CN105418608B - 7 benzos [b] [1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide and its production and use - Google Patents

Abstract

The invention discloses a kind of 7 benzos [b] [1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide and its production and use, its structural formula is

Description

7-Benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea and preparation method thereof and use

technical field

The present invention relates to the technical field of medicine, in particular to the technical field of antitumor drugs, in particular to a preparation method of 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea with antitumor activity and use

Background technique

Acridine is a class of nitrogen-containing organic heterocyclic compounds that has received widespread attention. Because its structure is a macrocyclic conjugated system with a rigid planar structure, it can be used as an intercalator for macromolecules such as DNA. It has anti-tumor, anti-virus, and anti-malarial , antibacterial, bioluminescent probes and the treatment of AIDS, etc. have shown strong physiological activities. People have obtained a large number of acridine compounds from natural products or by chemical synthesis. Their pharmacological activities and mechanism of action. In order to improve its affinity to DNA, the inhibitory performance of various enzymes and cytotoxicity, etc., its structural transformation has been continuously explored and improved. The innovation of the present invention lies in the use of benzo[b]-[1 , 10] The 7-position of the o-phenanthroline ring is connected with an active group amidothiourea structure to synthesize a novel acridine derivative, which has not yet been reported.

Contents of the invention

The purpose of the present invention is to overcome the above problems and provide a compound with anti-tumor activity, which is connected with an active group amidothiourea at the 7-position of the benzo[b]-[1,10]phenanthroline ring Structure, synthesis of novel acridine derivatives, which have strong anti-tumor activity and can be used in the preparation of anti-tumor drugs.

Another object of the present invention is to provide a method for preparing a drug with anti-tumor activity, and its structural modification improves the affinity of the anti-tumor drug to DNA, the consistency of various enzymes, and the cytotoxicity.

Another object of the present invention is to provide a kind of 7-benzo [b]-[1,10] o-phenanthroline pyridine carboxamide thiourea in the preparation of antitumor drug application.

In order to realize the purpose and other advantages of the present invention, a kind of 7-benzo [b]-[1,10] o-phenanthroline pyridine carboxamide thiourea is provided, and its structural formula is:

The 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea has a molecular formula of C ₂₃ H ₁₆ N ₆ OS, a relative molecular weight of 424.48, and physical and chemical properties: yellow powder, mp175-181 ℃; ¹ H NMR (DMSO-d ₆ , 400M Hz), δ: 12.89 (br, s, 1H, -NH), 12.41 (br, s, 1H, -NH), 11.07 (br, s, 1H, -NH) NH), 10.99(s, 1H, ArH), 10.22(d, 1H, J=9.0, ArH), 9.09(d, 1H, J=8.5, ArH), 8.82(s, 1H, ArH), 8.64(d , 2H, J=7.2, ArH), 8.47(d, 1H, J=7.2, ArH), 8.24-8.36(m, 2H, ArH), 7.86-7.96(m, 2H, ArH), 7.83(d, 1H , J=7.2, ArH), 7.77 (d, 1H, J=7.2, ArH), 7.66-7.74 (m, 2H, ArH); ¹³ C NMR (DMSO-d ⁶ , 100MHz), δ189.70, 170.21, 150.34, 149.89, 145.21, 134.38, 133.78, 133.42, 132.18, 131.29, 130.55, 130.21, 129.67, 129.27, 128.54, 127.64, 126.79, 124.35, 123.78.

The preparation method of 7-benzo[b]-[1,10]o-phenanthroline pyridine carboxamide thiourea is prepared by the following synthetic route:

Preferably, the preparation method of the 7-benzo[b]-[1,10]-phenanthroline pyridinecarboxamide thiourea, the specific steps are:

Step 1, add 20-30 parts of o-bromobenzoic acid and 30-40 parts of 8-aminoquinoline in the container according to weight parts as raw materials, 6-8 parts of potassium carbonate and 0.2-0.4 parts of copper powder as catalysts, add 20- 50mL of n-amyl alcohol or isoamyl alcohol is used as a solvent, heated to 130-150°C and refluxed for 1.5-3 hours. After the reaction is completed, the solvent is evaporated under reduced pressure, and then water is added to the solid residue to continue the reaction for 15-30 minutes. Control the temperature at 70-90°C, filter, adjust the pH of the filtrate, and filter with suction to obtain the compound N-quinolyl anthranilic acid;

