JPH1081673A - Heterocyclic derivative, and carcinostatic containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound useful as a carcinostatic. SOLUTION: This new compound (salt) is shown by formula I (Het is a heterocycle such as substitutable thiazole or tetrazole cycle). e.g. 5-(3-amino-4- methoxyphenyl)-2-methyl-4-(3,4,5-trimethoxyphenyl)thiazole. The compound of formula I is obtained by oxidizing a compound of formula II with dimethyl sulfoxide and trifluoroacetic anhydride followed by reaction of the product with a thioamide derivative to form a thiazole derivative of formula III (X is a lower alkyl, amino, hydrazino, a halogen, lower alkoxy, etc.), which is then reduced with zinc in acetic acid. The compound of formula I is easy to synthesize, low in toxicity, and high in therapeutic effect.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a heterocyclic derivative and an anticancer agent containing such a compound as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION Combretastatins having cis-stilbene as a basic skeleton are known to exhibit strong mitotic inhibitory activity and have strong cytotoxicity, but the compounds are hardly soluble in water. For reasons, it has not been put to practical use as a pharmaceutical.

[0003]

DISCLOSURE OF THE INVENTION An object of the present invention is to find a combretastatin derivative which is easy to synthesize, has low toxicity and has a high therapeutic effect, and provides it as an anticancer agent.

[0004]

Means for Solving the Problems The present inventors have synthesized compounds in which two aromatic rings of various stilbene derivatives are fixed in a cis configuration, and intensively searched for compounds having anticancer activity. The compound represented by 1) was found to have remarkable anticancer activity in animal tests, and was found to have low toxicity, thereby completing the present invention. These compounds have not been synthesized so far and are completely novel compounds.

[0005]

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(In the formula, Het represents a heterocyclic ring.)

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION In the general formula (1), examples of the heterocyclic ring include a tetrazole ring and a thiazole ring.

When the heterocyclic ring in the general formula (1) is a thiazole ring, it may have a substituent, such as a lower alkyl group, an amino group, a monoalkylamino group, a dialkylamino group, a hydrazino group, Halogen and lower alkoxy groups.

Preferred compounds include the following compounds. 5- (3-amino-4-methoxyphenyl) -2-methyl-4- (3,4,5-trimethoxyphenyl) thiazole, 2-amino-5- (3-amino-4-methoxyphenyl) -4 -(3,4,5-trimethoxyphenyl) thiazole, 5- (3-amino-4
-Methoxyphenyl) -2-aminomethyl-4- (3,
4,5-trimethoxyphenyl) thiazole, 5- (3
-Amino-4-methoxyphenyl) -2-dimethylamino-4- (3,4,5-trimethoxyphenyl) thiazole, 5- (3-amino-4-methoxyphenyl) -2
-Hydrazino-4- (3,4,5-trimethoxyphenyl) thiazole, 5- (3-amino-4-methoxyphenyl) -2-methoxy-4- (3,4,5-trimethoxyphenyl) thiazole, 5- (3-Amino-4-methoxyphenyl) -2-chloro-4- (3,4,5-trimethoxyphenyl) thiazole.

5- (3-amino-4-methoxyphenyl) -1- (3,4,5-trimethoxyphenyl) tetrazole.

The compound of the present invention represented by the general formula (1) can be synthesized, for example, according to the following production route.

[0012]

Embedded image

(Wherein X is a lower alkyl group, an amino group,
It represents a substituent such as a monoalkylamino group, a dialkylamino group, a hydrazino group, a halogen, a lower alkoxy group and the like. )

[0014]

Embedded image

The compound represented by the formula (5) belonging to the compound of the present invention is, for example, E-2- (4-methoxy-3-nitrophenyl) -1- (3,4,5-trimethoxyphenyl) Ethene (the synthesis method is described in JP-A-7-228558)
With N-bromosuccinimide
Bromo-2- (4-methoxy-3-nitrophenyl)-
1- (3,4,5-trimethoxyphenyl) ethanol (2) is oxidized with dimethylsulfoxide and trifluoroacetic anhydride in dichloromethane at −78 ° C.
After 2-bromo-2- (4-methoxy-3-nitrophenyl) -1- (3,4,5-trimethoxyphenyl) ethanone (3), the mixture was added at room temperature in dimethylformamide in the presence of sodium carbonate. It can be obtained by reacting a thioamide derivative to synthesize a thiazole derivative (4), and reducing this with zinc in acetic acid.

The compound represented by the formula (9) belonging to the compound of the present invention can be obtained, for example, by reacting an acid chloride of 4-methoxy-3-nitrobenzoic acid with 3,4,5-trimethoxyaniline. The resulting (3-nitro-4-methoxyphenyl) -N- (3,4,5-trimethoxyphenyl) amide (6) is reacted with Lawesson's reagent in toluene at 60 ° C. to give (3-nitro-4-methoxyphenyl). Methoxyphenyl)
-N- (3,4,5-trimethoxyphenyl) -thioamide (7), followed by reaction with hydrazine hydrate, followed by reaction with trimethyl orthoformate to synthesize a tetrazole derivative (8). Medium, it can be obtained by reduction with zinc.

The heterocyclic derivative of the present invention produced by the above method can be easily separated and purified from the reaction mixture by conventional isolation and purification means, for example, extraction with a solvent, chromatography, crystallization and the like. it can.

In the present invention, when the above-mentioned heterocyclic derivative is used as an anticancer agent, it is administered orally or parenterally (intramuscularly, subcutaneously, intravenously, suppository, etc.). The dosage varies depending on the condition, but is usually 1 to 9000 mg per day in several divided doses per adult in general.

Further, when the heterocyclic derivative of the present invention is prepared as an oral preparation, an excipient and, if necessary, a binder, a disintegrant, a lubricant, a coloring agent, a flavoring agent and the like are added. Thereafter, tablets, coated tablets, granules, capsules and the like are prepared in a conventional manner.

The excipients include, for example, lactose, corn starch, sucrose, grapevine, sorbitol, crystalline cellulose, etc., and the binders include, for example, polyvinyl alcohol, polyvinyl ether, ethyl cellulose, methyl cellulose, gum arabic, tragacanth, gelatin, shellac, hydroxy Propyl cellulose, hydroxypropyl starch, polyvinylpyrrolidone, etc., as disintegrants, for example, starch, agar, gelatin powder, crystalline cellulose, calcium carbonate, sodium hydrogen carbonate, calcium citrate, dextran, pectin, etc., as lubricants Magnesium stearate, talc, polyethylene glycol, silica, hydrogenated vegetable oils, etc., are permitted to be added to pharmaceuticals as coloring agents,
As the flavoring agent, cocoa powder, peppermint brain, aromatic acid, peppermint oil, dragon brain, cinnamon powder and the like are used. In these tablets, the granules may be sugar-coated, gelatin-coated or any other appropriate coating as needed.

When preparing an injection, a pH adjuster, a buffer, a stabilizing agent, a preservative and the like are added as necessary, and a subcutaneous, intramuscular or intravenous injection is prepared by a conventional method.

The heterocyclic derivative of the present invention may be used, if necessary, with inorganic salts such as hydrochloric acid, sulfuric acid and phosphoric acid, and organic salts such as oxalic acid, fumaric acid, maleic acid, malic acid, citric acid, tartaric acid and glutamic acid. It can be a pharmaceutically acceptable acid addition salt.

[0023]

EXAMPLES The present invention will be described in detail with reference to the following Examples, but it should not be construed that the present invention is limited to these Examples.

