CN111606848A - A kind of preparation method of fluorodiphenyl substituted pyridine compounds - Google Patents

Abstract

The invention discloses a preparation method of a fluorodiphenyl substituted pyridine compound, which comprises the following steps: s1 adding NKC-9 catalyst, 4-hydroxyacetophenone, 2-trifluoromethylbenzaldehyde and appropriate amount of CHCl3 into a round bottom flask, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, filtering out the catalyst after the reaction is finished, washing the catalyst by using a small amount of chloroform, concentrating the filtrate to obtain a crude product, and recrystallizing by using 95% ethanol to obtain a chalcone precursor compound; adding the product of the last step, malononitrile and ammonium acetate into a round-bottom flask according to a molar ratio of 1:1: 3. The compound of the invention takes substituted acetophenone, substituted benzaldehyde, malononitrile, ethyl cyanoacetate and ammonium acetate as raw materials, a one-pot method is adopted to synthesize the 4, 6-diphenyl-3 cyanopyridine compound, and MTT (methyl thiazolyl tetrazolium) method tests show that the compound has strong inhibition effect on the proliferation of glioma U251 and has good application prospect in screening antitumor drugs.

Description

A kind of preparation method of fluorodiphenyl substituted pyridine compounds

technical field

The invention relates to the technical field of compound preparation, in particular to a preparation method of a fluorodiphenyl substituted pyridine compound.

Background technique

The coordination and weak interaction between organic ligands and metal ions to construct coordination polymers with novel structures and unique properties has become a hot research topic in the fields of coordination chemistry, supramolecular chemistry, crystal engineering, and materials chemistry. Designing and synthesizing new coordination polymer materials by using the unique bonding method of coordination chemistry has become a common development direction of coordination chemistry and materials chemistry.

The compound of the present invention uses substituted acetophenone, substituted benzaldehyde, malononitrile, ethyl cyanoacetate, and ammonium acetate as raw materials, and adopts a one-pot method to synthesize 4,6-diphenyl-3 cyanopyridine compounds. The MTT method test showed that the drug has a strong inhibitory effect on the proliferation of glioma U251, and has a good application prospect in the screening of anti-tumor drugs.

SUMMARY OF THE INVENTION

Based on the technical problems existing in the background art, the present invention proposes a preparation method of fluorodiphenyl substituted pyridine compounds.

The preparation method of a kind of fluorodiphenyl substituted pyridine compound proposed by the present invention comprises the following steps:

S1 Add NKC-9 catalyst, 4-hydroxyacetophenone, 2-trifluoromethylbenzaldehyde to a round-bottomed flask, add an appropriate amount of CHCl3, heat to reflux under nitrogen protection, follow the reaction by TLC, after the reaction is over, filter out the catalyst Wash with a small amount of chloroform, and concentrate the filtrate to obtain a crude product, which is then recrystallized with 95% ethanol to obtain a chalcone precursor compound;

The product of the previous step, malononitrile and ammonium acetate were added to a round-bottomed flask according to a molar ratio of 1:1:3, an appropriate amount of absolute ethanol was added, heated to reflux under nitrogen protection, and the reaction was followed by TLC. After the reaction was completed, concentrated to the reaction mixture, extract the reaction mixture with ethyl acetate:water=1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product, which is named 2-amino-6 -(4-Hydroxyphenyl)-4(2-trifluoromethylphenyl)-3-cyanopyridine, referred to as 1a;

S2 Add NKC-9 catalyst, 4-hydroxyacetophenone, 3-trifluoromethylbenzaldehyde to the round-bottomed flask, add an appropriate amount of CHCl3, heat to reflux under nitrogen protection, follow the reaction by TLC, after the reaction is over, filter out the catalyst Wash with a small amount of chloroform, and concentrate the filtrate to obtain a crude product, which is then recrystallized with 95% ethanol to obtain a chalcone precursor compound;

The product of the previous step, malononitrile and ammonium acetate were added to a round-bottomed flask in a molar ratio of 1:1:3, an appropriate amount of anhydrous ethanol was added, heated to reflux under nitrogen protection, and the reaction was followed by TLC. After the reaction was completed, concentrated to the reaction mixture, extract the reaction mixture with ethyl acetate:water=1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product, which is named 2-amino-6 -(4-Hydroxyphenyl)-4(3-trifluoromethylphenyl)-3-cyanopyridine, referred to as 2a;

