JP3258776B2 - N-styrylphenylaminobenzoic acid derivatives - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a compound of the general formula (1) having an inhibitory effect on tumor growth and useful as an antitumor agent.

Embedded image (Wherein, R ₁ represents a hydrogen atom or a halogen atom, R ₂
Represents a hydrogen atom or a lower alkoxy group having 1 to 3 carbon atoms, R ₃ and R ₄ are the same or different and each represent a hydrogen atom, a lower alkyl group having 1 to 3 carbon atoms, or a halogen atom;
Represents a nitrogen atom or CH), which is an N-styrylphenylaminobenzoic acid derivative represented by the formula:

[0002]

2. Description of the Related Art Many of the currently used antitumor agents attack cancer cells to exert a certain antitumor effect, but also exhibit toxicity to normal cells.
Due to the problem of exhausting the patient's physical strength, it is difficult to use it for a long period of time, and often cannot bring complete healing of the tumor. Therefore, development of a new antitumor agent that exhibits an antitumor effect without showing toxicity to normal cells has been awaited today.

[0003]

Means for Solving the Problems The present inventors have been interested in the physiology of cancer cells and their malignant forms and have been conducting research on synthesizing various compounds to normalize the malignant forms of cancer cells. As a result, some N-styrylphenylaminobenzoic acid derivatives and N-styrylphenyl-2-aminonicotinic acid derivatives strongly suppress the growth of cancer cells, but do not affect the growth of normal cells. Was found. The present invention has been completed by further research based on this finding. That is, the present invention relates to a compound represented by the general formula (1).

In the compound of the present invention, the lower alkyl group having 1 to 3 carbon atoms is specifically a methyl group, an ethyl group or a propyl group, and the halogen atom is a fluorine atom, a chlorine atom or the like. Examples of the lower alkoxy groups 1 to 3 include a methoxy group and an ethoxy group.

The compound represented by the general formula (1) of the present invention is a novel compound and can be produced, for example, by the method shown below.

Embedded image

Embedded image

Embedded image (Wherein R1, R2, R3, and R4 are the same as described above, and Y represents a bromine atom or an iodine atom.)

The starting material diphenyliodonium-2-carboxylate (compound 2) in the above reaction formula
Are described in the literature, for example, Robert A. et al. Sche
rrer et al. (J. Org. Chem. 45, 21).
27-2131, 1980). Compound 3 is obtained by a Wittig reaction between a p-nitrobenzaldehyde derivative and a benzyltriphenylphosphonium bromide derivative to obtain a trans-4-nitrostilbene derivative, which is reduced with iron and hydrochloric acid to give the desired compound 3. can get.

Hereinafter, the present invention will be described specifically with reference to examples, but the present invention is not limited to these examples.

[0008]

【Example】

Example 1 4- (2'-Methylstyryl) -aniline (0.02 mol) was dissolved in dry DMF, and diphenyliodonium-2-carboxylate was dissolved in 0.02 mol of the solution.
A mole and 150 mg of ferric acetate are added and heated at 85-90 ° C. for 12 hours with stirring under a nitrogen stream. Water is added to the reaction solution, and DMF is concentrated under reduced pressure, made acidic with dilute hydrochloric acid and extracted with ethyl acetate. After washing the organic layer with saturated saline,
After drying over magnesium sulfate, filtration and concentration of the solvent, crystals are obtained. When the crystals are recrystallized with a benzene-methanol mixed solvent, 4 '-(2 "-methylstyryl) -diphenylamine-2-carboxylic acid (compound a) is obtained in a yield of 80%.

Similarly, 4 '-(2 "-ethylstyryl) -diphenylamine-2-carboxylic acid (compound b)
And 2'-methoxy-4 '-(2 "-methylstyryl) -diphenylamine-2-carboxylic acid (compound c)
In a yield of 75% and 60%, respectively.

