetic anhydride to react, after the reaction is completed, adjust the pH value of the system to 6-8, filter the reactant with suction, and recrystallize the filter cake to obtain compound 1 ;

2) The obtained compound 1 was dissolved in N,N-dimethylformamide, phosphorus oxychloride was added to carry out ring closure reaction, the obtained reactant was poured into ice water, and suction filtered to obtain compound 2;

3) The obtained compound 2 is hydrolyzed with acid to obtain compound 3;

4) The obtained compound 3 is condensed with o-phenylenediamine in the presence of methanol and/or ethanol to obtain the target product.

In step 1) of the above-mentioned synthetic method, the concentration of the acetic acid can be 30-90 (v/v)%, the concentration of the hydrochloric acid can be 15-37w/w%, and the consumption of the acetic acid and hydrochloric acid is usually respectively 0.9 to 1.2 times the amount of p-toluidine substance, or greater than 1.2 times the amount of p-toluidine substance. The amount of the acetic anhydride added is usually 0.9 to 1.2 times the amount of the p-toluidine substance, or greater than 1.2 times the amount of the p-toluidine substance. In this step, since the reaction is exothermic, it is preferable to carry out the reaction under ice bath conditions. Whether the reaction is complete can be tracked and detected by thin-layer chromatography (TLC). Usually, it is more appropriate to control the reaction time to 1-4 hours. After the reaction is completed, the pH value of the lye adjustment system is 6-8. The lye can be an aqueous solution of alkaline substances such as sodium acetate, sodium carbonate, sodium phosphate, sodium bicarbonate or potassium carbonate. The lye The concentration is preferably 5-30/w/w%; it is preferably adjusted with an aqueous solution of sodium acetate. The collected filter cake is usually recrystallized using absolute ethanol and/or absolute methanol.

In step 2) of the above synthesis method, the amount of phosphorus oxychloride added is usually 0.9 to 1.2 times the amount of compound 1, or greater than 1.2 times the amount of p-toluidine. The amount of N,N-dimethylformamide can be determined according to needs, specifically, it can be calculated by using 5-11 ml of 10 mmol of compound 1. The ring-closing reaction is usually carried out under heating conditions, preferably at 60-90°C, more preferably at 60-90°C in a reflux device. Whether the reaction is complete can be detected by TLC tracking, usually it is more appropriate to control the reaction time to 8-14h.

In step 3) of the above-mentioned synthetic method, the acid used during hydrolysis can be 30-90 (v/v)% glacial acetic acid, or 2-6 mol/L hydrochloric acid, or 2-6 mol/L sulfuric acid; The amount of acid used for hydrolysis is usually 50-80ml of acid solution per 10mmol of compound 2 for hydrolysis. The hydrolysis is usually carried out under heating conditions, preferably at 60-90°C, more preferably at 60-90°C in a reflux device, and a clear solution can be obtained by reflux at the above temperature conditions. Whether the hydrolysis is complete can be tracked and detected by TLC. Usually, it is more appropriate to control the reaction time to 6-12 hours. After the hydrolysis is complete, the obtained reactant is cooled, and crystals are precipitated, and the precipitated crystals are compound 3.

In step 4) of the above synthesis method, the amount of o-phenylenediamine used is usually 0.9-1.2 times the amount of compound 3, or greater than 1.2 times the amount of p-toluidine. The methanol is 70-100v/v% methanol, and the ethanol is 70-100v/v% ethanol; the amount of methanol and/or ethanol can be determined as needed, specifically 50-80ml of 10mmol compound 3 can be used amount to calculate. The condensation reaction is usually carried out under heating conditions, preferably at 60-90°C, more preferably at 60-90°C in a reflux device. Whether the condensation reaction is complete can be detected by TLC tracking. Usually, it is more appropriate to control the reaction time to 6-12 hours. After the reaction is completed, the obtained reactant is cooled and filtered with suction, and the filter cake is collected as the target product.

The yield of the compound shown in the above (I) synthesized by the synthetic method of the present invention is more than 46%.

The present invention also includes the application of the compound represented by the above (I) or a pharmaceutically acceptable salt thereof in the preparation of antitumor drugs.

Compared with the prior art, the present invention provides a new 2 (1H)-quinolinone derivatives namely 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)- Quinolinone, and its synthetic method and application. The synthesis method obtains the product through four steps of acylation, ring closure, hydrolysis and condensation, the method is simple and easy to operate, and the yield is high. The applicant's research on the in vitro antitumor activity of the compound of the present invention shows that it has a certain proliferation of four cell lines of human cervical cancer cell Hela229, human gastric cancer cell MGC-803, human liver cancer cell line Hep G2 and BEL-7404. Inhibitory activity, among which the activity is the highest for human gastric cancer cell line MGC-803.