S3 Add NKC-9 catalyst, 2,4-difluoroacetophenone, 4-hydroxybenzaldehyde to a round-bottomed flask, add an appropriate amount of CHCl3, heat to reflux under nitrogen protection, follow the reaction by TLC, after the reaction is over, filter out the catalyst Wash with a small amount of chloroform, and concentrate the filtrate to obtain a crude product, which is then recrystallized with 95% ethanol to obtain a chalcone precursor compound;

The product of the previous step, malononitrile and ammonium acetate were added to a round-bottomed flask according to a molar ratio of 1:1:3, an appropriate amount of absolute ethanol was added, heated to reflux under nitrogen protection, and the reaction was followed by TLC. After the reaction was completed, concentrated to the reaction mixture, extract the reaction mixture with ethyl acetate:water=1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product, which is named 2-amino-6 -(2,4-Difluorophenyl)-4-(4-hydroxymethylphenyl)-3-cyanopyridine, referred to as 3a;

S4 Add NKC-9 catalyst, 2,4-difluoroacetophenone, 4-hydroxybenzaldehyde to a round-bottomed flask, add an appropriate amount of CHCl3, heat to reflux under nitrogen protection, follow the reaction by TLC, after the reaction is over, filter out the catalyst Wash with a small amount of chloroform, and concentrate the filtrate to obtain a crude product, which is then recrystallized with 95% ethanol to obtain a chalcone precursor compound; the product of the previous step, ethyl cyanoacetate and ammonium acetate are added in a molar ratio of 1:1:3 Into a round-bottomed flask, add an appropriate amount of anhydrous ethanol, heat to reflux under nitrogen protection, follow the reaction by TLC. After the reaction is completed, concentrate the reaction mixture, extract the reaction mixture with ethyl acetate: water = 1:1, Evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product, whose name is 2-amino-6-(2,4-difluorophenyl)-4-(4-hydroxymethylphenyl) )-2oxo-1,2-dihydro-3 cyanopyridine, referred to as 4a.

Preferably, the NKC-9 catalyst is a macroporous strongly acidic styrene-based cation exchange resin.

In the present invention, for the preparation method of a fluorodiphenyl substituted pyridine compound, the compound of the present invention uses substituted acetophenone, substituted benzaldehyde, malononitrile, ethyl cyanoacetate and ammonium acetate as raw materials, and adopts a The pot method was used to synthesize 4,6-diphenyl-3 cyanopyridine compounds. After the MTT method, it was found that the drug has a strong inhibitory effect on the proliferation of glioma U251, and it has a good effect in screening anti-tumor drugs. application prospects.

Detailed ways

The technical solutions in the embodiments of the present invention will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments.

Example 1

The synthetic route of 2-amino-6-(4-hydroxyphenyl)-4(2-trifluoromethylphenyl)-3-cyanopyridine is as follows:

Weigh 4-hydroxyacetophenone (408mg, 3mmol), 2-trifluoromethylbenzaldehyde (522mg3mmol), add 0.4g NKC-9 catalyst to the round-bottomed flask, add 5ml chloroform to dissolve, rush into nitrogen protection, reflux reaction , TLC monitors the reaction progress, after the reaction is complete, cool to room temperature, filter out the NKC-9 catalyst, concentrate the filtrate to obtain the crude product of the intermediate, and recrystallize the crude product of the intermediate with 95% ethanol to obtain the crude product of the intermediate. Pure.

Weigh intermediate 3mmol, malononitrile (198mg, 3mmol), ammonium acetate (514mg, 9mmol) were added to the round-bottomed flask, 20ml absolute ethanol was added, nitrogen protection was added, and the reaction was refluxed for 8h. After the reaction is complete, concentrate to obtain the reaction mixture, extract the reaction mixture with ethyl acetate: water = 1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product.

Its name is 2-amino-6-(4-hydroxyphenyl)-4(2-trifluoromethylphenyl)-3-cyanopyridine, referred to as 1a, and its structural formula is as follows:

Product 1a is a pale yellow powder solid (yield 43%), 1HNMR (400MHz, DMSO-d6)δ9.97(s,1H), 8.13–8.03(m,0H), 8.00–7.88 (m,2H), 7.82(td, J=7.8, 1.3Hz, 1H), 7.78-7.68(m, 1H), 7.64(s, 0H), 7.57-7.49(m, 1H), 7.08(s, 1H), 7.01(s, 1H), 6.94–6.82(m, 2H).13CNMR(101MHz,DMSO-d6)δ160.17, 158.42,153.36,136.55,133.12,131.36,130.01,129.39,128.64, 128.47,126.72,116.74,115 87.60.