Step 2, mix the obtained compound N-quinolyl anthranilic acid with 7-7.5mL pure phosphorus oxychloride, heat in an oil bath, and react for 1-3 hours to obtain the compound 7-chlorobenzo[b]- [1,10] o-phenanthroline;

Step 3, add 0.2-0.4 parts of the compound 7-chlorobenzo[b]-[1,10]phenanthroline and 25-35mL acetone into the container, add 0.275 parts of sodium thiocyanate and 0.0075 Tetrabutylammonium bromide, continue to reflux reaction for 2-4 hours, until yellow solid powder precipitates, filter with suction, wash with water to obtain compound 7-benzo[b]-[1,10]phenanthroline isothiocyanate ester;

Step 4, add 0.2-0.3 parts of the compound 7-benzo[b]-[1,10]phenanthroline isothiocyanate and 20-40mL acetonitrile solution in the container, then add isoniazid, reflux reaction After 1-3 hours, a large amount of yellow powder solids were precipitated during the reaction, and the target product 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea was obtained after suction filtration and washing.

Preferably, in the preparation method of the 7-benzo[b]-[1,10]phenanthroline pyridinecarboxamide thiourea, in the step 1, the pH of the filtrate is adjusted, and concentrated hydrochloric acid is added to the filtrate to adjust pH to 1.5-2.5.

Preferably, in the preparation method of the 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea, the compound N-quinolyl anthranilic acid and three When phosphorus oxychloride is ring-closed, heat the oil bath to 85-90°C within 10-15 minutes; when a violent reaction occurs, remove the hot bath immediately; if the reaction is too violent, cool the flask with cold water until the boiling slows down and the temperature of the oil bath rises As high as 135-140°C, react for 1-3 hours.

Preferably, in the preparation method of the 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea, after the reaction in the step 2 is completed, the residue is poured into a weight ratio of In a 1:1-2:1-2 mixture of concentrated ammonia water, crushed ice, and chloroform, the solid matter was dissolved, and the chloroform layer was separated, and the aqueous layer was extracted with chloroform for 2-3 times, and the chloroform extract was combined and dried for 10-24 hours, filtered, and evaporated to remove the solvent to obtain the compound 7-chlorobenzo[b]-[1,10]phenanthroline.

Application of 7-benzo[b]-[1,10]-phenanthroline pyridine carboxamide thiourea in the preparation of antitumor drugs.

The beneficial effects of the present invention are that the present invention synthesizes novel acridine derivatives by linking the active group amidothiourea structure at the 7-position of the benzo[b]-[1,10]phenanthroline ring, improving The affinity of acridine derivatives to DNA, the inhibitory properties of various enzymes and the cytotoxicity have been studied, which provides a new idea for acridine-type anti-tumor substances.

detailed description

Example 1

Preparation of 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea

Step 1, in a 250mL three-necked flask, add 26g o-bromobenzoic acid, 34g (34mmoL) 8-aminoquinoline, 7.5g (36.2mmoL) potassium carbonate and 0.3g (4.7mmoL) copper powder, then add 30mL isoamyl Alcohol was used as a solvent, heated to 140°C and stirred at reflux for 2h. After the reaction, evaporate the solvent under reduced pressure, add 600mL of water to the obtained residue, control the temperature at 80°C for 20 minutes, filter while it is hot, wash the filter cake, combine the water layer, acidify the water layer with concentrated hydrochloric acid to pH2, and precipitate a large amount of black powder , suction filtration, and the obtained solid was recrystallized with acetone to obtain the compound N-quinolyl anthranilic acid with a yield of 36%;

Step 2, in a 100mL round bottom flask, add the obtained compound N-quinolyl anthranilic acid (9moL) and 7.2mL of pure phosphorus oxychloride, and heat the reactant to 85°C on an oil bath within 15min; When a violent reaction occurs, remove the heat bath immediately; if the reaction is too violent, cool the flask with cold water, wait until the boiling slows down, raise the temperature of the oil bath to 135°C, and react for 2 hours. After the reaction was finished, the residue was slowly poured into a well-stirred mixture of concentrated ammonia water, crushed ice and chloroform in a weight ratio of 1:1:2 according to the weight ratio after cooling, and the flask was washed with a mixture of chloroform and ammonia water. After 30 minutes, no There are undissolved solids again, separate the chloroform layer, continue to extract the water layer with chloroform for 3 times, combine the chloroform extracts, dry with anhydrous calcium chloride for 10 hours, filter, and evaporate the solvent to obtain the compound 7-chlorobenzo[ b]-[1,10]-phenanthroline, yield 18%;