Description of drawings

Fig. 1 is the proton nuclear magnetic resonance spectrogram of the final product that the embodiment of the present invention 1 makes;

Fig. 2 is the carbon nuclear magnetic resonance spectrogram of the final product that the embodiment of the present invention 1 makes;

Fig. 3 is the high-resolution mass spectrogram of the final product prepared in Example 1 of the present invention.

Detailed ways

The present invention will be described in further detail below in conjunction with specific examples to better understand the content of the present invention, but the present invention is not limited to the following examples.

Example 1

1) Put 10.7g (0.1mol) of p-toluidine and 50ml of water into a 250ml round bottom flask, add 8ml of concentrated hydrochloric acid and 10.2g (0.1mol) of acetic anhydride dropwise, and react under ice bath for 4 hours, then use 20w/w% The sodium acetate solution was used to adjust the pH of the system to 7, left to stand, suction filtered, and the filter cake was recrystallized with absolute ethanol to obtain compound 1 (white needle-like crystals, 13.3 g, yield 89%).

2) Mix 3.5ml DMF and 17ml POCl ₃ in an ice bath, stir evenly, add 2.24g (15mmol) of compound 1, place the resulting solution in a reflux device, heat to 90°C and reflux for 10 hours, pour into 500ml of ice after cooling In water, the compound 2 was obtained by suction filtration (2.7 g, yield 87%).

3) 2.05g (10mmol) of compound 2 was dissolved in 80ml of 70% glacial acetic acid, the resulting solution was placed in a reflux device, heated to 90°C for reflux for 8 hours, and cooled to obtain compound 3 (yellow needle-like crystals, 1.70g, yield 91 %).

4) Add 1.87g (10mmol) of compound 3 and 1.08g (10mmol) of o-phenylenediamine to 80ml of anhydrous methanol, place the resulting solution in a reflux device, reflux at 90°C for 8 hours, cool, and filter with suction to obtain a yellow Solid (2.48g, 90%).

The resulting yellow solid product is identified:

(1) Proton nuclear magnetic resonance spectrum and carbon spectrum, and their spectrograms are shown in Figures 1 and 2 respectively.

¹ H NMR (500MHz, DMSO-d6) δ12.65(s, 1H), 12.40(s, 1H), 9.00(d, J=1.7Hz, 1H), 7.74–7.70(m, 1H), 7.69(s ,1H),7.68–7.63(m,1H),7.42(d,J＝8.4Hz,1H),7.33(d,J＝8.4Hz,1H),7.22–7.17(m,2H),2.36(s, 3H). ¹³ C NMR (126MHz, DMSO-d6) δ160.79, 147.96, 142.83, 138.90, 136.85, 134.49, 133.15, 131.89, 128.38, 122.41, 122.05, 119.93, 119.19, 118.38, 110.2

(2) Electrospray mass spectrometry, as shown in Figure 3, ESI-MS m/z: 276.1[M+H] ⁺ .

Therefore, it can be determined that the above-mentioned yellow solid product is 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone, and its chemical structural formula is as follows:

Example 2

1) Add 10.7g (0.1mol) p-toluidine and 40ml glacial acetic acid into a 250ml round-bottomed flask, add 10.2g (0.1mol) acetic anhydride dropwise, react under ice bath for 4 hours, then distill off the solvent under reduced pressure, and then The pH of the system was adjusted to 7 with 20w/w% sodium carbonate solution, allowed to stand, filtered with suction, and the filter cake was recrystallized with absolute ethanol to obtain compound 1 (white needle-like crystals, 12.7g, yield 85%).

2) Mix 3.5ml DMF and 17ml POCl ₃ in an ice bath, and add 2.24g (15mmol) of compound 1 after uniform stirring. The resulting solution is placed in a reflux device, heated to 70°C and refluxed for 14 hours, and poured into 500ml ice In water, the compound 2 was obtained by suction filtration (2.5 g, yield 80%).

3) 2.05g (10mmol) of compound 2 was dissolved in 80ml of 90% glacial acetic acid, the resulting solution was placed in a reflux device, heated to 60°C for reflux for 10 hours, and cooled to obtain compound 3 (yellow needle-like crystals, 1.50g, yield 80 %).

4) Add 1.87g (10mmol) of compound 3 and 1.08g (10mmol) of o-phenylenediamine into 80ml of 80v/v% methanol, place the resulting solution in a reflux device, heat to reflux at 90°C for 8 hours, cool, and filter with suction to obtain Yellow solid (2.48 g, 90%).