Example 2

2-Amino-6-(4-hydroxyphenyl)-4(3-trifluoromethylphenyl)-3-cyanopyridine

Its synthetic route is as follows:

4-Hydroxyacetophenone (408mg, 3mmol), 3-trifluoromethylbenzaldehyde (522mg3mmol), 0.4g NKC-9 catalyst were weighed into the round-bottomed flask, 5ml of chloroform was added to dissolve, flushed into nitrogen protection, and refluxed for reaction , TLC monitors the reaction progress, after the reaction is complete, cool to room temperature, filter out the NKC-9 catalyst, concentrate the filtrate to obtain the crude product of the intermediate, and recrystallize the crude product of the intermediate with 95% ethanol to obtain the crude product of the intermediate. Pure.

Weigh intermediate 3mmol, malononitrile (198mg, 3mmol), ammonium acetate (514mg, 9mmol) were added to the round-bottomed flask, 20ml absolute ethanol was added, nitrogen protection was added, and the reaction was refluxed for 8h. After the reaction is complete, concentrate to obtain the reaction mixture, extract the reaction mixture with ethyl acetate: water = 1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product.

Its name is 2-amino-6-(4-hydroxyphenyl)-4(3-trifluoromethylphenyl)-3-cyanopyridine, referred to as 2a, and its structural formula is as follows:

Product 2a is a pale yellow solid (yield 42%), 1HNMR (400MHz, DMSO-d6)δ9.97(s,1H), 8.08–7.96(m,4H), 7.94–7.88 (m,1H), 7.80 (t, J=7.8 Hz, 1H), 7.27 (s, 1H), 7.01 (s, 2H), 6.90–6.85 (m, 2H).

13CNMR(101MHz,DMSO-d6)δ161.20,160.15,159.38,138.60,133.04,130.32,129.56,128.87,128.67,126.57(d,J=3.9Hz),125.53(d,J=4.0Hz),117.54,115 108.74, 85.62.

Example three

2-amino-6-(2,4-difluorophenyl)-4-(4-hydroxymethylphenyl)-3-cyanopyridine;

Its synthetic route is as follows:

Weigh 2,4-difluoroacetophenone (408mg, 3mmol), 4-hydroxybenzaldehyde (522mg3mmol), add 0.4g NKC-9 catalyst to the round bottom flask, add 5ml chloroform to dissolve, flush with nitrogen protection, reflux reaction , TLC monitors the reaction progress, after the reaction is complete, cool to room temperature, filter out the NKC-9 catalyst, concentrate the filtrate to obtain the crude product of the intermediate, and recrystallize the crude product of the intermediate with 95% ethanol to obtain the crude product of the intermediate. Pure.

Weigh intermediate 3mmol, malononitrile (198mg, 3mmol), ammonium acetate (514mg, 9mmol) were added to the round-bottomed flask, 20ml absolute ethanol was added, nitrogen protection was added, and the reaction was refluxed for 8h. After the reaction is complete, concentrate to obtain the reaction mixture, extract the reaction mixture with ethyl acetate: water = 1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product.

Its name is 2-amino-6-(2,4-difluorophenyl)-4-(4-hydroxymethylphenyl)-3-cyanopyridine, referred to as 3a, and its structural formula is as follows:

Product 3a is a white solid (yield 40%), 1H NMR (400MHz, DMSO-d6) δ10.00 (d, J=1.3Hz, 1H), 8.06-7.80 (m, 1H), 7.51 (dd, J= 8.4, 1.6Hz, 2H), 7.39 (ddd, J=11.7, 9.2, 2.4Hz, 1H), 7.26 (td, J=8.5, 2.3Hz, 1H), 7.10–6.98 (m, 3H), 6.99– 6.87 (m,2H).13CNMR(101MHz,DMSO-d6)δ161.38,159.52,155.10,154.50,133.52–131.96(m),130.24,127.60,124.05–122.29(m),117.58,116.08,113.30–112.19(m) , 105.22(t, J=26.8Hz), 87.21.