Example 1 Synthesis of 2-amino-5- (3-amino-4-methoxyphenyl) -4- (3,4,5-trimethoxyphenyl) thiazole

Step 1 Synthesis of 2-bromo-2- (4-methoxy-3-nitrophenyl) -1- (3,4,5-trimethoxyphenyl) ethanol E-2- (4-methoxy-3-nitro) Phenyl) -1-
(3,4,5-trimethoxyphenyl) ethene 10.1
g was dissolved in 150 ml of dimethylsulfoxide + 10 ml of water, 7.5 g of N-bromosuccinimide was added, and the mixture was reacted at room temperature for 30 minutes. 500 ml of water was added, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure.
This reaction was performed four times, and a silica gel column (ethyl acetate-
Hexane, 1: 1 → 2: 1) and concentrated under reduced pressure.
The crude crystals were dissolved in ethyl acetate, hexane was slowly added thereto while heating, and the mixture was allowed to stand at room temperature.
1 mmol, 30% yield).

¹ H-NMR (CDCl ₃ ) 2.587 (1H, d, J = 3.2 Hz), 3.770 (6H, s), 3.814 (3H, s), 3.9
43 (3H, s), 5.070 (1H, d, J = 5.1Hz), 5.169 (1H, dd, J = 3.2H
z, 5.1Hz), 6.409 (2H, s), 6.959 (1H, d, 8.6Hz), 7.430 (1H,
(dd, J = 2.3Hz, 8.6Hz), 7.930 (1H, d, J = 2.3Hz)

Step 2 Synthesis of 2-bromo-2- (4-methoxy-3-nitrophenyl) -1- (3,4,5-trimethoxyphenyl) ethanone 5.5 ml of trifluoroacetic anhydride was added to dichloromethane 20
0 ml, cooled to −78 ° C., and added a solution of 5.0 ml of dimethyl sulfoxide + 40 ml of dichloromethane to 25 ml.
Dropped over minutes. After stirring at this temperature for 20 minutes, 2-
Bromo-2- (4-methoxy-3-nitrophenyl)-
A solution of 2.01 g of 1- (3,4,5-trimethoxyphenyl) ethanol in 25 ml of dichloromethane was added dropwise over 10 minutes, and the reaction was carried out at this temperature for 25 minutes. At this temperature 7.0 ml of triethylamine in 20 ml of dichloromethane
ml solution over 5 minutes and 2M hydrobromic acid 120m
1 was added and the temperature was raised to room temperature. The reaction solution was extracted with dichloromethane, and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. This was purified by a silica gel column (ethyl acetate-hexane, 2: 3) to obtain 2.14 g of the desired product.

¹ H-NMR (CDCl ₃ ) 3.919 (6H, s), 3.939 (3H, s), 3.985 (3H, s), 6.249 (1H,
s), 7.129 (1H, d, J = 8.9Hz), 7.273 (2H, s), 7.806 (1H, d-
d, J = 2.4Hz, 8.9Hz), 8.065 (1H, d, J = 2.4Hz)

Step 3 Synthesis of 2-amino-5- (4-methoxy-3-nitrophenyl) -4- (3,4,5-trimethoxyphenyl) thiazole 2-bromo-2- (4-methoxy-3) -Nitrophenyl) -1- (3,4,5-trimethoxyphenyl) ethanone 856 mg (1.94 mmol), thiourea 152
mg (2.00 mmol) of dimethylformamide 10
209 mg of sodium carbonate (1.97 m
mol) was added and reacted at room temperature for 2 hours. 50 ml of water
Was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, added thereto, allowed to cool, and further cooled to −20 ° C. to precipitate crystals. 615.4 mg of the desired product (1.47 m
mol, yield 76%).