Step 3: In a 100mL round bottom flask, add 0.3g (1.1mmoL) 7-chlorobenzo[b]-[1,10]phenanthroline and 30mL acetone, and add 0.275g NaSCN (3.3mmoL) after refluxing to dissolve and 0.075g (0.23mmoL) tetrabutylammonium bromide, after reflux reaction for 3h, a yellow solid powder was precipitated, filtered by suction, and washed with water to obtain compound 7-benzo[b]-[1,10]phenanthroline Isothiocyanates, 94%;

Step 4, in a 100mL round bottom flask, add 0.24g (0.8mmoL) compound 7-benzo[b]-[1,10]phenanthroline isothiocyanate and 35mL acetonitrile, then add 1mmoL benzoyl Hydrazine, reflux reaction for 2h, a large amount of solids were precipitated during the reaction, and the golden yellow solid was obtained by cooling and suction filtration, which was 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea, and the yield was 57.2 %, mp175-181°C; ¹ HNMR (DMSO-d ₆ , 400M Hz), δ: 12.89 (br, s, 1H, -NH), 12.41 (br, s, 1H, -NH), 11.07 (br, s , 1H, -NH), 10.99 (s, 1H, ArH), 10.22 (d, 1H, J=9.0, ArH), 9.09 (d, 1H, J=8.5, ArH), 8.82 (s, 1H, ArH) , 8.64 (d, 2H, J=7.2, ArH), 8.47 (d, 1H, J=7.2, ArH), 8.24-8.36 (m, 2H, ArH), 7.86-7.96 (m, 2H, ArH), 7.83 (d, 1H, J=7.2, ArH), 7.77 (d, 1H, J=7.2, ArH), 7.66-7.74 (m, 2H, ArH); ¹³ C NMR (DMSO-d ⁶ , 100MHz), δ189. 70, 170.21, 150.34, 149.89, 145.21, 134.38, 133.78, 133.42, 132.18, 131.29, 130.55, 130.21, 129.67, 129.27, 128.54, 127.64, 126.79, 124.78, 123. Made into injections, tablets, pills, capsules, suspensions or emulsions.

Example 2

In vitro anti-tumor activity experiment

1. Cell culture and passage

The selected cell lines were all placed in an incubator at 37°C and fully humidified with 5% CO2, and inoculated in PPMI1640 medium containing 10% inactivated newborn bovine serum for culture. Use an inverted microscope to observe the growth of the cells, replace the medium 2-3 times a week, passage once every 6-7 days, digest and passage with 0.25% trypsin when inoculating, usually take 3-4 passages, and use it for cells in the logarithmic growth phase in the experiment.

2. Preparation of liquid medicine

Accurately weigh the sample to be tested, add it to a sterilized 1.5mL centrifuge tube, add DMSO to make a 2mM compound stock solution, and store it in a freezer at -20°C. Dilute with appropriate amount of D-hanks to the corresponding concentration after melting before use. The concentrations of the selected compounds were determined to be 20 uM.

3. MTT experimental method

Take the cells in the logarithmic growth phase, inoculate 180uL (about 4500-5000 cells) of cell-containing medium per well on a 96-well culture plate, and culture them at 37°C and 5% _CO under fully humidified conditions for 24h. After the cells adhered to the wall, add samples in an amount of 20 uL per well, set 6 duplicate wells for each sample, and set corresponding blank controls at the same time. After continuing to culture for 48 hours, add 10uL MTT reagent (concentration: 2mg/mL) to each well. After continuing to incubate for 4h, discard the supernatant, add 150uL DMSO to each well, and shake gently for 5-8min to fully dissolve the crystal particles. The blank control group was adjusted to zero, and the absorbance value ( value), the _IC50 values of the corresponding cell lines were made with 5 concentration gradients, and all experiments were repeated 3 times to obtain the average value.

Table 1 The half effective concentration (IC ₅₀ ) of 7-benzo[c]acridinebenzamide thiourea on tumor cell lines

Table 1

As can be seen from the results of Example 2, the 7-benzo[b]-[1,10]phenanthroline pyridine carboxamide thiourea of the present invention shows through in vitro antitumor experiments that the compound has strong antitumor active. The invention provides a new idea for researching and developing new acridine type antitumor drugs.

Although the embodiment of the present invention has been disclosed as above, it is not limited to the use listed in the specification and implementation, it can be applied to various fields suitable for the present invention, and it can be easily understood by those skilled in the art Therefore, the invention is not limited to specific details without departing from the general concept defined by the claims and their equivalents.

Claims (6)

1. A kind of 1. 7- benzos [b]-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide, it is characterised in that the acridine derivatives Structural formula be：

   The 7- benzos [b]-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide Preparation method, concretely comprise the following steps：

   Step 1,20-30 parts o-bromobenzoic acid and 30-40 part 8- aminoquinolines are added in a reservoir according to parts by weight as former Material, 6-8 parts potassium carbonate and 0.2-0.4 parts copper powder be catalyst, and then addition 20-50mL n-amyl alcohols or isoamyl alcohol be as solvent, 130-150 DEG C of backflow 1.5-3 hour is heated to, after reaction terminates, solvent is removed under reduced pressure, water is then added into solid residue Continue to react 15-30 minutes, reaction temperature is controlled at 70-90 DEG C, filtering, filtrate is adjusted into pH, is filtered, is obtained compound N- quinolyl ortho-aminobenzoic acids；

   Step 2, obtained compound N-quinolyl ortho-aminobenzoic acid is mixed with the pure POCl3s of 7-7.5mL, oil bath adds Heat, 1-3 hours are reacted, compound 7- chlorobenzenes simultaneously [b]-[1,10] o-phenanthroline is made；

   Step 3,0.2-0.4 parts compound 7- chlorobenzenes simultaneously [b]-[1,10] o-phenanthroline and 25-35mL are added in a reservoir Acetone, 0.275 part of sodium sulfocynanate and 0.0075 part of TBAB are added after backflow dissolving, continues back flow reaction 2-4 hours, To there is the precipitation of yellow solid powder, filter, washing obtains compound 7- benzos [b]-[1,10] o-phenanthroline isothiocyanates；

   Step 4, add in a reservoir 0.2-0.3 parts by compound 7- benzos [b]-[1,10] o-phenanthroline isothiocyanates and 20-40mL acetonitriles, then add isoniazid, back flow reaction 1-3 hours, have a large amount of yellow powder solids to separate out in course of reaction, Target product 7- benzos [b]-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide is produced after filtering and washing.
2. 2. a kind of preparation method of 7- benzos [b]-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide as claimed in claim 1, Characterized in that, it is made by following synthetic route：
3. 3. the preparation method of 7- benzos [b] as claimed in claim 1-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide, it is special Sign is, filtrate is adjusted into pH in the step 1, and concentrated hydrochloric acid regulation pH to 1.5-2.5 is added into filtrate.
4. 4. the preparation method of 7- benzos [b] as claimed in claim 1-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide, it is special Sign is, when compound N-quinolyl ortho-aminobenzoic acid is with POCl3 cyclization in the step 2, oil bath in 10-15min It is heated to 85-90 DEG C；When vigorous reaction occurs, heating bath is removed immediately；If reaction is excessively fierce, flask can be cooled down with cold water, Treat that boiling eases up, oil bath temperature is increased to 135~140 DEG C, reacts 1-3 hours.
5. 5. the preparation method of 7- benzos [b] as claimed in claim 4-[1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide, it is special Sign is, after reaction terminates in the step 2, residue is poured into weight ratio for 1:1-2:1-2 concentrated ammonia liquor, trash ice and chlorine In imitative mixture, solids dissolving, chloroform layer is isolated, water layer continues to be extracted 2-3 times with chloroform, merges chloroform extracted solution, 10-24 hours are dried, filtering, solvent is evaporated off, that is, obtains compound 7- chlorobenzenes simultaneously [b]-[1,10] o-phenanthroline.
6. A kind of 6. 7- benzos [b]-application of [1,10] o-phenanthroline pyridinecarboxylic amido thiocarbamide in antineoplastic is prepared.