The obtained yellow solid product was identified as 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone.

Example 3:

1) Put 10.7g (0.1mol) of p-toluidine and 50ml of water into a 250ml round bottom flask, add 8ml of concentrated hydrochloric acid and 10.2g (0.1mol) of acetic anhydride dropwise, and react under ice bath for 4 hours, then use 20w/w% The sodium acetate solution was used to adjust the pH of the system to 6, left to stand, suction filtered, and the filter cake was recrystallized with absolute ethanol to obtain white needle-like crystal 1 (13.3 g, yield 89%).

2) Mix 3.5ml DMF and 17ml POCl ₃ in an ice bath, stir evenly and add 2.24g (15mmol) of compound 1, place the resulting solution in a reflux device, heat to 60°C and reflux for 10 hours, pour into 500ml of ice after cooling In water, the compound 2 was obtained by suction filtration (2.4 g, yield 77%).

3) 2.05g (10mmol) of compound 2 was dissolved in 80ml of 4M hydrochloric acid aqueous solution, and the resulting solution was placed in a reflux device, heated to 90°C for reflux for 10 hours, and cooled to obtain compound 3 (yellow needle-like crystals, 1.59g, yield 85% ).

4) Add 1.87g (10mmol) of compound 3 and 1.08g (10mmol) of o-phenylenediamine into 100ml of absolute ethanol, place the resulting solution in a reflux device, heat to reflux at 80°C for 6 hours, cool, and filter with suction to obtain a yellow Solid (2.4 g, 87%).

The obtained yellow solid product was identified as 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone.

Example 4:

1) Put 10.7g (0.1mol) of p-toluidine and 50ml of water into a 250ml round bottom flask, add 8ml of concentrated hydrochloric acid and 10.2g (0.1mol) of acetic anhydride dropwise, and react under ice bath for 4 hours, then use 20w/w% The sodium acetate solution was used to adjust the pH of the system to 7.5, left to stand, suction filtered, and the filter cake was recrystallized from anhydrous methanol to obtain compound 1 (white needle-like crystals, 13.3 g, yield 89%).

2) Mix 3.5ml DMF and 17ml POCl ₃ under ice bath, add 2.24g (15mmol) of compound 1 after uniform stirring, place the resulting solution in a reflux device, heat to 90°C and reflux for 8 hours, pour into 500ml of ice after cooling In water, the compound 2 was obtained by suction filtration (2.6 g, yield 83%).

3) 2.05g (10mmol) of compound 2 was dissolved in 80ml of 40% glacial acetic acid, and the resulting solution was placed in a reflux device, heated to 90°C for reflux for 6 hours, and cooled to obtain compound 3 (yellow needle-like crystals, 1.4g, yield 75 %).

4) Add 1.87g (10mmol) of compound 3 and 1.08g (10mmol) of o-phenylenediamine into 80ml of 90v/v% ethanol, place the resulting solution in a reflux device, heat to reflux at 60°C for 12 hours, cool, and filter with suction to obtain Yellow solid (2.3 g, 83%).

The obtained yellow solid product was identified as 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone.

The applicant has carried out in vitro anti-tumor activity experiment on 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone described in the present invention, specifically as follows:

1. Cell lines and cell culture

In this experiment, four cell lines including human cervical cancer cell Hela229, human gastric cancer cell line MGC-803, human liver cancer cell line Hep G2 and BEL-7404 were selected.

All tumor cell lines were cultured in RPMI-1640 medium containing 10wt% calf blood, 100U/mL penicillin, and 100U/mL streptomycin in an incubator at 37°C with a volume concentration of 5% _CO2 ; The cell lines were cultured in DMEM medium containing 10wt% calf blood, 100U/mL penicillin, and 100U/mL streptomycin.

2. Preparation of test compounds

The 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone used is obtained by purifying the product obtained by the method described in Example 1 of the present invention through column chromatography, Purity ≥ 95%, its DMSO stock solution (concentration is 0.001mol/L) was diluted into five concentration gradients successively through RMPI1640 medium, respectively 40, 20, 10, 5, 2.5 μmol/L, wherein the cosolvent DMSO was finally Concentration ≤ 1%. First test the inhibitory rate of 20 μmol/L target product on the proliferation of tumor cells, which is regarded as the result of the preliminary screening; then test the degree of inhibition of the target product on the proliferation of various tumor cells at different gradient concentrations, and use it to fit and calculate the half inhibitory concentration. That is the _IC50 value.

