JPH06279432A - New compound and anticancer agent containing the compound as active component - Google Patents

Abstract

(57) [Summary] [Object] An object of the present invention is to provide an anticancer agent having few side effects and a strong anticancer effect. The present invention includes the following formula I (In the formula I, R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ₄ represents an oxygen atom, a sulfur atom or NH.) And a pharmaceutically acceptable salt thereof, and an anticancer agent comprising the compound of formula I and a pharmaceutically acceptable salt thereof as an active ingredient.

Description

Detailed Description of the Invention

FIELD OF THE INVENTION The present invention relates to a novel compound and an anticancer agent containing the compound as an active ingredient.

[0002]

2. Description of the Related Art Currently, clinically used drugs are alkylating agents typified by cyclophosphamide, folate antagonists typified by methotrexate, and purine antagonists typified by 6-mercaptopurine. , Pyrimidine antagonists typified by fluorouracil, plant alkaloids, bacterial components, platinum complexes and the like.

However, these drugs have advantages and disadvantages in terms of side effects, administration forms and administration methods, and at present, satisfactory drugs have not been developed for the treatment of cancer.

Therefore, there are few side effects and a strong anti-cancer effect,
The development of an ideal drug has been desired.

[0005]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors succeeded in obtaining a compound represented by the following formula I and a pharmaceutically acceptable salt thereof. , As a result of examining these pharmacological actions,
It was confirmed that it showed a remarkable anti-cancer effect, had low toxicity and high safety.

That is, the present invention provides the following formula I (In the formula I, R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ₄ represents an oxygen atom, a sulfur atom or NH.) And a pharmaceutically acceptable salt thereof (hereinafter referred to as the compound of the present invention) and an anticancer agent containing the compound of the present invention as an active ingredient.

The compound of the present invention is a novel compound and has an excellent anti-cancer effect, especially an excellent effect on liver cancer. It is a compound that can solve the conventional problems because it can be easily synthesized and has low toxicity.

The compound of the present invention can be easily obtained by the following method using gallic acid (garlic acid) as a lead substance.

That is, the hydroxyl group of garlic acid is protected by, for example, methoxymethylation, and then hydrolyzed to a carboxylic acid. Next, by condensation with disesamol, 3,4-methylenedioxyaniline, 3,4-methylenedioxythiophenol, etc., an ester form, an amide form, and a thioester form are obtained. The compound of the invention is obtained.

If necessary, the compound of the present invention can be obtained by alkylation according to a conventional method using methyl iodide, ethyl iodide and the like.

Hydrolysis may be carried out by a conventional method, for example, sodium hydroxide is reacted in methanol.

In the condensation reaction, diethyl chlorophosphate, dicyclohexylcarbodiimide (DCC) or the like is used as a condensing agent, and it is preferably carried out in the presence of a base. As the base, triethylamine or dimethylaminopyridine (DMAP) is appropriately selected.

In the deprotection reaction, for example, in the case of demethoxymetholation, it can be easily carried out by making acidic conditions with hydrochloric acid.

The garlic acid may be, for example, one commercially available from Wako Pure Chemical Industries, Ltd., or may be isolated from a garlic acid-containing crude drug such as Hishimi [Reference: Pharmaceutical Journal, 106 ( 2), 183-185 (1986)].

Next, the fact that the compound of the present invention has an excellent anticancer activity will be described with reference to experimental examples.

Experimental Example 1 The compound of the present invention was dissolved in dimethyl sulfoxide (DMSO) so as to have a concentration of 20 mg / ml and stored at -4 ° C.

The dRLh-84 cells were 96 at a concentration of 3 × 10 ⁴ cells / ml.
-0.1 ml / wel in a well microplate (Falcon)
l sprinkled each.

After pre-culture for 24 hours, the plate was washed with PBS and 20 mg /
Add 25 μl of ml of the compound of the invention to 4975 μl of medium (100 μl
(g / ml, 0.5% DMSO), final concentration 0 ~ by serial dilution
A medium of 100 μg / ml was prepared and 0.1 ml / well was added to each medium.

After culturing for 48 hours, the medium was washed with 0.1 m of medium.
l / well and MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphe
Nyltetrazolium bromide, manufactured by SIGMA) was dissolved in 5 mg / ml phosphate-buffered saline and sterilized with a filter, and 10 μl / well was added to each.

