CN103819408B - Nitro imidazole derivatives and its production and use - Google Patents

Abstract

The present invention relates to a kind of nitro imidazole derivatives and its production and use, its general formula of molecular structure is as follows:

Description

Nitroimidazole derivatives and their preparation and use

technical field

The invention relates to a nitroimidazole derivative, a preparation method and application thereof, and belongs to the technical field of organic synthesis.

Background technique

Nitroimidazoles have a unique killing effect on anaerobic bacteria and protozoa, and have been widely used in various clinical fields in combination with other antibiotics. Representative drugs for its clinical application include: Metronidazole (MNZ), Tinidazole (Tinidazole, TNZ), Dimetridazole (DMZ), Secnidazole (Secnidazole, SCZ). Based on the excellent antibacterial activity of nitroimidazoles, they are often used as intermediates for the design and synthesis of new drugs.

Contents of the invention

The object of the present invention is to provide a nitroimidazole derivative and its preparation method and application. The derivative has good anticancer activity and can be used in the preparation of anticancer drugs.

Nitroimidazole derivatives of the present invention, its molecular structure general formula is as follows:

in:

When R1=R2=R4=R5=H, R3 is Cl, Br, CH ₃ , OCH ₃ , NO ₂ or H;

When R1=R2=R3=R5=H, R4 is F, Cl, Br or CH ₃ ;

When R1=R2=R3=R4=H, R5 is F, Cl, Br, CH ₃ , OCH ₃ or NO ₂ ;

When R1=N, R1=R4=R5=H, R3 is Cl;

When R1=N, R2=R3=R4=H, R5 is Cl, CH ₃ or OCH ₃ ;

When R2=N, R1=R3=R4=R5=H.

The preparation method of nitroimidazole derivative, comprises the steps:

(1) Dissolve 2,4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, and tetrabutylammonium bromide in absolute ethanol, react at 85°C for 10 minutes, and then wait The solution is filtered after cooling, and the filtered solid is washed twice with distilled water, filtered, and recrystallized to obtain product a;

(2) Mix product a, hydroxylamine hydrochloride and potassium carbonate, dissolve in absolute ethanol, react at 90°C for 2 to 4 hours, then evaporate the solvent to dryness, extract with water and ethyl acetate, keep the organic layer, anhydrous sodium sulfate Evaporate to dryness after drying to obtain product b;

(3) React the product b with substituted benzoic acid or substituted nicotinic acid, react at room temperature for 6-12 hours, evaporate the solvent to dryness after the reaction is complete, extract with water and ethyl acetate, keep the organic solvent layer, dry with anhydrous sodium sulfate and evaporate to dryness A nitroimidazole derivative is obtained.

The molar ratio of 2,4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate and tetrabutylammonium bromide is 1:1:1-3:0.05-1, the preferred molar ratio It is 1:1:2:0.5.

The molar ratio of product a, hydroxylamine hydrochloride and potassium carbonate is 1:1-10:1-3, preferably 1:6:2.

The molar ratio of product b to substituted benzoic acid or substituted nicotinic acid is 1:1-1.5, preferably 1:1.

In step (3), the reaction solvent is dichloromethane, the catalyst is 6-chloro-1-hydroxybenzotriazole (HOBT), and the binder is 1-ethyl-(3-dimethylaminopropyl) Carbodiimide hydrochloride (EDC.HCl).

The molar ratio of product b, HOBT and EDC.HCl is 1:0.05-1:1-2.

The synthetic route is as follows:

The present invention introduces a nitroimidazole functional group on the basis of 2,4'-dibromoacetophenone compound, then introduces a hydroxylamine functional group at the keto group position and further introduces a heterocyclic ring at this position. Many of the compounds thus obtained have good Bioactive functional groups are expected to obtain better anticancer activity.

Compared with the prior art, the beneficial effects of the present invention are as follows:

The nitroimidazole derivatives of the invention not only have better anticancer activity, but also can be used in the preparation of anticancer drugs, and the preparation method is simple and easy to operate. The required raw materials are cheap and easy to obtain, the yield is high, the post-treatment purification method is simple, and the method is suitable for industrial production.

Detailed ways

The present invention will be further described below in conjunction with the examples.