3. Cell growth inhibition test (MTT method)

(1) Take the tumor cells in the logarithmic growth phase, digest them with trypsin, prepare a cell suspension with a concentration of 5000 cells/mL with a culture medium containing 10% calf serum, inoculate 190 μL per well in 96-well culture In the plate, make the cell density to be tested to 1000-10000/well (the edge wells are filled with sterile PBS);

(2) 5% CO ₂ , incubate at 37°C for 24 hours, until the cell monolayer covers the bottom of the well, add 10 μL of drug with a certain concentration gradient to each well, and set 4 replicate wells for each concentration gradient;

(3) 5% CO ₂ , incubate at 37°C for 48 hours, observe under an inverted microscope;

(4) Add 10 μL of MTT solution (5 mg/mL PBS, ie 0.5% MTT) to each well, and continue to incubate for 4 h;

(5) Terminate the culture, carefully suck out the culture medium in the wells, add 150 μL DMSO to each well to fully dissolve the formazan precipitate, mix well with a shaker, and measure the light in each well in a microplate reader with a wavelength of 570nm and a reference wavelength of 450nm. density value;

(6) At the same time, set zero adjustment wells (medium, MTT, DMSO) and control wells (cells, drug dissolution medium with the same concentration, culture medium, MTT, DMSO).

(7) According to the measured optical density value (OD value), the number of living cells is judged. The larger the OD value, the stronger the cell activity.

Use the formula:

Tumor cell growth inhibition rate

Calculate the inhibition rate of the compound on tumor cell growth. The test results are shown in Table 1 below.

Table 1: The growth inhibitory rate (%) of compound of the present invention to different tumor cell lines when 20 μ mol/L

For the cell lines with an inhibition rate exceeding 50% at the initial screening concentration, the data of the inhibition rate of five concentration gradients were further fitted by SPSS software, and the half inhibitory concentration ( _IC50 value, unit μmol) of the product to different tumor lines was obtained. /L), the _IC50 values of the compounds of the present invention for different cell lines are shown in Table 2.

Table 2: _IC50 values (μM) of each cell line of the compounds of the present invention

From the in vitro antitumor activity test results, 3-(1H-benzimidazol-2-yl)-6-methyl-2(1H)-quinolinone of the present invention is effective on human cervical cancer cells Hela229, human Gastric cancer cell line MGC-803, human liver cancer cell line Hep G2 and BEL-7404 four cell lines all showed certain anti-proliferation activity, among which the activity was the highest for human gastric cancer cell line MGC-803.

Claims (10)

1. The compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof: 2. the compound method of the described compound of claim 1 is characterized in that: take p-toluidine as raw material, under the condition that acetic acid or hydrochloric acid exist, add acetic anhydride and carry out acidification, obtain compound 1; Gained compound 1 is with phosphorus oxychloride The ring is closed to obtain compound 2; the obtained compound 2 is hydrolyzed with acid to obtain compound 3; the obtained compound 3 is condensed with o-phenylenediamine to obtain the target product. 3. the compound method of the described compound of claim 2 is characterized in that: comprise the following steps: 1) Using p-toluidine as a raw material, in the presence of acetic acid or hydrochloric acid, add acetic anhydride to react, after the reaction is completed, adjust the pH value of the system to 6-8, filter the reactant with suction, and recrystallize the filter cake to obtain compound 1 ; 2) The obtained compound 1 was dissolved in N,N-dimethylformamide, phosphorus oxychloride was added to carry out ring closure reaction, the obtained reactant was poured into ice water, and suction filtered to obtain compound 2; 3) The obtained compound 2 is hydrolyzed with acid to obtain compound 3; 4) The obtained compound 3 is condensed with o-phenylenediamine in the presence of methanol and/or ethanol to obtain the target product. 4. synthetic method according to claim 3 is characterized in that: in step 1), reaction is carried out under ice-bath condition. 5. The synthesis method according to claim 3, characterized in that: in step 1), the pH value of the system is adjusted to 6-8 with lye. 6. The synthesis method according to claim 3, characterized in that: in step 2), the ring closure reaction is carried out at 60-90°C. 7. The synthetic method according to claim 3, characterized in that: in step 3), the acid used for hydrolysis is 30-90 (v/v)% acetic acid, or 2-6mol/L hydrochloric acid, or It is 2~6mol/L sulfuric acid. 8. The synthesis method according to claim 3, characterized in that in step 3), the hydrolysis is carried out at 60-90°C. 9. The synthesis method according to claim 3, characterized in that: in step 4), the condensation reaction is carried out at 60-90°C. 10. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of antitumor drugs.