After culturing for 6 hours, 10% sodium dodecyl sulfate
(sodium dodecyl sulfate, SDS) was added by 0.1 ml / well, and after culturing for 18 hours, the absorbance was measured at 577 nm and 655 nm (MICROPLATE READER Model 450, manufactured by Bio Rad). All the cultures were performed at 37 ° C and 5% carbon dioxide.

The ratio of color development of the garlic acid or the compound of the present invention to the group of the compound of the present invention to which 0 μg / ml garlic acid or the compound of the present invention was calculated by subtracting the absorbance of the medium alone was calculated, and 50% of the garlic acid or the compound of the present invention was calculated. The effective concentration (ED ₅₀ , μg / ml) was determined.

The results are shown in Table 1.

Table 1

From the above results, it was confirmed that the compound of the present invention has an excellent anticancer activity. In particular, the compounds obtained in Examples 1 and 3 were 2 to 3 times more active than the garlic acid as the lead substance.

That is, although the compound of the present invention is derived from garlic acid as a raw material, it has extremely high activity as compared with garlic acid, and there is a problem in terms of absorption with garlic. It has an advantage that absorption is improved as compared with acid.

Next, the dose and formulation of the compound of the present invention will be explained.

The compound of the present invention can be administered to animals and humans as it is or together with a conventional pharmaceutical carrier. The dosage form is not particularly limited and may be appropriately selected and used as needed, and examples thereof include oral preparations such as tablets, capsules, granules, fine granules and powders, parenteral preparations such as injections and suppositories. To be

In order to exert a desired effect as an oral preparation, it varies depending on the age, body weight and degree of disease of a patient, but usually 10 mg to 500 mg as the weight of the compound of the present invention is used in adults.
It seems appropriate to take it several times a day.

Oral preparations include, for example, starch, lactose, sucrose,
Mannitol, carboxymethyl cellulose, corn starch, inorganic salts and the like are used in a conventional manner.

In addition to the above-mentioned excipients, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a flavoring agent, a coloring agent, a flavoring agent, etc., may be appropriately used in this type of preparation. You can
Specific examples of each are as follows.

[Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch, methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose,
Ethyl cellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low-substituted hydroxypropyl cellulose.

[Surfactant] Sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 8
0.

[Lubricant] Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol.

[Flowability Accelerator] Light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The compounds of the present invention can also be administered as suspensions, emulsions, syrups and elixirs, and these various dosage forms may contain flavoring agents and coloring agents. Good.

In order to exert the desired effect as a parenteral agent, it depends on the age, body weight and degree of disease of the patient.
The weight of the compound of the present invention is usually 0.1 mg to 10 per day in adults.
Intravenous injections up to mg, intravenous drip, subcutaneous injection, and intramuscular injection seem appropriate.

This parenteral preparation is manufactured by a conventional method, and is generally used as a diluent in distilled water for injection, physiological saline, glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol. Etc. can be used. Further, if necessary, a bactericide, a preservative, and a stabilizer may be added. Further, from the viewpoint of stability, this parenteral preparation may be filled in a vial or the like and then frozen, the water content may be removed by a usual freeze-drying technique, and a liquid preparation may be re-prepared from the freeze-dried product immediately before use. Further, an isotonicity agent, a stabilizer, a preservative, a soothing agent and the like may be added as needed.

Other parenteral agents include external preparations, coating agents such as ointments, and suppositories for rectal administration.
It is manufactured according to a conventional method.

Next, specific examples of the production of the compound of the present invention will be shown as examples, but the present invention is not limited thereto.

Example 1 Gallic acid (garlic acid) 50 was placed in a 2 l eggplant-shaped flask.
g, 500 ml of anhydrous dimethylformamide (DMF), 412 ml of diisopropylethylamine, and after purging with argon,
158 ml of methoxymethyl chloride was added dropwise under ice cooling, and the mixture was stirred at room temperature overnight. This reaction solution was poured into 2 l of ice water, weakly acidified with diluted hydrochloric acid (pH 3-4), and then extracted twice with ethyl acetate (1 l x 2),
The ethyl acetate layer was washed once with 5% sodium hydroxide and water, dried (MgSO ₄ ), and the solvent was evaporated under reduced pressure to give a yellow oil.

To this was added 500 ml of methanol and 500 ml of 2N sodium hydroxide, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated under reduced pressure (removal of methanol), washed once with ethyl acetate, the aqueous layer was made weakly acidic (pH 4) with 2N hydrochloric acid under ice cooling, and the white insoluble matter precipitated was collected by filtration (with water). Wash well), dry (60 ° C / 5
mmHg) to obtain 61.7 g of colorless crystals. This was recrystallized from ethyl acetate to obtain 60.8 g of 3,4,5-trimethoxymethylbenzoic acid having the following physicochemical properties as colorless needle crystals.