[0010]

Example 2 2-Iodo-4-fluorobenzoic acid (0.0
1 mol) and 4- (2'-methylstyryl) -2-methoxyaniline (0.0113 mol) in styrene (15 m
l), 100 mg of copper powder and 2.1
g of t-Bu0K is added and refluxed for 1 hour with stirring under a nitrogen stream. The reaction solution is poured into dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate layer is washed with saturated saline, dried over magnesium sulfate, filtered, and the solvent is concentrated under reduced pressure to obtain an oily substance. This oily substance was purified by silica gel column chromatography (eluted with toluene) to give 4 '-(2 "-methylstyryl) -2'-methoxy-4-fluorodiphenylamine-2-carboxylic acid (compound d) in a yield of 20%. Get)

[0011]

Example 3 2-chloronicotinic acid (0.02 mol) and 4- (2'-methylstyryl) -aniline were dissolved in 2-methoxyethanol, and the mixture was refluxed for 5 hours under a nitrogen stream. The reaction solution is poured into water containing 0.02 mol of sodium hydrogen carbonate and extracted with ethyl acetate. The ethyl acetate layer is washed with a saturated saline solution and dried with magnesium sulfate. After filtration, the solvent is concentrated under reduced pressure to obtain crystals. The crystals were recrystallized with a mixed solvent of benzene-methanol to give N-4 '-(2 "-methylstyryl) phenyl-2 in a yield of 50%.
-Obtaining aminonicotinic acid (compound e).

Similarly, N-4 '-(2 "-chlorostyryl) phenyl-2-aminonicotinic acid (compound f),
N-4 '-(2 ", 6" -dichlorostyryl) phenyl-2-aminonicotinic acid (compound g), N-4'-
(2 ", 6" -difluorostyryl) phenyl-2-aminonicotinic acid (Compound h), N-4 '-(2 ", 6"-
Dimethylstyryl) phenyl-2-aminonicotinic acid (Compound i) was obtained in yields of 40, 50, 60 and 55%, respectively.

The physical properties of the compounds a to i obtained in Examples 1 to 3 are shown in Table 1.

[Table 1]

[0014]

[Experimental example 1] 10% (volume%) fetal calf serum (hereinafter abbreviated as FCS)
Dulbecco Modified Eagle Medium (hereinafter Dulbecco MEM. Gibco B. Inc.)
Human fibrosarcoma cell line HT-10 grown semi-confluently in R.L. (GIBCO BRL)
After trypsinization, 80 was suspended at a cell concentration of 2.5 × 10 ⁴ cells / ml using Dulbecco MEM containing 10% FCS, and the suspension was dispensed into a 24-well multiwell dish, 1 ml per well. . Then 37 ° C, 5
After culturing for 24 hours under% CO ₂ , the medium was removed and 1 ml of Dulbecco's MEM containing 10% FCS containing serially diluted test compound was added. Only the solvent in which the test compound was dissolved was added to the control wells. After addition of test compound, the cells are then cultured for 96 hours Ca ² + and Mg
After washing with ^{2 +} free phosphate buffer, 50 per well
The cells were recovered as a suspension by adding 0 μl of 0.25% trypsin / 0.02% sodium ethylenediaminetetraacetate solution and incubating at 37 ° C. for 10 minutes. One part of the cells was collected in 5 ml of Isoton III (isotonic. In a vial containing a hemocytometer diluent (manufactured by Nikkaki Co., Ltd.), and the number of cells was measured with a Coulter counter. n =
From the results obtained in step 2, the concentration of each compound that suppresses cancer cell proliferation by 50% was determined. Table 2 shows the results.

[Table 2]

[0015]

[Experimental example 2] Implantation subcutaneously into C57BL / 6 mice
4 days mice B16 melanoma grown aseptically excised, 1 group 8 mice BDF ₁ mice after the free cells by Dispase treatment (6 weeks old, male) were flank subcutaneously ¹⁰⁶ transplantation. 24 hours after transplantation, compounds e and f were orally administered at 20, 40, 60 mg / kg body weight once a day.
The administration was continued until the first day, the tumor was excised on the twelfth day, the weight was measured, and the tumor growth inhibition rates of the compound e and f administration groups with respect to the control group were determined by the following formula and compared. Table 3 shows the results. Growth inhibition rate (%) = (CT) / C × 100 T: average tumor weight of administration group (mg) C: average tumor weight of control group (mg)

[Table 3]

Continuation of the front page (58) Field surveyed (Int. Cl. ⁷ , DB name) C07C 229/58 C07D 213/80 CA (STN) REGISTRY (STN)

Claims (1)

(57) [Claims] 1. A compound of the general formula (Wherein, R ₁ represents a hydrogen atom or a halogen atom, R ₂
Represents a hydrogen atom or a lower alkoxy group having 1 to 3 carbon atoms, R ₃ and R ₄ are the same or different and each represent a hydrogen atom, a lower alkyl group having 1 to 3 carbon atoms, or a halogen atom;
Represents a nitrogen atom or CH). N-styrylphenylaminobenzoic acid derivative represented by the formula: