CN107556309B

CN107556309B - Pharmaceutical application of polysubstituted tetrahydronaphthyridine compound and preparation method thereof

Info

Publication number: CN107556309B
Application number: CN201710814057.9A
Authority: CN
Inventors: 叶天健; 陈鑫; 叶继华; 王晨竹; 蔡翔
Original assignee: Zhejiang Yongning Pharmaceutical Co Ltd
Current assignee: Zhejiang Yongning Pharmaceutical Co Ltd
Priority date: 2017-09-11
Filing date: 2017-09-11
Publication date: 2020-12-01
Anticipated expiration: 2037-09-11
Also published as: CN107556309A

Abstract

The invention provides a pharmaceutical application and a preparation method of a polysubstituted tetrahydronaphthyridine compound with the structural formula as follows.

Description

Pharmaceutical application of polysubstituted tetrahydronaphthyridine compound and preparation method thereof

Technical Field

The invention belongs to compound application, relates to medicinal application and a preparation method of a novel polysubstituted tetrahydronaphthyridine compound, and particularly relates to application of the polysubstituted tetrahydronaphthyridine compound in the aspect of tumor resistance and a preparation method of the polysubstituted tetrahydronaphthyridine compound.

Background

Malignant tumors are one of the major diseases that currently seriously affect human health and threaten human life. Cancer, together with cardiovascular and cerebrovascular diseases and accidents, constitutes three leading causes of death in all countries of the world today. Therefore, the world health organization and the government health departments of all countries have cancer as a first task to overcome. Therefore, the development of the anti-tumor drugs has extremely important social significance and application value.

The naphthyridine compounds have wide biological activity and pharmacological activity. Meanwhile, the naphthyridine compounds are important intermediates for preparing other heterocyclic compounds. Therefore, the research on the application of the polysubstituted tetrahydronaphthyridine compound in the aspect of tumor resistance has important significance.

Disclosure of Invention

The invention aims to provide application of a novel polysubstituted tetrahydronaphthyridine compound in preparing an anti-tumor medicament, in particular application in resisting lung cancer caused by a cell strain A549 and leukemia caused by a cell strain HL-60. The general formula of the polysubstituted tetrahydronaphthyridine compound is as follows:

wherein R is phenyl, 4-methylphenyl, 4-fluorophenyl or 4-chlorphenyl, and is characterized in that the polysubstituted tetrahydronaphthyridine compound is applied to resisting lung cancer and leukemia.

The polysubstituted tetrahydronaphthyridine compound is a compound I-1, I-2, I-3 or I-4, and is shown as follows:

wherein the compounds I-1, I-3 and I-4 are applicable to the application of resisting the lung cancer caused by the cell strain A549. The compound I-2 is applicable to resisting leukemia caused by a cell strain HL-60.

The novel polysubstituted tetrahydronaphthyridine compound is synthesized by taking N-methyl-piperidine-2-ketone, R-CHO, ammonium acetate and malononitrile as raw materials through a one-pot condensation reaction. The synthetic route is as follows:

the novel polysubstituted tetrahydronaphthyridine compound provided by the invention has good antitumor activity, has strong inhibition effect on tumor cell strains A549 and HL-60, lays a foundation for new drug screening and development, has the advantages of low price, easily available raw materials, suitability for industrial production and the like, and has good practical value.

Detailed Description

The present invention is further illustrated by the following examples, which are not intended to limit the scope of the invention.

The methods used in the following examples are conventional methods unless otherwise specified. Materials or reagents required in the following examples are commercially available unless otherwise specified.

Example 1

Preparation of Compound I-1:

113g (1mol) of N-methyl-piperidine-2-ketone, 212g (2mol) of benzaldehyde, 77g (1mol) of ammonium acetate, 66g (1mol) of malononitrile and 15g of trifluoromethanesulfonic acid are sequentially added into a single-mouth bottle, and the temperature is raised to 100 ℃ for reaction for 5 hours. After the reaction is finished, cooling to room temperature, sequentially adding 1000mL of ethyl acetate and 500mL of water, taking an organic layer, adding 500mL of saturated sodium bicarbonate solution for washing, taking the organic layer, drying with anhydrous sodium sulfate, filtering, concentrating to 200mL, stirring at room temperature for crystallization to obtain a compound I-1303 g with the yield of 86%.¹H NMR:7.40(m,11H),5.23(brs,2H),3.21(s,3H),3.15(t,J＝7.0Hz,2H),2.13(t,J＝7.0Hz,2H).