Melting point: 149-151 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 2964,29
12,2828,2640,1690,1596,1502,1434,1394,1328,1304,11
94,1156,1112,1054,954,920,868,762,712,580,546 Proton Nuclear Magnetic Resonance Spectra (δ ppm in CDCl ₃ ):
3: 3.53 (6H, s), 3.63 (3H, s), 5.24 (4H, s), 5.26 (2H, s), 7.60
(2H, s) Mass spectrum (EI-MS) m / z (%): 302 (M ⁺ , 2), 226 (16), 209
(6), 196 (7), 45 (100)

In a 100 ml round-bottomed flask equipped with a calcium chloride tube, 4.0 g of 3,4,5-trimethoxymethoxybenzoic acid obtained in, 3.8 g of dimethylaminopyridine, and methylene chloride.
To the solution containing 40 ml, under ice cooling, 2.0 ml of diethyl chlorophosphate was added, and the mixture was stirred at room temperature for 30 minutes, and then sesamol 1.7 was added.
4 g was added, and the mixture was stirred at room temperature for 4 hours. Water was added to the reaction solution, extracted twice with ethyl acetate (200 ml × 2), and the ethyl acetate layer was 5%.
Hydrochloric acid, 5% sodium hydroxide, water, saturated saline solution
It was washed once, dried (MgSO ₄ ), and the solvent was evaporated under reduced pressure to give a brown solid. This was recrystallized from ethyl acetate-n-hexane to give 3,4-methylenedioxyphenyl having the following physicochemical properties.
4.9 g of 3,4,5-trimethoxymethoxybenzoate was obtained as colorless needle crystals.

Melting point: 79-80 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 2956,29
36,2908,2828,1728,1598,1504,1486,1444,1436,1392,13
26,1302,1218,1176,1160,1110,1044,962,950,918,878,7
92,760,700,600 Proton Nuclear Magnetic Resonance Spectrum (δ ppm in CDCl ₃ ):
3.53 (6H, s), 3.63 (3H, s), 5.25 (2H, s), 5.27 (4H, s), 6.00 (2
H, s), 6.62 (1H, dd, J = 8.3,2.0Hz), 6.70 (1H, d, J = 2.0Hz), 6.
81 (1H, d, J = 8.3Hz), 7.66 (2H, s) Mass spectrum (EI-MS) m / z (%): 422 (M ⁺ , 2), 285 (35), 209
(84), 179 (14), 45 (100)

To a 100 ml eggplant-shaped flask, 3.0 g of 3,4-methylenedioxyphenyl 3,4,5-trimethoxymethoxybenzoate and 30 ml of 35% hydrochloric acid-methanol (1: 6 Vol / Vol) were placed, and the mixture was kept at room temperature for 2 hours. It was stirred. Pour this reaction solution into ice water,
It was extracted twice with ethyl acetate (150 ml × 2), the ethyl acetate layer was washed twice with saturated brine and dried (MgSO ₄ ), and the solvent was evaporated under reduced pressure to give a white solid. This was recrystallized from ethyl acetate-n-hexane to obtain 1.95 g of colorless fine needle crystals of 3,4-methylenedioxyphenyl 3,4,5-trihydroxybenzoate having the following physicochemical properties.

Melting point: 228-230 Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 3500, 34
20,1710,1610,1536,1492,1386,1312,1220,1168,1040,96
2,930,854,794,754 Proton Nuclear Magnetic Resonance Spectra (δ ppm in CD ₃ OD):
5.99 (2H, s), 6.60 (1H, dd, J = 8.3,2.2Hz), 6.70 (1H, d, J = 2.2
Hz), 6.83 (1H, d, J = 8.3Hz), 7.17 (2H, s)

Example 2 In a 200 ml eggplant-shaped flask equipped with a calcium chloride tube,
3,4,5-trimethoxymethoxybenzoic acid 4.0 g obtained in Example 1, a solution of triethylamine 4.4 ml, methylene chloride 40 ml under ice cooling, diethyl chlorophosphate 2.0 m
l was added and stirred at room temperature for 1 hour, 1.73 g of 3,4-methylenedioxyaniline was added, and the mixture was stirred at room temperature overnight. Water was added to this reaction solution, extracted twice with ethyl acetate (200 ml × 2), the ethyl acetate layer was washed once with 5% hydrochloric acid, 5% sodium hydroxide, water, and saturated saline, respectively, and dried (MgSO ₄ ), The solvent was distilled off under reduced pressure to obtain a white solid. This was recrystallized from ethyl acetate-n-hexane to give 4.4 g of N- (3,4-methylenedioxyphenyl) -3,4,5-trimethoxymethoxybenzamide having the following physicochemical properties as colorless needles. Got as.

