JPS62273979A - 1,5-substituted-1h-pyrazolo(3,4-b)pyrazine derivative and antitumor agent containing said compound - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I [R1 is H, OH, halogen, hydrazino, OR' or NHR' (R' is alkyl, benzyl or phenyl or phenyl which may be substituted); R2 is vinyl or CH2CH2X (X is halogen)]. EXAMPLE:1-(2-Chloroethyl)-1H-pyrazolo[3,4-b]pyrazine. USE:An antitumor agent, having relatively low toxicity and effective even against solid cancer. PREPARATION:For example, 5-amino-1-(2-hydroxyethyl)-pyrazole expressed by formula II is reacted with isoamyl nitrite in the presence of hydrochloric acid in ethanol to give a compound expressed by formula III, which is then catalytically hydrolyzed using Pd-C, etc. The catalyst is then removed and the resultant product is reacted with glyoxal to afford a compound expressed by formula IV, which is reacted with thionil chloride in the presence of pyridine, etc., to halogenate the hydroxyethyl group and give the aimed compound expressed by formula I (R1 is H; R2 is CH2CH2X).

Description

[Detailed description of the invention] industrial misfortune The present invention is based on the following formula (1) [However, R is a hydrogen atom, a hydroxyl group, or a halogen atom.

hydrazino group, -0-R' group or -NH-R' group (where R' is an alkyl group, benzyl group, phenyl group, or one or more hydrogen atoms of the phenyl group is a halogen atom, an alkyl group, or an alkoxy group) R2 represents a vinyl group or a -CH2CH,X group (herein or a halogen atom). The present invention relates to novel 1,5-substituted fi-IH-pyrazolo[3,4-bl pyrazine derivatives] and antitumor agents containing these compounds as active ingredients.

Problems that future techniques are trying to solve
Various substances have been proposed as having antitumor effects,
Although it is known that certain IH-pyrazolo[3,4-bl pyrazine derivatives have antitumor effects, there are even more excellent anti-Ii1. There is a desire for a substance that has tumor activity, low toxicity, and can be effectively used as an antitumor agent.

I- In view of the above circumstances, the present invention has relatively low toxicity.

As a result of extensive research into substances with excellent antitumor activity, we discovered that 5-chloro-1-(2-hydroxyethyl)-
A 5-substituted-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine derivative was synthesized by reacting 1H-pyrazolo[3,4-bl pyrazine with hydrazine or an amino compound. By reacting thionyl halide, a new compound 5-substituted-1-(2-halogenoethyl)-1H-pyrazolo[3°4-b]pyrazine derivative can be obtained, and by further hydrolyzing this, or 5-chloro-1-(2-halogenoethyl)-LH-
By reacting pyrazolo C3,4-bl pyrazine with alkoxide, various new compounds 1.5-1i
In addition to discovering that converted-IH-pyrazolo[3゜4-b]pyrazine derivatives can be obtained, these new compounds all have excellent antitumor effects and relatively low toxicity, making them useful as antitumor agents. The present invention was based on the discovery that it can be used effectively.

Therefore, the present invention is based on the following formula (1) [where R□ is a hydrogen atom, a hydroxyl group, a halogen atom, a hydrazino group, a -0-R' group, or a -NH-R' group (where R' is an alkyl group] , a benzyl group, a phenyl group, or a phenyl group in which one or more hydrogen atoms of the phenyl group are substituted with a halogen atom, an alkyl group, or an alkoxy group)
, R2 is a vinyl group or -CH, CH, X group (here,
or a halogen atom). ] New 1 indicated by
, 5-substituted-IH-pyrazolo[3,4-bl pyrazine derivative and an antitumor agent containing this new compound as an active ingredient.

The present invention will be explained in more detail below.

The novel compound 1 according to the present invention, the 5-@substituted-IH-pyrazolo[3,4-b]pyrazine derivative, has the formula (1) described above, that is, [where R□ is a hydrogen atom, a hydroxyl group, a halogen atom, a hydrazine atom] group, -0-R' group or -N) (-R' group (where R' is an alkyl group, benzyl group, phenyl group, or one or more hydrogen atoms of the phenyl group is a halogen atom, an alkyl group, or an alkoxy group) R2 represents a vinyl group or a -CH2CH,x group (wherein, X is a halogen atom).

Here, the halogen atom is CR, Br, etc., and the alkyl group is CnI (2rl+, (however, n=1 to
8), etc., and examples of the alkoxy group include ○CnH, n, (where n=1 to 8), and the like.

Among the compounds represented by the above general formula (1).

5-amino-1-( represented by the following formula (1a) CH2CH,
2-halogenoethyl)-1H-pyrazolo[3,4-b]
The pyrazine dicenter and the 5-amino-1-vinyl-IH-pyrazolo[3,4-bl pyrazine derivative represented by the following formula (1b) can be synthesized, for example, by the following method (see reaction formula A below) .