The chemical structural formula of the nitroimidazole derivatives in Examples 1-21 is as follows:

Example 1

When R1=R2=R4=R5=H, when R3 is Cl, the preparation steps are as follows:

(1) Dissolve 2,4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, and tetrabutylammonium bromide in absolute ethanol, and react at 85°C for 10 minutes, 2, The molar ratio of 4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, and tetrabutylammonium bromide is 1:1:3:1, and then wait for the solution to cool and then filter it. The solid obtained was washed twice with distilled water, filtered, and recrystallized to obtain product a;

(2) Mix product a with hydroxylamine hydrochloride and potassium carbonate at a molar ratio of 1:8:1, dissolve in absolute ethanol, and react at 90°C for 4 hours. Then evaporate the solvent to dryness, extract with water and ethyl acetate, keep the organic layer, dry over anhydrous sodium sulfate and evaporate to dryness to obtain product b;

(3) React the product b with o-chlorobenzoic acid in an equimolar ratio of 1:1.5, and react at room temperature for 12 hours. After the reaction is complete, evaporate the solvent to dryness, extract with water and ethyl acetate, keep the organic solvent layer, and dry it with anhydrous sodium sulfate Evaporate to dryness to obtain nitroimidazole derivatives. The reaction solvent is dichloromethane, the catalyst is HOBT, and the binder is EDC.HCl, wherein the molar ratio of product b, HOBT and EDC.HCl is 1:0.05:1.

The product is a white powder with a yield of 60%, m.p.171.2-171.3oC.

Example 2

When R1=N, R2=R3=R4=H, and R5 is Cl, the preparation method is as follows:

(1) Dissolve 2,4’-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, and tetrabutylammonium bromide in absolute ethanol, and react at 85°C for 10 minutes. The molar ratio of 2,4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, and tetrabutylammonium bromide is 1:1:2:0.5, and then wait for the solution to cool and filter , the filtered solid was washed twice with distilled water, filtered, and recrystallized to obtain product a;

(2) Mix product a with hydroxylamine hydrochloride and potassium carbonate at a molar ratio of 1:6:2, dissolve in absolute ethanol, and react at 90°C for 2 hours. Then evaporate the solvent to dryness, extract with water and ethyl acetate, keep the organic layer, dry over anhydrous sodium sulfate and evaporate to dryness to obtain product b;

(3) React the product b with an equimolar amount of o-chlorobenzoic acid, the reaction solvent is dichloromethane, react at room temperature for 6 hours, evaporate the solvent to dryness after the reaction is complete, extract with water and ethyl acetate, keep the organic solvent layer, anhydrous After drying over sodium sulfate, evaporate to dryness to obtain nitroimidazole derivatives. The reaction solvent is dichloromethane, the catalyst is HOBT, and the binder is EDC.HCl, wherein the molar ratio of product b, HOBT and EDC.HCl is 1:1:2.

The product is a colorless powder, the yield is 70%, mp180.4-180.5℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.249 (s, 3H), 5.896 (s, 2H), 7.600-7.621 ( m, 2H), 7.697-7.718(m, 2H), 7.790-7.812(m, 1H), 8.410(s, 1H), 8.539-8.566(m, 1H), 9.162-9.169(m, 1H).

Example 3

When R1=R2=R4=R5=H, when R3 is Br, the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 65%, mp170.7℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.210-2.251 (m, 3H), 5.781 (s, 2H), 7.558-7.635 ( m, 4H), 7.688-7.717 (m, 2H), 7.822-7.853 (m, 1H), 7.905-7.936 (m, 1H), 8.289 (s, 1H).

Example 4

When R1=R2=R4=R5=H, and R3 is CH3, the preparation method is as in Example 1.

The product is a colorless powder, the yield is 60%, mp171.0-171.5℃; ¹ H NMR (400MHz,DMSO)δ(ppm):2.222-2.245(m,3H),2.506-2.569(m,3H), 5.763(s,2H),7.383-7.439(m,2H),7.567-7.613(m,3H),7.692-7.713(m,2H),7.985-8.003(d,J=7.2Hz,1H),8.359( s, 1H).

Example 5

When R1=R2=R4=R5=H, and R3 is _OCH3 , the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 60%, mp156.9-157.2℃; ¹ H NMR (400MHz,DMSO)δ(ppm):2.340(s,3H),2.502-2.511(m,3H),5.417( s, 2H), 7.557-7.636 (m, 5H), 7.693-7.714 (m, 1H), 7.895-7.898 (m, 2H), 8.194 (s, 1H).