[0030] _{1H-NMR (CDC l 3)} δ: 3.714 (6H, s), 3.848 (3
H, s), 3.946 (3H, s), 5.001 (2H, brs), 6.679 (2H, s), 6.98
4 (1H, d, J = 8.8Hz), 7.417 (1H, dd, J = 2.4Hz, 8.8Hz), 7.86
2 (1H, d, J = 2.4Hz)

Step 4 2-amino-5- (3-amino-4-methoxyphenyl)
Synthesis of 4- (3,4,5-trimethoxyphenyl) thiazole 12.2 g of activated zinc powder was suspended in 100 ml of acetic acid.
2-amino-5- at a rate such that the solution does not exceed 40 ° C.
(4-methoxy-3-nitrophenyl) -4- (3,
4,5-Trimethoxyphenyl) thiazole 585mg
A suspension of (1.40 mmol) in acetic acid (2.0 ml) + dichloromethane (4.0 ml) was added dropwise and reacted for 30 minutes.
300 ml of ethyl acetate was added and the mixture was filtered, and the filtrate was concentrated under reduced pressure. This was purified with a silica gel column (ethyl acetate-hexane 3: 2), concentrated, and then recrystallized from 100 ml of ethyl acetate + 200 ml of hexane to obtain 293.2 mg of the desired product
(0.757 mol, yield 54%, purity 97.7% -H
PLC).

[0032] _{1H-NMR (C D 3OD)} δ: 3.614 (6H, s), 3.706 (3
H, s), 3.811 (3H, s), 6.594 (1H, dd, 2.2Hz, 8.2Hz), 6.67
8 (1H, d, J = 2.2Hz), 6.719 (2H, s), 6.773 (1H, d, J = 8.2H
z); High-resolution mass spectrum (FAB): measured value 388.1313, calculated value 388.1331

Example 2 3- (3-amino-4-methoxyphenyl) -4- (3,
Synthesis of (4,5-trimethoxyphenyl) tetrazole

Step 1 (4-methoxyphenyl-3-nitro) -N- (3,
Synthesis method of 4,5-trimethoxyphenyl) amide 3.0 g of 4-methoxy-3-nitrobenzoic acid (0.01
5 mmol) was dissolved in 180 ml of dichloromethane, 3.3 ml (0.045 mmol) of thionyl chloride was added, and the mixture was stirred at room temperature for 12 hours. After concentrating the reaction solution, dichloromethane 150 ml, 3,4,5-trimethoxyaniline 3.0 g (0.017 mmol), pyridine 5 ml
Was added and reacted at room temperature for 2 hours. Then, saturated aqueous sodium hydrogen carbonate was added, and the mixture was extracted three times with dichloromethane. After drying over anhydrous sodium sulfate, the mixture was concentrated and purified by silica gel column chromatography (5% methanol-dichloromethane). After concentration, the residue was dissolved in dichloromethane and recrystallized by adding ethyl acetate to obtain 4.2 g of the desired product (yield: 77.
3%).

[0035] _{1H-NMR (CDC l 3)} δ: 8.35 (1H, d, J = 2.4),
8.15 (1H, dd, J = 2.4,8.7), 7.92 (1H, brs), 7.19 (1H, d, J =
8.7), 6.96 (2H, s), 4.04 (3H, s), 3.87 (6H, s), 3.84 (3H, s
s); molecular weight _{C1 7 H1 8 N 2 O 7} = 362; ESI mass spectrum: 363 (MH
+); Yellow crystal Melting point 168-169 ℃

Step 2 N- (4-methoxyphenyl-3-nitro)-(3,4,5
Synthesis of (trimethoxyphenyl) thioamide (4-methoxyphenyl-3-nitro) -N- (3,4,
2.0 g of 5-trimethoxyphenyl) amide was dissolved in 100 ml of toluene, 2.68 g of Lawesson's reagent was added, and the mixture was reacted at 100 ° C. for 3 hours. After cooling to room temperature and concentrating, ethyl acetate and hexane were added in that order for crystallization to obtain 1.3 g (3.4 mmol, 61.8%) of the desired product.