Example 2

Preparation of Compound I-2:

113g (1mol) of N-methyl-piperidin-2-one, 240g (2mol) of p-tolualdehyde, 77g (1mol) of ammonium acetate, 66g (1mol) of malononitrile and 15g of trifluoromethanesulfonic acid were sequentially added to a single-neck flask, and the mixture was heated to 100 ℃ to react for 5 hours. After the reaction is finished, cooling to room temperature, sequentially adding 1000mL of ethyl acetate and 500mL of water, taking an organic layer, adding 500mL of saturated sodium bicarbonate solution for washing, taking the organic layer, drying with anhydrous sodium sulfate, filtering, concentrating to 200mL, stirring at room temperature for crystallization to obtain compound I-2274 g, wherein the yield is 72%.¹H NMR:7.42(m,9H),5.01(brs,2H),3.21(s,3H),3.15(t,J＝7.0Hz,2H),2.34(s,6H),2.13(t,J＝7.0Hz,2H).

Example 3

Preparation of Compound I-3:

113g (1mol) of N-methyl-piperidin-2-one, 248g (2mol) of p-fluorobenzaldehyde, 77g (1mol) of ammonium acetate and 66g (1mol) of malononitrile were sequentially added to a single-neck flaskAnd 15g of trifluoromethanesulfonic acid, and heating to 100 ℃ to react for 5 hours. After the reaction is finished, cooling to room temperature, sequentially adding 1000mL of ethyl acetate and 500mL of water, taking an organic layer, adding 500mL of saturated sodium bicarbonate solution for washing, taking the organic layer, drying with anhydrous sodium sulfate, filtering, concentrating to 200mL, stirring at room temperature for crystallization to obtain a compound I-3291 g with the yield of 75%.¹H NMR:7.47(m,9H),5.16(brs,2H),3.21(s,3H),3.12(t,J＝7.0Hz,2H),2.11(t,J＝7.0Hz,2H).

Example 4

Preparation of Compound I-4:

113g (1mol) of N-methyl-piperidine-2-ketone, 280g (2mol) of p-chlorobenzaldehyde, 77g (1mol) of ammonium acetate, 66g (1mol) of malononitrile and 15g of trifluoromethanesulfonic acid are added into a single-mouth bottle in sequence, and the temperature is raised to 100 ℃ for reaction for 5 hours. After the reaction is finished, cooling to room temperature, sequentially adding 1000mL of ethyl acetate and 500mL of water, taking an organic layer, adding 500mL of saturated sodium bicarbonate solution for washing, taking the organic layer, drying with anhydrous sodium sulfate, filtering, concentrating to 200mL, stirring at room temperature for crystallization to obtain compound I-4307 g, wherein the yield is 73%.¹H NMR:7.43(m,9H),5.11(brs,2H),3.20(s,3H),3.10(t,J＝7.0Hz,2H),2.05(t,J＝7.0Hz,2H).

Example 5

anti-A549 tumor cell bioactivity test

In vitro anti-tumor activity test method:

1. sample preparation: compounds I-1, I-2, I-3 and I-4 obtained in examples 1 to 4 were dissolved in 20. mu.L of DMSO per 1mg, and 2. mu.L of the solution was diluted with 1000. mu.L of the culture solution to a concentration of 100. mu.g/mL, followed by serial dilution of the culture solution to the use concentration.

2. Culture of cells

Preparation of a culture medium: every 1000mL of culture medium contains 80 million units of penicillin, 1.0g of streptomycin and 10% inactivated fetal calf serum;

and (3) culturing the cells: a549 tumor cells were inoculated in a medium and incubated at 37 ℃ with 5% CO₂Culturing in an incubator for 5 days for passage.