Example 6

When R1=R2=R4=R5=H, and R3 is NO ₂ , the preparation method is the same as in Example 1.

The product is light yellow powder with a yield of 60%, m.p.158.5-158.6°C.

Example 7

When R1=R2=R4=R5=R3=H, the preparation method is the same as in Example 1.

The product is a colorless powder with a yield of 65%, mp183.5-183.6℃; ¹ H NMR (400MHz,DMSO)δ(ppm):2.253(s,3H),3.358(s,3H),5.866(s, 2H), 7.595-7.630(m, 4H), 7.691-7.713(m, 2H), 8.410(s, 1H).

Example 8

When R1=R2=R3=R5=H, and R4 is F, the preparation method is the same as in Example 1.

The product is a colorless powder with a yield of 60%, m.p.171.7-172.1°C.

Example 9

When R1=R2=R3=R5=H, when R4 is Cl, the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 65%, mp170.7-170.8℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.247 (s, 3H), 5.871 (s, 2H), 7.589-7.641 ( m, 3H), 7.667-7.710 (m, 2H), 7.819-7.849 (m, 1H), 8.093-8.132 (m, 2H), 8.381 (s, 1H).

Example 10

When R1=R2=R3=R5=H, when R4 is Br, the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 60%, mp172.7-172.8℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.502-2.511 (m, 3H), 5.885 (s, 2H), 7.559- 7.612(m,3H),7.690-7.712(m,2H),7.967-7.987(d,J=8.0Hz,1H),8.152-8.272(d,J=8.0Hz,1H),8.264-8.272(m, 1H), 8.412(s, 1H).

Example 11

When R1=R2=R3=R5=H, and R4 is _CH3 , the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 65%, mp159.5-159.6℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.260 (s, 3H), 3.330-3.353 (m, 3H), 5.896 ( s,2H),7.612-7.634(m,2H),7.679-7.726(m,2H),8.078-8.098(d,J=8.0Hz,3H),8.258-8.280(d,J=8.8Hz,1H) ,8.407-8.423(m,1H).

Example 12

When R1=R2=R3=R4=H, and R5 is F, the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 65%, mp171.4-172.2℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.502-2.510 (m, 3H), 5.417 (s, 2H), 7.101- 7.259 (m, 1H), 7.558-7.649 (m, 5H), 7.690-7.712 (m, 1H), 8.194 (s, 1H), 12.317 (s, 1H).

Example 13

When R1=R2=R3=R4=H, when R5 is Cl, the preparation method is the same as in Example 1.

The product is a colorless powder with a yield of 60%, m.p.170.4°C.

Example 14

When R1=R2=R3=R4=H, when R5 is Br, the preparation method is the same as in Example 1.

The product is a colorless powder with a yield of 65%, m.p.171.0-171.3°C.

Example 15

When R1=R2=R3=R4=H, and R5 is _CH3 , the preparation method is the same as in Example 1.

The product is light yellow powder, the yield is 65%, mp171.4-171.5℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.502 (s, 3H), 3.578 (s, 3H), 5.163 (s, 2H),7.146-7.243(m,3H),7.276(s,1H),7.314-7.340(d,J=10.4Hz,1H),7.419(s,1H),7.540(s,2H),7.876-7.903 (d, J=10.8Hz, 1H).

Example 16

When R1=R2=R3=R4=H, and R5 is _OCH3 , the preparation method is the same as in Example 1.

The product is a colorless powder, the yield is 60%, mp172.0-172.7℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.249-2.412 (m, 3H), 3.814-3.880 (m, 3H), 5.824 (s, 2H), 7.017-7.124 (m, 2H), 7.590-7.797 (m, 4H), 8.088-8.117 (d, J=11.6Hz, 2H), 8.302-8.357 (m, 1H).

Example 17

When R1=R2=R3=R4=H, and R5 is NO2, the preparation method is the same as in Example ₁ .