[0037] _{1H-NMR (CDC l 3)} δ: 9.03 (1H, s), 8.19 (1
H, s), 8.05 (1H, brs), 7.15 (1H, d, J = 9.0), 7.04 (2H,
s), 4.01 (3H, s ), 3.91 (6H, s), 3.89 (3H, s); molecular weight _{C1 7 H1 8 N 2 O 6} S 1 = 378; ESI mass spectrum: 379 (MH
+); Yellow crystal Melting point 199-200 ℃

Step 3 3- (4-methoxyphenyl-3-nitro) -4- (3,
Synthesis of 4,5-trimethoxyphenyl) tetrazole (4-methoxyphenyl-3-nitro) -N- (3,4,5
-Trimethoxyphenyl) thioamide 504 mg (1.
33 mmol) was dissolved in 25 ml of ethanol + 25 ml of dichloromethane, 5 ml of hydrazine hydrate was added, and the mixture was reacted at room temperature for 2 hours. Next, the reaction solution was concentrated, the concentrate was dissolved in 25 ml of acetic acid, 300 mg of sodium nitrite was added, and the mixture was reacted at room temperature for 4 hours. After neutralization with 2N sodium hydroxide, saturated brine was added, and then dichloromethane was added. Extracted. After drying over anhydrous sodium sulfate, the mixture was concentrated, crystallized from ethyl acetate, and 483 mg of the desired product (1.28 mm
ol, 80.0%).

[0039] _{1H-NMR (CDC l 3)} δ: 8.02 (1H, d, J = 2.7),
7.63 (1H, dd, J = 2.7,9.0), 7.25 (1H, d, J = 9.0), 6.99 (2
H, s), 4.06 (3H , s), 3.91 (3H, s), 3.75 (6H, s); molecular weight _{C1 7 H1 7 N 5 O 6} = 387; ESI mass spectrum: 388 (MH +)
; White crystals, melting point 197-198 ° C

Step 4 3- (3-amino-4-methoxyphenyl) -4- (3,
Synthesis of 4,5-trimethoxyphenyl) tetrazole 3- (4-methoxyphenyl-3-nitro) -4- (3,
4,5-trimethoxyphenyl) tetrazole 400m
g in 20 ml of acetic acid + 10 ml of dichloromethane,
5 g of activated zinc was added, and the mixture was stirred at room temperature for 2 hours. After filtration and concentration, silica gel column chromatography (5%
Methanol-dichloromethane) to give the desired product 257
mg (0.75 mmol, 70.1%).

[0041] _{1H-NMR (CDC l 3)} 6.89 (2H, s), 6.85 (1H, d, J = 8.1), 6.73 (1H, d, J = 2.4), 6.
71 (1H, dd, J = 2.4,8.1), 3.92 (3H, s), 3.88 (3H, s), 3.71 (6
H, s) molecular weight _{C1 7 H1 9 N 5 O 4} = 357; ESI mass spectrum 358 (MH
+); White crystal Melting point 147-148 ° C High-resolution mass spectrum (FAB): (MH +) calculated 358.151
5 Measured value 358.1515

Example 3 Evaluation of Cytotoxicity Mouse colon 26 colon cancer cells were used as cancer cells, and a PRMI-1640 medium containing 10% fetal bovine serum was used for culture. 5 × 103/50 on a 96-well microplate
After inoculation at 50 μl / well, the test compound aqueous solution of each concentration was added at 50 μl / well, and cultured at 37 ° C. for 3 days.
The number of viable cells was measured by the T method, and a dose response curve was prepared. From this it was calculated 50% growth inhibitory concentration by the test compound (IC ₅ 0). The IC ₅ 0 values of each compound shown in Table 1. In addition, the dose at which an acute death occurred immediately after administration of the drug intravenously is shown in Table 1 as a toxicity manifestation dose.

Example 4 Drug Efficacy Test in Mice Colon 26 subcutaneously passaged in mice was crushed with scissors and transplanted subcutaneously in mice with a trocar. One week later, the tumor was measured with a caliper, and the tumor volume was calculated to perform grouping (n = 5 in each group). After the test compound was dissolved in dimethyl sulfoxide, it was diluted with a 5% Tween 80-physiological saline solution, and 0.2 ml of the solution was administered by intravenous injection or subcutaneous injection. Drug administration on day 7 after transplantation, 1
The test was performed once a day on the 1st and 15th days. Tumor volume was measured 21 days after transplantation, and tumor volume and tumor growth inhibition rate were measured.
(I.R.) was calculated by Equations 1 and 2, respectively.

[0044]

(Equation 1)

[0045]

(Equation 2)

[0046]

【table 1】

[0047]

The heterocyclic derivative of the present invention has excellent anticancer activity and is extremely useful in the pharmaceutical industry.

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[Procedure amendment]

[Submission date] November 26, 1996

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0016

[Correction method] Change

[Correction contents]

The compound represented by the formula (9) belonging to the compound of the present invention can be obtained, for example, by reacting an acid chloride of 4-methoxy-3-nitrobenzoic acid with 3,4,5-trimethoxyaniline. Obtained ( 4-methoxy-3-nito)
B phenyl)-N-(3, 4,5-trimethoxyphenyl) in toluene amide (6) is reacted with Lawesson's's reagent 60 ° C. (4-methoxy-3-nitrophenyl)
-N- (3,4,5-trimethoxyphenyl) -thioamide (7), followed by reaction with hydrazine hydrate, followed by reaction with trimethyl orthoformate to synthesize a tetrazole derivative (8). Medium, it can be obtained by reduction with zinc.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0026

[Correction method] Change

[Correction contents]

¹ H-NMR (CDCl ₃ ) δ : 2.587 (1H,
d, J = 3.2 Hz), 3.770 (6H, s),
3.814 (3H, s), 3.943 (3H, s),
5.070 (1H, d, J = 5.1 Hz), 5.1
69 (1H, dd, J = 3.2 Hz, 5.1 Hz),
6.409 (2H, s), 6.959 (1H, d,
8.6 Hz), 7.430 (1H, dd, J = 2.
3Hz, 8.6Hz), 7.930 (1H, d, J =
2.3Hz)

[Procedure amendment 3]

[Document name to be amended] Statement

[Correction target item name] 0028

[Correction method] Change

[Correction contents]

¹ H-NMR (CDCl ₃ ) δ : 3.9
19 (6H, s), 3.939 (3H, s),
985 (3H, s), 6.249 (1H, s),
7.129 (1H, d, J = 8.9 Hz), 7.27
3 (2H, s), 7.806 (1H, dd, J =
2.4 Hz, 8.9 Hz), 8.065 (1H, d,
J = 2.4Hz)

[Procedure amendment 4]

[Document name to be amended] Statement

[Correction target item name] 0030

[Correction method] Change

[Correction contents]

¹ H-NMR (CDCl ₃ ) δ: 3.7
2. 14 (6H, s), 3.848 (3H, s),
946 (3H, s), 5.001 (2H, brs),
6.679 (2H, s), 6.984 (1H, d, J
= 8.8 Hz), 7.417 (1H, dd, J =
2.4Hz, 8.8Hz), 7.862 (1H, d,
J = 2.4Hz)

[Procedure amendment 5]

[Document name to be amended] Statement

[Correction target item name] 0032

[Correction method] Change

[Correction contents]

¹ H-NMR (CD ₃ OD) δ: 3.614
(6H, s), 3.706 (3H, s), 3.811 (3H, s), 6.594 (1H, dd, 2.2
Hz, 8.2Hz), 6.678 (1H, d, J = 2.2Hz), 6.719 (2H, s), 6.773
(1H, d, J = 8.2Hz); High resolution mass spectrum (FAB): measured value 388.1313, calculated value 388.1331

[Procedure amendment 6]

[Document name to be amended] Statement

[Correction target item name] 0034

[Correction method] Change

[Correction contents]