Melting point: 115-117 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 3260,29
04,1644,1590,1530,1494,1436,1324,1220,1160,1112,10
58,922,856,764 Proton nuclear magnetic resonance spectrum (δ ppm in CDCl ₃ ):
3.52 (6H, s), 3.62 (3H, s), 5.21 (2H, s), 5.26 (4H, s), 5.97 (2
H, s), 6.77 (1H, d, J = 8.3Hz), 6.89 (1H, dd, J = 8.3,2.0Hz), 7.
33 (1H, d, J = 2.0Hz), 7.33 (2H, s) Mass spectrum (EI-MS) m / z (%): 421 (M ⁺ , 30), 374 (23), 345
(23), 209 (100), 165 (15), 137 (12), 45 (96)

In a 100 ml round-bottomed flask, 3.0 g of N- (3,4-methylenedioxyphenyl) -3,4,5-trimethoxymethoxybenzamide obtained in Example 2, 35% hydrochloric acid-methanol
(1: 6 Vol / Vol) (30 ml) was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was poured into ice water and extracted twice with ethyl acetate (150 ml ×
2), the ethyl acetate layer was washed twice with saturated saline and dried (MgS
O _4), the solvent to obtain a distilled off under reduced pressure to a white solid. This was recrystallized from ethyl acetate-n-hexane to give 1.78 g of N- (3,4-methylenedioxyphenyl) -3,4,5-trihydroxybenzamide having the following physicochemical properties as colorless fine needle crystals. Got as.

Melting point: 267-270 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 3416, 33
12,1668,1614,1534,1492,1450,1336,1198,1034,930,86
0,792,744 Proton nuclear magnetic resonance spectrum (δ ppm in CDCl ₃ ):
5.93 (2H, s), 6.77 (1H, d, J = 8.8Hz), 6,93 (2H, s), 6.99 (1H, d
d, J = 8.8,2.2Hz), 7.24 (1H, d, J = 2.2Hz) Mass spectrum (EI-MS) m / z (%): 289 (M ⁺ , 24), 153 (24), 137
(100), 79 (28)

Example 3 In a 200 ml eggplant-shaped flask equipped with a calcium chloride tube,
3,4,5-trimethoxymethoxybenzoic acid 4.0 g obtained in Example 1, a solution of triethylamine 4.4 ml, methylene chloride 40 ml under ice cooling, diethyl chlorophosphate 2.0 m
l was added, and the mixture was stirred at room temperature for 1 hr, 3,4-methylenedioxythiophenol (1.94 g) was added, and the mixture was stirred at room temperature overnight. Water was added to this reaction solution, and extracted twice with ethyl acetate (200 ml × 2),
The ethyl acetate layer was washed once with 5% hydrochloric acid, 5% sodium hydroxide, water and saturated brine, dried (MgSO ₄ ), and the solvent was evaporated under reduced pressure to give a white solid. This was recrystallized from ethyl acetate-n-hexane to give 4.63 g of S- (3,4-methylenedioxyphenyl) 3,4,5-trimethoxymethoxybenzoic acid thioester having the following physicochemical properties as colorless needles. Obtained as crystals.

Melting point: 62-64 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 2904,16
80,1592,1500,1480,1434,1318,1240,1160,1110,1054,91
8,810,772,686 Proton nuclear magnetic resonance spectrum (δ ppm in CDCl ₃ ):
3.52 (6H, s), 3.62 (3H, s), 5.23 (2H, s), 5.26 (4H, s), 6.03 (2
H, s), 6.87 (1H, d, J = 8.0Hz), 6.94 (1H, d, J = 1.7Hz), 6.98 (1
H, dd, J = 8.0,1.7Hz), 7.52 (2H, s) Mass spectrum (EI-MS) m / z (%): 438 (M ⁺ , 5), 362 (3), 315
(4), 285 (100), 241 (25), 209 (49), 165 (32)

In a 100 ml round-bottomed flask, 3.0 g of S- (3,4-methylenedioxyphenyl) 3,4,5-trimethoxymethoxybenzoic acid thioester obtained in Example 3 was added, and 35% hydrochloric acid-methanol (1: (6 Vol / Vol) 30 ml was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was poured into ice water and extracted twice with ethyl acetate (150 m
l × 2), the ethyl acetate layer was washed twice with saturated saline and dried (MgS
O _4), the solvent to obtain a distilled off under reduced pressure to a white solid. This was recrystallized from ethyl acetate-n-hexane to obtain 1.97 g of 3,4-methylenedioxyphenyl 3,4,5-trihydroxybenzoic acid thioester having the following physicochemical properties as colorless fine phosphorus-like crystals. .