The product is light yellow powder, the yield is 65%, mp171.1-171.6℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.502-2.511 (m, 3H), 5.844-5.871 (m, 2H), 7.431-7.711 (m, 2H), 7.678-7.683 (m, 3H), 7.903-7.921 (m, 1H), 8.231-8.274 (m, 2H), 8.410 (s, 1H).

Example 18

When R1=N, R1=R4=R5=H, and R3 is Cl, the preparation method is the same as in Example 2.

The product is light yellow powder with a yield of 70%, m.p.185.5-185.6°C.

Example 19

When R1=N, R2=R3=R4=H, and R5 is _CH3 , the preparation method is the same as in Example 2.

The product is light yellow powder, the yield is 70%, mp180.9-181.0℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.245-2.251 (m, 3H), 3.366 (s, 3H), 5.417 ( s,1H),5.882(s,1H),7.502-7.621(m,4H),7.691-7.712(m,1H),8.195(s,1H),8.385-8.411(m,1H),12.318(s, 1H).

Example 20

When R1=N, R2=R3=R4=H, and R5 is _OCH3 , the preparation method is the same as in Example 2.

The product is a colorless powder, the yield is 70%, mp176.7-176.9℃; ¹ H NMR (400MHz, DMSO) δ (ppm): 2.244-2.251 (m, 3H), 3.359-3.473 (m, 3H), 5.417(s,1H),5.875(s,1H),7.014-7.013(m,1H),7.579-7.616(m,3H),7.685-7.707(m,2H),8.365-8.399(m,1H), 9.025-9.030 (m, 1H).

Example 21

When R2=N, R1=R3=R4=R5=H, the preparation method is the same as in Example 2.

The product is a light yellow powder with a yield of 65%, m.p.166.3-166.6°C.

The following anticancer activity studies were carried out on the products obtained in Examples 1-21.

1. Experimental materials

1.1 Cell lines

Human breast cancer cell line (Human breast carcinoma cells), MCF-7.

1.2 Reagents

RPMI1640 medium (GIBCO), newborn bovine serum (Hangzhou Sijiqing Bioengineering Materials), trypsin (Sigma), MTT (Sigma), streptomycin for injection (Shandong Ruiyang Pharmaceutical, 1g 1 million units/bottle), for injection Penicillin sodium (Shandong Lukang Pharmaceutical, 800,000 units/bottle). Other commonly used chemical reagents were domestic analytical grade.

2. Experimental method

2.1 Preparation of medium

Add one liter of water to one bag of RPMI1640 medium (Gibcio USA), add 2g of sodium bicarbonate, 100,000 units of penicillin and 100mg of streptomycin, adjust the pH value to 7.4, and filter and sterilize with a 0.22mm sterile filter membrane. 90mL culture medium plus 10mL inactivated newborn bovine serum is the complete culture medium. Trypsin was made into a 0.25% solution with D-hanks buffer, and stored at 4°C after filter sterilization.

2.2 Preparation of liquid medicine

Accurately weigh 1.0 mg of the sample to be tested, add it to a sterilized 1.5 mL centrifuge tube, add 1 mL of DMSO to make a 1 mg/mL stock solution, and store it in a freezer at -40 °C. Dilute with appropriate amount of D-hanks to the corresponding concentration after melting before use.

2.3 Cell culture and passage

All the cells were cultured adherently in cell culture flasks containing 10ml of complete culture medium at 37°C, 5% CO ₂ , and saturated humidity. After the cells cover the bottom of the bottle, wash twice with sterilized D-hanks solution, add 0.25% trypsin to digest the cells for 2 minutes, pour off the trypsin, and after the cells can be completely detached by shaking gently, add 30ml of complete culture solution, and pipette The tube was blown to disperse the cells, and the cells were divided into 3 new cell culture flasks, and the culture was continued.