Example 4

2-Amino-6-(2,4-difluorophenyl)-4-(4-hydroxymethylphenyl)-2oxo-1,2-dihydro-3cyanopyridine

The synthetic route is as follows:

Weigh 2,4-difluoroacetophenone (408mg, 3mmol), 4-hydroxybenzaldehyde (522mg3mmol), add 0.4g NKC-9 catalyst to the round bottom flask, add 5ml chloroform to dissolve, flush with nitrogen protection, reflux reaction , TLC monitors the reaction progress, after the reaction is complete, cool to room temperature, filter out the NKC-9 catalyst, concentrate the filtrate to obtain the crude product of the intermediate, and recrystallize the crude product of the intermediate with 95% ethanol to obtain the crude product of the intermediate. Pure

Weighing intermediate 3mmol, ethyl cyanoacetate (340mg, 3mmol), ammonium acetate (514mg, 9mmol) were added to the round-bottomed flask, 20ml of absolute ethanol was added, nitrogen protection was added, the reaction was refluxed for 8h, and the reaction progress was monitored by TLC, After the reaction is complete, concentrate to obtain the reacted mixture, extract the reacted mixture with ethyl acetate:water=1:1, evaporate the ethyl acetate to dryness to obtain the crude product, and recrystallize the crude product in ethanol to obtain the target product.

Its name is 2-amino-6-(2,4-difluorophenyl)-4-(4-hydroxymethylphenyl)-2oxo-1,2-dihydro-3 cyanopyridine, referred to as 4a , and its structure is as follows:

Product 4a is a white solid (yield 45%), nuclear magnetic 1HNMR (400MHz, DMSO-d6) δ12.77 (s, 1H), 10.18 (s, 1H), 7.81 (td, J=8.6, 6.4Hz, 1H) , 7.71–7.58(m, 2H), 7.49(ddd, J=11.4, 9.3, 2.5Hz, 1H), 7.28(td, J=8.5, 2.6Hz, 1H), 7.05–6.89(m, 2H), 6.63 (s, 1H).13CNMR(101MHz,DMSO-d6)δ165.38(d,J=12.7Hz), 162.89(d,J=12.4Hz),162.19,161.36(d,J=12.9Hz),160.40, 158.84(d, J＝13.0Hz), 132.95(dd, J＝10.4, 3.5Hz), 130.62, 126.49, 118.42, 117.23, 116.09, 112.69(dd, J＝21.7, 3.5Hz), 108.68, 105.36(t, J=26.2Hz), 98.61.

Antitumor effect of biphenylpyridine compounds 1-4;

Select u251 as the target cell, use 10-hydroxycamptothecin as the positive drug, and measure the antiproliferative effect of the cells by MTT colorimetry. Take 0.25% trypsinized monolayer cultured u251 cells, adjust with DEME medium containing 10% fetal bovine serum to make a single cell suspension of 1*105 per ml, and inoculate 100ul per well in a 96-well plate.

The culture plate was placed in a carbon dioxide CO2 incubator, and incubated at 37°C under 5% CO2 saturated humidity conditions for 24 hours. After the cells adhered, the old medium was sucked out, and 40uM, 20uM, 10uM, 5uM, 2.5uM of the drug to be tested were added respectively. and 5% fetal bovine serum (FBS), 5 replicate wells were set for each concentration setting, and the experiment was set up as a blank group, and continued to be placed in a CO2 incubator, 37°C, and 5% CO2-saturated humidity for 48 hours; then aspirated and cultured base, add 20ul MTT (5mg/L) to each well, continue to put it in a CO2 incubator, incubate at 37°C, 5% CO2-saturated humidity for 1 h; add 100ul DMSO, shake to dissolve the formazan precipitate, and measure it on an enzyme-linked immunosorbent assay (ELISA) Measure the absorbance (A) value of each well at 490 nm, according to the formula: inhibition rate=(average A value of negative control group-average A value of administration group)/(average A value of control group-A value of blank control group)*100%.

Calculate the rate of inhibition of cells by compounds

The table shows the anti-tumor proliferation activity of 4,6-diphenyl-3 cyanopyridine compounds

compound u251IC50(umol/l) 10-Hydroxycamptothecin 55.35 1a 23.05 2a 17.64 3a 6.03 4a 12.01

The above description is only a preferred embodiment of the present invention, but the protection scope of the present invention is not limited to this. The equivalent replacement or change of the inventive concept thereof shall be included within the protection scope of the present invention.