Step 1 ( 4-methoxy-3-nitrophenyl ) -N- (3,
Synthesis method of 4,5-trimethoxyphenyl) amide 3.0 g of 4-methoxy-3-nitrobenzoic acid (0.01
5 mmol) was dissolved in 180 ml of dichloromethane, 3.3 ml (0.045 mmol) of thionyl chloride was added, and the mixture was stirred at room temperature for 12 hours. After concentrating the reaction solution, dichloromethane 150 ml, 3,4,5-trimethoxyaniline 3.0 g (0.017 mmol), pyridine 5 ml
Was added and reacted at room temperature for 2 hours. Then, saturated aqueous sodium hydrogen carbonate was added, and the mixture was extracted three times with dichloromethane. After drying over anhydrous sodium sulfate, the mixture was concentrated and purified by silica gel column chromatography (5% methanol-dichloromethane). After concentration, the residue was dissolved in dichloromethane and recrystallized by adding ethyl acetate to obtain 4.2 g of the desired product (yield: 77.
3%).

[Procedure amendment 7]

[Document name to be amended] Statement

[Correction target item name] 0035

[Correction method] Change

[Correction contents]

¹ H-NMR (CDCl ₃ ) δ: 8.35 (1H, d, J = 2.4), 8.
15 (1H, dd, J = 2.4,8.7), 7.92 (1H, brs), 7.19 (1H, d, J = 8.
7), 6.96 (2H, s), 4.04 (3H, s), 3.87 (6H, s), 3.84 (3H, s
s); molecular weight C ₁₇ H ₁₈ N ₂ O ₇ = 362; ESI mass spectrum: 363 (MH
+); Yellow crystal Melting point 168-169 ℃

[Procedure amendment 8]

[Document name to be amended] Statement

[Correction target item name] 0036

[Correction method] Change

[Correction contents]

Step 2 N- ( 4-methoxy-3-nitrophenyl )-( 3,4,5
Synthesis of (trimethoxyphenyl) thioamide ( 4-methoxy-3-nitrophenyl ) -N- (3,4,
2.0 g of 5-trimethoxyphenyl) amide was dissolved in 100 ml of toluene, 2.68 g of Lawesson's reagent was added, and the mixture was reacted at 100 ° C. for 3 hours. After cooling to room temperature and concentrating, ethyl acetate and hexane were added in that order for crystallization to obtain 1.3 g (3.4 mmol, 61.8%) of the desired product.

[Procedure amendment 9]

[Document name to be amended] Statement

[Correction target item name] 0037

[Correction method] Change

[Correction contents]

¹ H-NMR (CDCl ₃ ) δ: 9.03 (1H, s), 8.19 (1
H, s), 8.05 (1H, brs), 7.15 (1H, d, J = 9.0), 7.04 (2H,
s), 4.01 (3H, s), 3.91 (6H, s), 3.89 (3H, s); molecular weight C ₁₇ H ₁₈ N ₂ O ₆ S ₁ = 378; ESI mass spectrum: 379 (MH
+); Yellow crystal Melting point 199-200 ℃

[Procedure amendment 10]

[Document name to be amended] Statement

[Correction target item name] 0038

[Correction method] Change

[Correction contents]

Step 3 3- ( 4-methoxy-3-nitrophenyl ) -4- (3,
Synthesis of 4,5-trimethoxyphenyl) tetrazole ( 4-methoxy-3-nitrophenyl ) -N- (3,4,5
-Trimethoxyphenyl) thioamide 504 mg (1.
33 mmol) was dissolved in 25 ml of ethanol + 25 ml of dichloromethane, 5 ml of hydrazine hydrate was added, and the mixture was reacted at room temperature for 2 hours. Next, the reaction solution was concentrated, the concentrate was dissolved in 25 ml of acetic acid, 300 mg of sodium nitrite was added, and the mixture was reacted at room temperature for 4 hours. After neutralization with 2N sodium hydroxide, saturated brine was added, and then dichloromethane was added. Extracted. After drying over anhydrous sodium sulfate, the mixture was concentrated, crystallized from ethyl acetate, and 483 mg of the desired product (1.28 mm
ol, 80.0%).

