JPH06122623A - Antitumor agent - Google Patents

Abstract

(57) [Summary] (Modified) [Objective] To provide an antitumor agent containing a chalcone compound as an active ingredient and having a tumor cell growth inhibitory action and a cancer promoter inhibitory action. [Structure] An antitumor agent containing a chalcone compound of the following formula as an active ingredient. [In the formula, R represents —OH, —CH ₃ ; R ₁ represents H, —OH, —
OCH _3, a -CH _3; R ₂ represents respectively H, a -CH _3. ]

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an antitumor agent, and more specifically to an antitumor agent containing a chalcone compound as an active ingredient and having a tumor cell growth inhibitory action and a carcinogenic promoter inhibitory action. .

[0002]

BACKGROUND OF THE INVENTION Flavonoids present in crude drugs (for example, quercetin, apigenin, liquiritigenin, etc.)
However, it has been reported to have a tumor cell growth inhibitory action or a cancer promoter inhibitory action.

However, these compounds are extracted from crude drugs,
It must be separated and further purified. In addition, when trying to obtain it by chemical synthesis, several steps are required,
Depending on the synthetic method, it is necessary to make a chalcone derivative once and then close it.

[0004]

As described above, in order to obtain a flavonoid compound by the conventional method, complicated work or multi-step process is required.

The present invention has been made from the above viewpoint, and an object thereof is to obtain an effective compound in a short process without requiring complicated work.

[0006]

[Means for Solving the Problems] In order to solve the above problems, the present inventor has searched for a substance having an antitumor activity which is present in a herbal medicine, and in the process of lycochalcone contained in Shinji licorice extract. As a result of finding that A is effective and further searching for chalcone which is more effective and easier to synthesize with its related compound, we succeeded in developing an effective compound and reached the present invention.

That is, the present invention is an antitumor agent containing the chalcone compound represented by Chemical formula 1 as an active ingredient, and an antitumor agent containing lycochalcone A represented by Chemical formula 8 as an active ingredient.

The present invention will be described in detail below. <1> Chalcone Compound Used in the Present Invention As described above, the antitumor agent of the present invention contains the chalcone compound represented by Chemical formula 9 or lycochalcone A as an active ingredient.

[0009]

[Chemical 9]

However, in the formula, R represents --OH or --CH ₃ and R ₁
Is -H, -OH, -OCH _3, one of the -CH _3, R
₂ respectively represent -H or -CH _3. Examples of the chalcone compound include 3-hydroxychalcone represented by Chemical formula 2, 3,3′-dihydroxychalcone represented by Chemical formula 3, 3′-methoxy-3-hydroxychalcone represented by Chemical formula 4, and Chemical formula 5 3'-methyl-3-hydroxychalcone represented by the formula, 3'-hydroxy-3 represented by the formula 6
-Methylchalcone, 3-hydroxy represented by Chemical formula 7
4'-methyl chalcone and the like can be mentioned.

The chalcone compound and lycochalcone A used in the present invention can be provided as salts thereof, if necessary. For example, sodium salt, potassium salt and the like can be exemplified.

3'-Methyl-3-hydroxychalcone and 3'-hydroxy-3-methylchalcone are novel compounds. The physical properties of 3'-methyl-3-hydroxychalcone are shown below.

Melting point: 106-107 ° C. Mass spectrum% Int m / z 238 (M ⁺ ) (base) m / z 237 (59.0) m / z 147 (22.0) m / z 119 (28.3) High-resolution mass spectrum calculation Value: 238.0994 Measured value: 238.0994

<2> Method for Producing Chalcone Compound Used in the Present Invention All the above chalcone compounds can be obtained by chemical synthesis by a conventional method. The manufacturing method is shown below.

As shown in Chemical formula 10, the chalcone compound can be obtained by Claisen-Schmidt condensation of acetophenone having a functional group necessary for the target compound and benzaldehyde having a functional group necessary for the target compound.
Depending on the type of the functional group, the functional group may be protected and condensed if necessary, and then deprotected to obtain the target compound.

[0016]

[Chemical 10]

However, R ¹ represents —H, —OH, —OCH ₃ or —CH ₃ , and R ² represents —H, —OH or —CH ₃ . Next, a production example of 3′-methyl-3-hydroxychalcone will be shown as an example of the production method.

5 g of m-hydroxybenzaldehyde was dissolved in 150 ml of acetonitrile, and further 1 g of 18-crown-6-ether and 5 m of chlorodimethyl ether.
l was added, and the mixture was stirred at room temperature for a while, 25 g of anhydrous potassium carbonate was added with stirring, and the reaction was continued for 1 hour.