3. Determination of the inhibition of tumor cell growth by samples

Digesting the cells with EDTA-trypsin digest and diluting with mediumReleased into 1 × 10⁵Perml, 100. mu.L/well in 96-well cell culture plates, 37 ℃ 5% CO₂Culturing in an incubator. After 24h of inoculation, samples diluted with medium were added, 100. mu.L per well, 3 wells per concentration, and placed at 37 ℃ in 5% CO₂Culturing in an incubator. After 72h, adding 5mg/mL MTT into a cell culture well, keeping the cell culture well at 10 mu L per well, incubating the cell culture well at 37 ℃ for 4h, adding DMSO into the cell culture well at 150 mu L per well, uniformly oscillating, carrying out color comparison by using an enzyme-linked immunosorbent assay (ELISA) instrument at a wavelength of 570nm, using a blank control without a sample and cells cultured by a culture medium containing DMSO with the same concentration as a control under the same conditions, and calculating the half lethal concentration (IC) of the sample on the growth of tumor cells₅₀)。

In addition, the test method for in vitro antitumor activity was performed by the above method, using carboplatin as a control. The results are shown in table 1:

TABLE 1 IC of Compounds on A549 tumor cells₅₀(μg/mL)

Examples	Compound (I)	IC₅₀
			1	I-1	7.21
2	I-2	>100
			3	I-3	9.11
4	I-4	5.25
			Control drug	Carboplatin	5.11

The results show that the compounds I-1, I-3 and I-4 have obvious inhibitory activity on A549 tumor cells.

Example 6

anti-HL-60 tumor cell bioactivity test

In vitro anti-tumor activity test method:

4. sample preparation: compounds I-1, I-2, I-3 and I-4 obtained in example 1-4 were dissolved in 20. mu.L of LDMSO per 1mg, 2. mu.L of the solution was diluted with 1000. mu.L of the culture medium to a concentration of 100. mu.g/mL, and the solution was then serially diluted with the culture medium to the use concentration.

5. Culture of cells

and (3) culturing the cells: inoculating HL-60 tumor cells into culture medium, standing at 37 deg.C and 5% CO₂Culturing in an incubator for 5 days for passage.

6. Determination of the inhibition of tumor cell growth by samples

Cells were digested with EDTA-trypsin digest and diluted to 1X 10 with medium⁵Perml, 100. mu.L/well in 96-well cell culture plates, 37 ℃ 5% CO₂Culturing in an incubator. After 24h of inoculation, samples diluted with medium were added, 100. mu.L per well, 3 wells per concentration, and placed at 37 ℃ in 5% CO₂Culturing in an incubator. Adding 5mg/mL MTT into the cell culture well after 72h, incubating at 37 deg.C for 4h with 10 μ L MTT per well, adding DMSO with 150 μ L DMSO per well, shaking uniformly, and measuring with microplate reader at 570nm wavelengthColorimetric determination of the median lethal concentration (IC) of a sample on tumor cell growth was calculated using the same conditions with a blank control without the sample and cells cultured in medium containing the same concentration of DMSO as a control₅₀)。

In addition, the drug cytarabine is selected as a control, and the in vitro anti-tumor activity test method is carried out according to the method. The results are shown in table 2:

TABLE 2 IC of Compounds on HL-60 tumor cells₅₀(μg/mL)

Examples	Compound (I)	IC₅₀
			1	I-1	>100
2	I-2	2.59
			3	I-3	>50
4	I-4	>50
			Control drug	Cytarabine	3.59

The results show that the compound I-2 has obvious inhibition effect on HL-60 tumor cells.

After reading the description of the method, one skilled in the art can make various changes or modifications to the invention, and such equivalents fall within the scope of the claims appended to the present application.

Claims

1. The application of polysubstituted tetrahydronaphthyridine compounds in preparing medicaments for treating leukemia tumors is disclosed, wherein the polysubstituted tetrahydronaphthyridine compounds are compounds I-2 shown in the following figures:

2. the application of polysubstituted tetrahydronaphthyridine compounds in medicaments for treating lung cancer tumors is shown in the following figures, wherein the polysubstituted tetrahydronaphthyridine compounds are compounds I-1, I-3 and I-4:

3. a process for the preparation of polysubstituted tetrahydronaphthyridines according to claim 1 or 2, characterized in that: the N-methyl-piperidine-2-ketone, R-CHO, ammonium acetate and malononitrile are taken as raw materials and synthesized by a one-pot condensation reaction, and the synthetic route is as follows:

wherein R is phenyl, 4-methylphenyl, 4-fluorophenyl or 4-chlorophenyl.