RU2707972C1 - 4-(2,4-dimethoxyphenyl)-2-(2-hydroxyphenyl)-5,6-dihydro-4n-benzo[h]chromene-3-carboxylic acid, having cytotoxic activity - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: present invention relates to 4-(2,4-dimethoxyphenyl)-2-(2-hydroxyphenyl)-5,6-dihydro-4H-benzo[h]chromene-3-carboxylic acid of said formula, having cytotoxic activity.

EFFECT: disclosed is 4-(2,4-dimethoxyphenyl)-2-(2-hydroxyphenyl)-5,6-dihydro-4n-benzo[h]chromene-3-carboxylic acid, having cytotoxic activity.

1 cl, 1 tbl, 3 dwg, 1 ex

Description

The invention relates to the field of organic chemistry, specifically to the production of 4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid having cytotoxic activity, in particular, the ability to suppress the metabolic activity of cell lines of the kidney of pig embryos carrying oncoviruses A and C (SPEV), rat glioma (C6), cancer of the human cervix (HeLa), and can be promising in medical practice for the treatment of cancer .

Compounds exhibiting cytotoxic activity against many types of tumor cells are known.

Such compounds include the alkaloid of a number of combretastatin A-4 and its derivatives, as well as 4-arylcoumarins and others. However, each of them has disadvantages.

The disadvantage of combretastatin A-4 (see, for example, US patent No. 4996237, CL IPC C07C 43/23, C07D 317/64, A61K 31/075, publ. 02.26.1991) and its derivatives (see application US No. 2009186857, according to IPC class A61K 31/66, published July 23, 2009) is low due to undesirable side effects when used in vivo due to their spontaneous conversion from the active cis form to an inactive trans form (Curr. Opin. Pharmacol. , 2001, 1,370).

4-Arylcoumarins exhibit low antiproliferacin and apoptosin-inducing activity compared with derivatives of combretastatin and colchicine (J. Med. Chem. 2003, 46, 5437; J. Med. Chem. 2011, 54, 3153).

A compound of diethyl (3,5-bis (arylidene) -4-oxopiperidin-1-yl) (aryl) methylphosphonate of the formula

where R ¹ = H, F, OMe; R ² = H, F,

possessing antiproliferative properties and low acute toxicity for the treatment of oncological diseases, including rhabdomyosarcoma, intestinal carcinoma, breast adenocarcinoma (see RF patent No. 2603194 according to IPC C07F 9/59, publ. 27.11.2016).

Chalcones exhibiting cytotoxic properties are also known (see Shih, H. et al. Rational design, synthesis and structure-activity relationships of antitumor (E) -2-benzylidene-l-tetralones and (E) -2-benzylidene-l-indanones // Bioorganic & medicinal chemistry letters. - 2000. - T. 10. - No. 5. - C. 487-490). Analysis of the antitumor activity of 2- (R-benzylidene) -3,4-dihydronaphthalen-1 (2H) -ones performed on a Jukart cell line (human T-lymphoblastic leukemia) showed that the concentration values of half-maximal inhibition of IC ₅₀ depend on the nature of the radical in aromatic ring.

So, the compounds of the formula

where R represents 3'-OCH ₃ , or 3'-Cl, or 3'-NO ₂ , or 3'-CH ₃ ; or 3 ', 5'-OCH ₃ , or 3', 5'-CH ₃ ; or 3 ', 5'-Cl, or 3', 5'-NO ₂ , showed good results. The value of IC ₅₀ ranged from 2 to 19 mm (depending on the radical).

In addition, the antitumor activity of the compounds 2- (R-benzylidene) -3,4-dihydronaphthalen-1 (2H) -ones, where R represents 3'-OCH ₃ , or 3'-Cl, or 3'-NO ₂ , or 3'-CH ₃ , 4'-OCH ₃ was also studied on other cell lines - on leukemic mouse cells P388 (lymphoid neoplasm of mice) and L 1210 (lymphatic leukemia), as well as on human cells Molt 4 / C8 and CEM T lymphocytes (T-lymphoblastic leukemia).

Substituted compounds are also known (see RF patent No. 2203883 according to IPC C07C 49/84, publ. 05/10/2003) of the general formula

where Ar is phenyl, which may be unsubstituted or substituted by one, two or three substituents independently selected from the group of Cl, Br, F, -Ое, NO ₂ , CF ₃ , С _1-4 lower alkyl, -NMe ₂ , - NEt ₂ , -SCH ₃ , -NHCOCH ₃ ; 2-thienyl, 2-furyl; 3-pyridyl; 4-pyridyl or 3-indolyl; R is —OCH ₂ R ₁ , where R _{1 is} selected from —CH = CME ₂ , —CMe = CH ₂ , —C≡CH; with the proviso that when Ar is phenyl, C ₄ -alkylphenyl, 4-methoxyphenyl or 3,4-dimethoxyphenyl, R may be any except 3-methyl-2-butenyloxy.

Mentioned compounds have antiproliferative activity against sensitive cancer cells as well as cells resistant to traditional chemotherapeutic drugs.

The invention is aimed at solving the problem of expanding the arsenal of drugs with cytotoxic effects that can be used as active components of antitumor drugs.