After filtering the reaction solution, the filtrate is concentrated under reduced pressure,
The mixture was partitioned with diethyl ether-water, and the ether layer was separated. The obtained ether layer was dried with sodium sulfate for 6 hours, sodium sulfate was removed by filtration, and the solvent was distilled off at 40 ° C or lower. The obtained residue was chromatographed on silica gel (solvent hexane: ethyl acetate = 9:
Separation according to 1) gave 5.5 g of m-methoxymethoxybenzaldehyde.

5.5 g of m-methoxymethoxybenzaldehyde thus obtained and 4.5 g of m-methylacetophenone were dissolved in 40 ml of ethanol, and further dissolved in 25 ml.
% Potassium hydroxide aqueous solution 20ml, at room temperature 24
Stir for hours.

After the reaction was completed, the reaction solution was neutralized with IN HCl, concentrated under reduced pressure, partitioned with diethyl ether-water, and the ether layer was dried with sodium sulfate for 6 hours. After drying, the sodium sulfate was removed by filtration, and the solvent was added to 4
After distilling off at 0 ° C. or lower, the residue was subjected to silica gel column chromatography (developing solvent hexane: ethyl acetate = 4:
Separation by 1) gave 3-methoxymethoxy-3'-methylchalcone.

3 g of the obtained 3-methoxymethoxy-3'-methylchalcone was dissolved in 60 ml of methanol to obtain 3N.
-HCI 20ml was added and it boiled and refluxed for 30 minutes. After completion of the reaction, the reaction solution was concentrated under reduced pressure, partitioned with diethyl ether-water, and the ether layer was dried over sodium sulfate.

After sodium sulfate was removed by filtration and the solvent was distilled off, the residue was dissolved in a small amount of methanol and subjected to high performance liquid chromatography (solvent methanol: water = 75: 25) using an octadecylsilyl (ODS) column. I collected it. Detection of the chalcone compound was performed by measuring the absorbance at 254 nm. By recrystallizing the obtained fraction with methanol: water, 3'-methyl-
1.5 g of 3-hydroxychalcone was obtained.

<3> Antitumor Agent of the Present Invention The antitumor agent of the present invention contains, as an active ingredient, one or more selected from the above chalcone compound or lycochalcone A, and its formulation and manufacturing method are particularly Regardless, it is manufactured by a conventional method.

For example, one or several non-toxic pharmaceutically acceptable excipients such as lactose, potato starch,
It can be made into powders, granules, tablets, capsules and the like in which calcium carbonate, sodium alginate or the like is dispensed. Also, when preparing an injection, the solvent is distilled water for injection,
Alternatively, polyethylene glycol or the like is used, or a dispersant may be added thereto.

The method of administration may be oral or parenteral. The dose varies depending on the age, symptoms, etc. of the patient, but generally, by oral administration, the desired effect can be expected by using the amount of the chalcone compound in the range of 0.01 to 100 mg per adult per day. . In the case of parenteral administration, 60% of the above oral administration per adult per day is appropriate.

[0027]

The action of the chalcone compound used in the present invention will be described below. <Tumor Cell Proliferation Inhibitory Action> The tumor cell proliferation inhibitory action was evaluated by using as an index whether the chalcone compound and lycochalcone A used in the present invention inhibit the proliferation of various tumor cells in vitro.

(1) Effect on gastric cancer Human gastric cancer HGC-27 cells were sown on a dish having a diameter of 3.5 cm and cultured, and after 1 day of culturing, the cells were transformed into this culture medium.
Each of the chalcone compounds represented by ~ 7 or lycochalcone A was added so as to have a concentration of 1 µg / ml.

After culturing for 3 days, the number of cells was measured,
The growth inhibition rate was calculated. The results are shown in Table 1.

[0030]

[Table 1]

It was revealed that each of the chalcone compounds exhibited a strong antiproliferative effect. Among them, 3'-methyl-3-hydroxychalcone was most effective.

(2) Effect of 3'-methyl-3-hydroxychalcone on proliferation of HGC-27 cells HGC-27 cells were seeded on a dish having a diameter of 3.5 cm,
After 1 day of culture, various concentrations of 3'-methyl-3-hydroxychalcone were added, and the cells were further cultured for 1 to 4 days, and then the number of cells was measured. The results are shown in Fig. 1.

It was found that 3'-methyl-3-hydroxychalcone suppressed the proliferation of HGC-27 cells in a concentration-dependent manner, and the ID _{50 on the} third day after the treatment was about 0.5 μg / ml ( (Fig. 1).

The chalcone compound completely suppressed the proliferation of HGC-27 cells even at a concentration of 1 μg / ml, and its efficacy was continued for at least 4 days (FIG. 2).