That is, 5-amino-1-(2-hydroxyethyl)-pyrazole (formula (2) below) is reacted with isoamyl nitrite in ethanol in the presence of hydrochloric acid to form 5-amino-1-(2-hydroxyethyl)-pyrazole (formula (2) below).
-Hydroxyethyl)-4-nitroso-pyrazole hydrochloride (formula (3) below) was obtained, which was catalytically reduced using palladium-carbon, etc., the catalyst was removed, and then it was dissolved in water or methanol. React with glyoxylic acid to synthesize 5-hydroxy-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (formula (4) below),
This was treated with acetic anhydride to form 5-acetyloxy-1-
(2-acetyloxyethyl)-1H-pyrazolo[3,
4-bl pyrazine (formula (5) below) is obtained. Next, this is hydrolyzed using a partial hydrolyzing agent such as C, H, NH, etc. to obtain 5-hydroxy-1-(2-acetyloxy). ethyl)-LH-pyrazolo[3,4-bl pyrazine (formula (6) below) was obtained, and 5-chloro-1-(2-acetyloxyethyl)-1H-pyrazolo[ 3,4-bl pyrazine (the following formula (
7)) and further hydrolyzed with alkali to give 5-chloro-
1-(2-hydroxyethyl)-1H-pyrazolo[3,
4-bl pyrazine (formula (8) below) is reacted with an amino compound to form a 5-amino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine derivative (formula (8) below). After synthesizing 9)), 5-amino-1-(2-halogenoethyl)-1H-pyrazolo[3,4 -bl pyrazine derivative (formula 1a) is obtained. This was reacted with an alkali to form 5-amino-1-vinyl-IH-pyrazolo [3゜4-
b] Pyrazine derivative (21b) is obtained.

■ CH3CO,OH' CH3CO,0HCH3C
O,0H (5)CH,CH,0COCH.

CM, CH, ○cOcH.

CH3CO,0COCH.

CH3CO,0H CH,CH20H CH2CH2X (1a) (1b) Furthermore, among the compounds represented by the above general formula (1), 5-hydroxy-1-(2-halogenoethyl represented by the following formula (1c) CH2CH,X )-1H-pyrazolo[3,4-bl pyrazine and the following formula (1d) CH=CH.

5-hydroxy-1-vinyl-IH-pyrazolo[3,4-bl pyrazine represented by can be synthesized, for example, by the following method (see reaction formula B below).

That is, 5-hydroxy-1-(2-hydroxyethyl)-LH-pyrazolo[3,4-bl pyrazine of the above formula (4) is reacted with thionyl chloride in the presence of pyridine to halogenate the hydroxyethyl group, 5-hydroxy-
1-(2-halogenoethyl)-1H-pyrazolo[3,4
-b] pyrazine (formula 1c) is obtained. Furthermore, by reacting this with an alkali, 5-hydroxy-1-vinyl-IH-pyrazolo[3,4-b]pyrazine (formula 1% formula %). (1e) 5-chloro-1-(2-halogenoethyl)- represented by CH, CH,
1H-pyrazolo[3,4-b]pyrazine, the following formula (1f
)CH=CH.

5-chloro-1-vinyl-IH-pyrazolo [
3,4-b copyrazine and 5-hydrazino-1-vinyl-IH-pyrazolo[3,4-bl pyrazine represented by the following formula (1g) CH=CH2] can be synthesized, for example, by the following method (the following reaction Formula C).

That is, the hydroxyethyl group is halogenated by reacting 5-chloro-1-(2-hydroxyethyl)-LH-pyrazolo[3,4-bl pyrazine of the above formula (8) with thionyl chloride in the presence of pyridine. or above (IC)
5-hydroxy-1-(2-halogenoethyl)-1 of the formula
Synthesize 5-chloro-1-(2-halogenoethyl)-1H-pyrazolo[3°4-b]pyrazine (Formula 1e) by reacting H-pyrazolo[3,4-bl pyrazine with phosphorus oxychloride. By reacting this with t-butoxide, 5-chloro-1-vinyl-IH-pyrazolo[3,
4-b] pyrazine (formula 1f) is obtained by reacting it with hydrazine hydrate in ethanol,
(Ig) formula % formula % [3 4-b] pyrazine is obtained.

(1c) CH=CH.

(1f) CH=CH.

(1g) Also, among the compounds represented by the above general formula (1), 1-(2-halogenoethyl)-1H-pyrazolo[3,4-bl pyrazine] represented by the following formula (1h) CH,CH,X and 1-vinyl-IH-pyrazolo[3,4-bl
Pyrazine can be synthesized, for example, by the following method (see the reaction formula below).

That is, after catalytic reduction of 5-amino-1-(2-hydroxyethyl)-4-nitroso-pyrazole hydrochloride of the following formula (3) using palladium-carbon etc., the catalyst was removed and 1
Next, this is reacted with glyoxal in water or methanol to synthesize 1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (formula 10 below), and this is added to thionyl chloride in the presence of pyridine. By reacting, etc.
By halogenating the hydroxyethyl group, 1-
(2-halogenoethyl)-1H-pyrazolo[3,4-b
The l pyrazine derivative (formula 1h) is obtained. By reacting this with an alkali, 1-vinyl-IH-pyrazolo[3
, 4-b] pyrazine derivative (formula 1i).