[Procedure amendment 11]

[Document name to be amended] Statement

[Correction target item name] 0039

[Correction method] Change

[Correction contents]

¹ H-NMR (CDCl ₃ ) δ: 8.02 (1H, d, J = 2.7),
7.63 (1H, dd, J = 2.7,9.0), 7.25 (1H, d, J = 9.0), 6.99 (2
H, s), 4.06 (3H , s), 3.91 (3H, s), 3.75 (6H, s); molecular weight _{C 17 H 17 N 5 O 6} = 387; ESI mass spectrum: 388 (MH +)
; White crystals, melting point 197-198 ° C

[Procedure amendment 12]

[Document name to be amended] Statement

[Correction target item name] 0040

[Correction method] Change

[Correction contents]

Step 4 3- (3-amino-4-methoxyphenyl) -4- (3,
Synthesis of 4,5-trimethoxyphenyl) tetrazole 3- ( 4-methoxy-3-nitrophenyl ) -4- (3,
4,5-trimethoxyphenyl) tetrazole 400m
g in 20 ml of acetic acid + 10 ml of dichloromethane,
5 g of activated zinc was added, and the mixture was stirred at room temperature for 2 hours. After filtration and concentration, silica gel column chromatography (5%
Methanol-dichloromethane) to give the desired product 257
mg (0.75 mmol, 70.1%).

[Procedure amendment 13]

[Document name to be amended] Statement

[Correction target item name] 0041

[Correction method] Change

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¹ H-NMR (CDCl ₃ ) δ : 6.89 (2H, s), 6.85 (1
H, d, J = 8.1), 6.73 (1H, d, J = 2.4), 6.71 (1H, dd, J = 2.4,8.
1), 3.92 (3H, s ), 3.88 (3H, s), 3.71 (6H, s) Molecular weight _{C 17 H 19 N 5 O 4} = 357; ESI mass spectrum 358 (MH
+); White crystal Melting point 147-148 ° C High-resolution mass spectrum (FAB): (MH +) calculated 358.151
5 Actual value 358.1515

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Example 3 Evaluation of Cytotoxicity Mouse colon 26 colon cancer cells were used as cancer cells, and PRMI-164 containing 10% fetal bovine serum was used for culture.
0 medium was used. 5 × 10 on 96-well microplate
^The cells were seeded at ³ cells / 50 μl / well, and the test compound aqueous solution of each concentration was added in an amount of 50 μl at a time, and the cells were cultured at 37 ° C. for 3 days. The number of viable cells was measured by the MTT method, and a dose response curve was prepared. From this, the 50% growth inhibitory concentration ( IC ₅₀ ) of the test compound was calculated. The IC ₅₀ values of each compound are shown in Table 1. In addition, the dose at which an acute death occurred immediately after administration of the drug intravenously is shown in Table 1 as a toxicity manifestation dose.

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【table 1】

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Naoshi Tsuji 1-1, Suzukicho, Kawasaki-ku, Kawasaki-shi, Kanagawa Prefecture Ajinomoto Co., Inc. 1. Ajinomoto Co., Ltd. Central Research Laboratory (72) Inventor Yukio Nihei 1-1, Suzukicho, Kawasaki-ku, Kawasaki-shi, Kanagawa Prefecture Ajinomoto Co. Central Research Laboratory

Claims (3)

[Claims] 1. A heterocyclic derivative represented by the following general formula (1). Embedded image (In the formula, Het represents a heterocyclic ring.) 2. The heterocyclic derivative according to claim 1, wherein the heterocyclic ring is thiazole or tetrazole which may have a substituent. 3. A carcinostatic agent comprising the heterocyclic derivative according to claim 1 or a pharmaceutically acceptable salt thereof.