(3) 3'- for various human tumor cells
Action of methyl-3-hydroxychalcone HeLa cells derived from cervical cancer, GOTO derived from neuroblastoma
Cells and pancreatic cancer-derived PANC-1 cells with a diameter of 3.5 cm
4 × 10 ⁴ cells were seeded on each dish and cultured.
After 1 day, the compound (4) was added to each culture solution to a concentration of 1 μg /, and the culture was continued for 3 days to measure the number of cells. The results are shown in Table 2.

[0036]

[Table 2]

From the above results, it was found that 3'-methyl-3-hydroxychalcone exhibited a growth inhibitory effect on any human tumor cells.

<2> Inhibitory effect of carcinogenic promoter Various substances are known as carcinogenic promoters, and among them, 12-O-tetradecanoylphorbol-1.
3-acetate (TPA) is one of the most powerful.

Although the entire mechanism of action of TPA is still unknown at present, the enhancement of phospholipid metabolism in cells is considered to be one of the important factors leading to carcinogenic promotion, and the action of TPA It has been clarified that many of the compounds that suppress the tumorigenesis block the carcinogenic promotion by TPA and efficiently reduce the tumorigenesis.

Therefore, as an index of the anti-promoter action of the chalcone compound developed this time, the efficacy of suppressing the phospholipid metabolism promotion of cells by TPA was measured. The effect of chalcone compounds on the inhibition of phospholipid synthesis enhancement of cells caused by TPA
The test was performed in vitro.

The test compound was added to the HeLa cell culture medium at a concentration of 5 μg / ml. For the control, only the same amount of solvent used in the main experiment was added. After 1 hour, TPA (50 nM) and radioactive inorganic phosphate ³² Pi were added, and the culture was continued for another 4 hours. Then, the phospholipids of the cells were extracted, the radioactivity was measured, and it was calculated how much the radioactivity uptake promoted by TPA was suppressed.
The results are shown in Table 3.

[0042]

[Table 3]

It was revealed that all chalcone compounds suppressed the ³² Pi uptake promoted by TPA. Among them, 3'-methyl-3-hydroxychalcone was found to have a particularly strong antipromoter activity.

[0044]

EXAMPLES Examples of the present invention will be described below.

[0045]

[Formulation Example 1] Tablet A tablet containing 5 mg of chalcone compound per tablet is produced as follows. (Production method) Crystals of the chalcone compound are crushed, and lactose and starch are added thereto and mixed. 10% starch paste is added to the above mixture and stirred to produce granules. Particle size after drying is about 85
Granulate to 0 micron, mix with talc and magnesium stearate and tablet.

[0046]

[Formulation Example 2] Capsule A capsule containing 20 mg of chalcone compound per capsule is produced as follows. (Manufacturing method) The chalcone compound is well pulverized, starch, lactose and magnesium stearate are added, mixed well, and then filled in a capel.

[0047]

[Formulation Example 3] Injection solution An injection solution is produced as follows. (Manufacturing method) The chalcone compound is dissolved in distilled water containing Macrogol 4000 and a prescribed amount of sodium chloride to adjust the pH.
After adjusting to around 7.0, fill and seal the ampoule.

[0048]

The antitumor agent of the present invention has both a strong tumor cell growth inhibitory effect and an anticarcinogenic promoter activity. This effect enables comprehensive measures against cancer such as prevention and treatment of cancer.

[Brief description of drawings]

FIG. 1 is a graph showing changes in tumor cell inhibitory effect on chalcone compound concentration.

FIG. 2 is a diagram showing suppression of tumor cell growth in the presence of a chalcone compound.

Claims (8)

[Claims] 1. An antitumor agent containing the chalcone compound represented by Chemical formula 1 as an active ingredient. [Chemical 1] However, in the formula, R is —OH or —CH ₃ , and R ₁ is —H, —O.
H, -OCH _3, one of -CH _3, R ₂ represents respectively -H or -CH _3. 2. The antitumor agent according to claim 1, wherein the chalcone compound is 3-hydroxychalcone represented by Chemical formula 2. [Chemical 2] 3. The antitumor agent according to claim 1, wherein the chalcone compound is 3,3′-dihydroxychalcone represented by Chemical formula 3. [Chemical 3] 4. The antitumor agent according to claim 1, wherein the chalcone compound is 3′-methoxy-3-hydroxychalcone represented by Chemical formula 4. [Chemical 4] 5. The antitumor agent according to claim 1, wherein the chalcone compound is 3′-methyl-3-hydroxychalcone represented by Chemical formula 5. [Chemical 5] 6. The antitumor agent according to claim 1, wherein the chalcone compound is 3′-hydroxy-3-methylchalcone represented by Chemical formula 6. [Chemical 6] 7. A chalcone compound represented by Chemical formula 3
The antitumor agent according to claim 1, which is hydroxy-4'-methylchalcone. [Chemical 7] 8. An antitumor agent containing Lycochalcone A represented by Chemical formula 8 as an active ingredient. [Chemical 8]