2.4 Anticancer activity test

Take a bottle of cells that have just grown into a complete monolayer, collect the cells after trypsinization, blow evenly with a pipette, take two drops of cell suspension for Trypan Blue staining, and count the number of living cells (dead cells) under a microscope. The number of cells shall not exceed 5%), adjust the number of cells to 1×10 ⁴ cells/ml with complete culture medium. Add 100 μL of cell suspension to each well of a 96-well cell culture plate, place the culture plate in a CO2 incubator for 24 hours, take out the culture plate, and add 11 μL of solutions containing different concentrations of tested samples to each well to make the final drug concentration 60, 20, 6.67, 2.22, 0.74, 0.25, and 0.082 μg/mL, respectively. Three parallel wells were set up for each concentration, and 6 wells of cells were set up as normal control wells and positive control wells. After adding the medicine, the culture plate was shaken and mixed evenly on a microplate shaker, and placed in a CO ₂ incubator to continue culturing for 24 hours. Take out the culture plate, add 25 μL of 5 mg/mL MTT solution to each well, shake and mix well, and continue to incubate for 4 h. Add 100 μL DMSO to each well and incubate for 10 min. The light absorption (OD value) of each well was measured by a microplate reader, and the measurement wavelength was 490nm. Among them, 5-Fluorouracil a was used as a positive reference. According to the OD value of each well, the inhibitory rate of the drug on the proliferation of MCF-7 cells, that is, the _IC50 value, was calculated by CalcuSyn software. The experimental results are shown in Table 1.

The product of table 1 embodiment 1-21 is to the inhibitory rate of MCF-7 cell proliferation

_IC50 (μM) Example 1 35.6±2.3 Example 2 1.8±1.8 Example 3 13.7±1.1 Example 4 21.2±0.2 Example 5 11.3±1.5 Example 6 17.3±2.0 Example 7 25.7±1.9 Example 8 18.1±2.1 Example 9 10.8±0.2 Example 10 30.7±2.5 Example 11 28.1±2.3 Example 12 20.6±0.5 Example 13 27.6±1.9 Example 14 24.9±1.8 Example 15 15.2±1.3 Example 16 31.1±2.0 Example 17 8.3±1.6 Example 18 0.9±1.3 Example 19 0.5±0.8 Example 20 0.78±0.2 Example 21 1.21±0.3 5-Fluorouracil ^a 21.9±1.4

Claims (8)

1. a nitroimidazole derivative is characterized in that its molecular structural formula is as follows: ; in: When R ₁ =CH, R ₂ =C, R ₄ =R ₅ =H, R ₃ is Cl. 2. a preparation method of the nitroimidazole derivative as claimed in claim 1, is characterized in that, comprises the steps: (1) Dissolve 2,4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, and tetrabutylammonium bromide in absolute ethanol, react at 85°C for 10 minutes, and then wait The solution is filtered after cooling, and the filtered solid is washed twice with distilled water, filtered, and recrystallized to obtain product a, which is ; (2) Mix product a, hydroxylamine hydrochloride and potassium carbonate, dissolve in absolute ethanol, react at 90°C for 2 to 4 hours, then evaporate the solvent to dryness, extract with water and ethyl acetate, keep the organic layer, anhydrous sodium sulfate Evaporate to dryness after drying to obtain product b, product b is ; (3) React the product b with o-chlorobenzoic acid, react at room temperature for 6-12 hours, evaporate the solvent to dryness after the reaction is complete, extract with water and ethyl acetate, keep the organic solvent layer, dry it with anhydrous sodium sulfate, and evaporate to dryness to obtain nitro Imidazole derivatives; The synthetic route is as follows: . 3. the preparation method of nitroimidazole derivative according to claim 2 is characterized in that, 2,4'-dibromoacetophenone, 2-methyl-4-nitroimidazole, potassium carbonate, tetrabutyl The molar ratio of ammonium bromide is 1:1:1-3:0.05-1. 4. the preparation method of nitroimidazole derivative according to claim 2 is characterized in that, the mol ratio of product a, hydroxylamine hydrochloride and salt of wormwood is 1:1-10:1-3. 5. the preparation method of nitroimidazole derivative according to claim 2, is characterized in that, product b and o-chlorobenzoic acid mol ratio are 1:1-1.5. 6. The preparation method of nitroimidazole derivatives according to claim 2, characterized in that, in step (3), the reaction solvent is dichloromethane, and the catalyst is 6-chloro-1-hydroxybenzotriazole, The binder is EDC.HCl. 7. the preparation method of nitroimidazole derivative according to claim 6 is characterized in that, the mol ratio of product b, 6-chloro-1-hydroxyl benzotriazole and EDC.HCl is 1: 0.05-1 :1-2. 8. A use of the nitroimidazole derivatives according to claim 1, characterized in that the nitroimidazole derivatives are used in the preparation of anti-breast cancer drugs.