Melting point: 217-220 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , KBr): 3340,16
44,1596,1476,1326,1238,1142,1036,1012,838,760,690,
604 Proton nuclear magnetic resonance spectrum (δ ppm in CD ₃ OD):
6.02 (2H, s), 6.88 (1H, d, J = 7.8Hz), 6.90 (1H, d, J = 1.7Hz),
6.95 (1H, dd, J = 7.8,1.7Hz), 7.06 (2H, s) Mass spectrum (EI-MS) m / z (%): 306 (M ⁺ , 5), 153 (100), 125
(14), 96 (21), 58 (17)

Next, the formulation examples of the present invention will be described.

[0057]

According to the above formulation, the ingredients (1) to (2) were uniformly mixed and compression-molded with a tableting machine to give tablets (200 mg each).

Each tablet contains 20 mg of the compound obtained in Example 1, and 3 to 10 tablets for adults are to be taken in several divided doses per day.

[Formulation Example 2] Crystalline cellulose 84.5 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Compound obtained in Example 2 10 g Total 100 g

According to the above formulation, a part of and was uniformly mixed, compression-molded, crushed, and the remaining amount of and was mixed, and the mixture was compression-molded with a tableting machine to give one tablet.
0 mg tablets were obtained.

Each tablet contains 20 mg of the compound obtained in Example 2, and 3 to 10 tablets for adults are to be taken in several divided doses per day.

[Formulation Example 3] Crystalline cellulose 79.5 g 10% Hydroxypropyl cellulose ethanol solution 50 g Carboxymethyl cellulose calcium 5 g Magnesium stearate 0.5 g Compound 10 g obtained in Example 3 Total 145 g

According to the above formulation, and were uniformly mixed, the mixture was kneaded by a conventional method, granulated by an extrusion granulator, dried and crushed, and then mixed, and compressed by a tableting machine. It was molded to obtain a tablet of 200 mg each.

Each tablet contains 20 mg of the compound obtained in Example 3, and 3 to 10 tablets for adults are to be taken in several divided doses per day.

[0066]

According to the above formulation, the components (1) to (4) are uniformly mixed, compression-molded by a compression molding machine, and then crushed by a crusher,
Sieve to obtain granules.

1 g of this granule contains 100 mg of the compound obtained in Example 1, and 1 to 2 g of an adult is taken in several divided doses per day.

[Formulation Example 5] Crystalline cellulose 86.5 g 10% Hydroxypropyl cellulose ethanol solution 35 g Compound obtained in Example 2 10 g Total 131.5 g

According to the above-mentioned recipe, the ingredients (1) to (4) were uniformly mixed and kneaded. After granulating with an extrusion granulator, it was dried and sieved to obtain granules.

1 g of this granule contains 100 mg of the compound obtained in Example 2, and 1 to 2 g of an adult is taken in several divided doses per day.

[Formulation Example 6] Cornstarch 89.5 g Light anhydrous silicic acid 0.5 g Compound obtained in Example 3 10 g Total 100 g

According to the above-mentioned formulation, ~ was uniformly mixed, and 200 mg was filled in a No. 2 capsule.

One capsule of this capsule contains 20 mg of the compound obtained in Example 3, and 3 to 10 capsules for an adult are to be taken in several divided doses per day.

[Formulation Example 7] Distilled water for injection 89.5 g Soybean oil 5 g Soybean phospholipid 2.5 g Glycerin 2 g Compound obtained in Example 1 1 g Total amount 100 g

According to the above-mentioned prescription, was dissolved in and, and a solution of and was added to this and emulsified to obtain an injection.

Claims (2)

[Claims] 1. The following formula I (In the formula I, R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ₄ represents an oxygen atom, a sulfur atom or NH.) Compounds and pharmaceutically acceptable salts thereof. 2. The following formula I (In the formula I, R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R ₄ represents an oxygen atom, a sulfur atom or NH.) Anticancer agent containing the compound and a pharmaceutically acceptable salt thereof as an active ingredient. [0001]