The claimed problem is solved in that the arylidene derivative of 3,4-dihydro-1 (2H) -naphthalenone having cytotoxic activity is a compound of the formula

The claimed compound: 4- (2,4-Dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid was synthesized by a one-stage technological method as a result of a three-way reaction 3.4 -dihydronaphthalen-1 (2H) -one, 2,4-dimethoxybenzaldehyde and 4-hydroxycoumarin (4-hydroxy-2H-chromen-2-one) in acetic acid in the presence of piperidine under microwave activation (power 600 W). The latter leads to the disclosure of the lactone fragment of the hybrid intermediate. This kind of transformation provides the basis for the reaction mechanism through the formation of 2- (2,4-dimethoxybenzylidene) -3,4-dihydronaphthalen-1 (2H) -one (the compound according to the first embodiment, where R ₁ = R ₂ = OCH3), which is basic for the formation of 4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid.

The invention is illustrated by drawings, which show graphs of the metabolic activity of cell lines when exposed to the claimed compounds, with:

- in FIG. 1 is a graph of metabolic activity of the C6 cell line;

- in FIG. 2 is a graph of the metabolic activity of the SPEV cell line;

- in FIG. 3 is a graph of the metabolic activity of the HELA cell line.

Obtaining a new compound and testing its cytotoxic activity is illustrated by the following example.

Example. Synthesis of 4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid.

To 1.19 g (7.5 mmol) of 2,4-dimethoxybenzaldehyde, 1.00 g (7.5 mmol) of 3,4-dihydronaphthalen-1 (2H) -one and 1.15 g (7.5 mmol) of 4-hydroxy-2H-chromen-2-one are added. To the resulting mixture was added 0.5 ml of glacial acetic acid for homogenization and 0.1 ml of piperidine catalyst. The reaction mixture is exposed to 600 W microwave radiation for 45 minutes. The resulting crystals are treated with hexane, washed with water and dried. 2.27 g (66.4%) of 4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid are obtained, mp. 50-51 ° C. ¹ H NMR spectrum (CDCl ₃ ), δ, ppm: 2.97 (t, 2H, CH _{2 (tetra)} ), 3.06 (t, 2H, CH _{2 (tetra)} ), 3.85 (s, 6H, OCH ₃ ), 5.36 (s. 1H, OH), 5.61 (s. 1H, CH), 6.45-8.22 (m. 11H, Ar). Found,%: C 73.39, H 5.41. C ₂₈ H ₂₄ O ₆ . Calculated,%: C 73.67, H 5.30.

The cytotoxicity of 4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid was determined using the MTT test on kidney cell cultures of pig embryos, carrying oncoviruses A and C (SPEV), rat glioma cells (C6), and human cervical cancer (HeLa). The MTT test is based on the ability of living cells to reduce yellow 3- (4,5-dimethylthiazol-2-yl) -2,5-tetrazolium bromide (MTT) into the purple-blue intracellular MTT-formazan crystals soluble in DMSO. The decrease in the optical density of the experimental samples compared with the control, recorded on a flat-bed spectrophotometer, indicates the cytotoxic effect of the studied substances on the cells.

Cells of the SPEV, C6 (glioma) and HeLa clone 11 lines suspended in culture medium were added to the wells of 96-well polystyrene plates (200 μl of medium and approximately 50,000 cells of the corresponding line were added to one well). The culture medium was DMEM supplemented with 10% fetal calf serum, as well as antibiotics penicillin (50 units / ml) and streptomycin (50 μg / ml) to prevent bacterial contamination. The cultures were grown in an atmosphere of a CO ₂ incubator for 2 days until a monolayer of cells with a 70% saturation density was formed.

Test substances were dissolved in DMSO to the highest concentration. The maximum concentration of compound 1 was 55.56 mg / ml, compounds 2-50 mg / ml, and compounds 3-52.63 mg / ml.

After 48 hours of incubation, various concentrations of the test compounds were added to the cell cultures by the method of rasterization. Each concentration was applied in triplicate. A 1: 100 dilution in culture medium was used as the initial dilution of the samples. DMSO solvent at a concentration of 1% was added to control wells. Compound plates were again placed in a CO ₂ incubator for 24 hours.

To determine respiratory activity, MTT reagent was dissolved in buffered saline to a concentration of 1 mg / ml.

After 72 hours of incubation of the cell lines with the tested substances, the wells of the plates were freed from the medium and 100 μl of MTT solution was added to them. The plates were placed for 1 hour in a CO ₂ incubator, then the liquid from the wells was removed, 200 μl of DMSO solution was added to them and incubated for 10 min at 37 ° C in an incubator.

Using a tablet format spectrophotometer, the optical density of the obtained samples was determined at a wavelength of 540 nm, and the measured background absorption at a wavelength of 690 nm was subtracted. The respiratory activity of the control samples was taken as 100%.

In the Table and in FIG. 1, 2, 3 presents the results of determining the metabolic activity of cell lines C6, SPEV and HeLa when exposed to the claimed compounds in the maximum concentration used.

From the above data it follows that the claimed compound 4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid in the maximum concentration used suppresses metabolic activity of cell lines C6, SPEV and HeLa

Thus, the claimed compound can be promising for the creation of drugs aimed at the treatment of cancer.

Claims (2)

4- (2,4-dimethoxyphenyl) -2- (2-hydroxyphenyl) -5,6-dihydro-4H-benzo [h] chromene-3-carboxylic acid having cytotoxic activity, of the formula

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