Claims (2)

1. A preparation method of a fluorodiphenyl substituted pyridine compound is characterized by comprising the following steps:

s1 adding NKC-9 catalyst, 4-hydroxyacetophenone, 2-trifluoromethylbenzaldehyde and appropriate amount of CHCl3 into a round bottom flask, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, filtering out the catalyst after the reaction is finished, washing the catalyst by using a small amount of chloroform, concentrating the filtrate to obtain a crude product, and recrystallizing by using 95% ethanol to obtain a chalcone precursor compound;

adding the product of the last step, malononitrile and ammonium acetate into a round-bottom flask according to a molar ratio of 1:1:3, adding a proper amount of absolute ethanol, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, concentrating to obtain a post-reaction mixture after the reaction is finished, and adding ethyl acetate: extracting the reacted mixture with 1:1, evaporating ethyl acetate to dryness to obtain a crude product, and recrystallizing the crude product in ethanol to obtain a target product, wherein the name of the target product is 2-amino-6- (4-hydroxyphenyl) -4 (2-trifluoromethylphenyl) -3-cyanopyridine, which is called 1a for short;

s2 adding NKC-9 catalyst, 4-hydroxyacetophenone, 3-trifluoromethylbenzaldehyde into a round-bottom flask, adding a proper amount of CHCl3, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, filtering out the catalyst after the reaction is finished, washing the catalyst by using a small amount of chloroform, concentrating the filtrate to obtain a crude product, and recrystallizing by using 95% ethanol to obtain a chalcone precursor compound;

adding the product of the last step, malononitrile and ammonium acetate into a round-bottom flask according to a molar ratio of 1:1:3, adding a proper amount of absolute ethanol, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, concentrating to obtain a post-reaction mixture after the reaction is finished, and adding ethyl acetate: extracting the reacted mixture with water at a ratio of 1:1, evaporating ethyl acetate to dryness to obtain a crude product, and recrystallizing the crude product in ethanol to obtain a target product, wherein the name of the target product is 2-amino-6- (4-hydroxyphenyl) -4 (3-trifluoromethylphenyl) -3-cyanopyridine, which is called 2a for short;

s3 adding an NKC-9 catalyst, 2, 4-difluoroacetophenone, 4-hydroxybenzaldehyde and a proper amount of CHCl3 into a round-bottom flask, heating and refluxing under the protection of nitrogen, tracking and reacting by TLC, filtering out the catalyst after the reaction is finished, washing the catalyst by using a small amount of chloroform, concentrating the filtrate to obtain a crude product, and recrystallizing by using 95% ethanol to obtain a chalcone precursor compound;

adding the product of the last step, malononitrile and ammonium acetate into a round-bottom flask according to a molar ratio of 1:1:3, adding a proper amount of absolute ethanol, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, concentrating to obtain a post-reaction mixture after the reaction is finished, and adding ethyl acetate: extracting the reacted mixture with 1:1, evaporating ethyl acetate to dryness to obtain a crude product, and recrystallizing the crude product in ethanol to obtain a target product, wherein the name of the target product is 2-amino-6- (2, 4-difluorophenyl) -4- (4-hydroxymethylphenyl) -3-cyanopyridine, which is called 3a for short;

s4 adding an NKC-9 catalyst, 2, 4-difluoroacetophenone, 4-hydroxybenzaldehyde and a proper amount of CHCl3 into a round-bottom flask, heating and refluxing under the protection of nitrogen, tracking and reacting by TLC, filtering out the catalyst after the reaction is finished, washing the catalyst by using a small amount of chloroform, concentrating the filtrate to obtain a crude product, and recrystallizing by using 95% ethanol to obtain a chalcone precursor compound; adding the product of the previous step, ethyl cyanoacetate and ammonium acetate into a round-bottom flask according to a molar ratio of 1:1:3, adding a proper amount of absolute ethyl alcohol, heating and refluxing under the protection of nitrogen, tracking the reaction by TLC, concentrating to obtain a post-reaction mixture, and adding ethyl acetate: extracting the reacted mixture with 1:1 of water, evaporating ethyl acetate to dryness to obtain a crude product, and recrystallizing the crude product in ethanol to obtain a target product, wherein the name of the target product is 2-amino-6- (2, 4-difluorophenyl) -4- (4-hydroxymethylphenyl) -2 oxo-1, 2-dihydro-3 cyanopyridine, which is 4a for short.

2. The method for preparing a fluorodiphenyl-substituted pyridine compound according to claim 1, wherein said NKC-9 catalyst is a macroporous strongly acidic styrene cation exchange resin.