(1o) (1h) CHa CHa ,4-b] pyrazine derivative is 1, for example, the above (1
d) 5-chloro-(2-halogenoethyl)-10- of the formula
Pyrazolo[3,4-bl pyrazine or the above formula (1h)%
Formula % 11? can be obtained by reacting pyrazine with alkoxide (R″-ON a ) (see reaction formulas E and E′ below)
.

(ld) (lj) Above (1)
The compound of the formula ) has an excellent antitumor effect and suppresses the proliferation of cancerous cells, so it is effectively used as an antitumor agent.

In addition, when the compound of formula (1) is used as an anti-Eft tumor agent, the halogen atom in formula (1) is CQ or Br, the alkyl group is CnH2n+ (however, n = 1 to 8).
Those in which the alkoxy group is OCnH-n+z (where n=1 to 8) are preferred.

The antitumor agent according to the present invention contains a compound of formula (1) as an active ingredient, and these compounds of formula (1) can be administered by intravenous injection or subcutaneous injection alone or in combination with other pharmaceutical ingredients if necessary. It can be administered by methods such as intramuscular injection, oral administration, or rectal administration using a waste drug. The dosage varies depending on the route of administration, the number of times of administration, etc., and can be varied over a wide range depending on the severity of the symptoms, etc., but in general, the therapeutically effective dosage is about 1 to 1 to 1 kg of the active ingredient of the present invention per 1 kg of body weight per day. It is 50■.

The antitumor agent according to the present invention can be prepared for administration by combining an effective amount of the compound represented by general formula (1) with an appropriate amount of non-toxic carrier and using any conventional formulation method. That is, when prepared for oral administration, it is prepared into soft capsules, hard capsules, tablets, granules, fine granules, powders, active ingredient sustained release agents, solutions, suspensions, etc., and when administered parenterally. is prepared into injections, drips, suppositories, etc. in this case,
When formulating, non-toxic carriers such as alcohols, esters, polyethylene glycol H8, sorbitan fatty acid esters, surfactants such as sulfated fatty alcohols, gum arabic, gelatin, sorbitol, gum tragacanth, polyvinylpyrrolidone, etc. are combined. agents, sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium aluminate metasilicate, synthetic aluminum silicate, calcium carbonate, sodium bicarbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose, etc. excipients,
Magnesium stearate, talc, lubricants such as hydrogenated oil, salt, saccharin, orange oil, licorice extract, citric acid, glucose, menthol, eucalyptus oil, flavoring agents such as malic acid, deodorants, coconut oil, olive oil, Suspending agents such as sesame oil, peanut oil, calcium lactate, safflower oil, soybean phospholipids, wetting agents, cellulose acetate phthalate (C
Cellulose such as AP), carbohydrate derivatives such as saccharides, acrylic acid copolymers such as methyl acrylate/methacrylic acid copolymer, methyl methacrylate/methacrylic acid copolymer, dibasic acid monoesters, etc. It is prepared by a conventional method using a polyvinyl derivative of , other film-forming agents, coating aids, and other components, and then used. In addition, when preparing preparations for mucosal application, especially waste preparations, cacao butter, lauric fat, polyethylene glycol, glycerogelatin, sodium stearate, or a mixture thereof is used as a base. Furthermore, injections are prepared by conventional methods, but when suspending or emulsifying in distilled water for injection is used, suspending agents such as sorbitol syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, and aluminum stearate gel are used. etc. can be used, and sorbitan monooleate, polyoxyethylene hydrogenated castor oil, lecithin, etc. can be used as emulsifiers.

The novel compound of the present invention has antitumor activity and is therefore useful as an antitumor agent.An antitumor agent containing this new compound as an active ingredient has such excellent antitumor activity. ,
It has relatively low toxicity and is effective against solid cancers.

Next, a production example of the compound (1) of the present invention will be shown.

[Production Example 1] 20 mM of ethanol containing 3.65 g of hydrogen chloride gas was cooled to 0 to 5°C, and 5-amino-1 of the formula (2) was added to it with stirring while keeping the reaction temperature below 5°C. -(2-Hydroxyethyl)-pyrazole is dissolved, and 12 g of isoamyl nitrite is added dropwise thereto. Leave it at room temperature overnight and filter the precipitated crystals.

12.5 g (yield 65%) of 5-amino-1-(2-hydroxyethyl)-4-nitroso-pyrazole hydrochloride (compound 2 of formula (3) decomposition temperature 152-154°C) is obtained.

The above 5-amino-1-(2-hydroxyethyl)-4-
12.5 g of nitroso-pyrazole hydrochloride was dissolved in methanol 2
After dissolving in 00aQ and adding 5 g of palladium-carbon to perform catalytic reduction, the catalyst was removed.

This is reacted with 10.1 g of a 40% glyoxal aqueous solution. Distill off the methanol and add ice water.

After neutralization, extraction with chloroform yielded 1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (
Compound of formula (10), melting point 87-88°C) 7.58
get g.

The above 1-(2-hydroxyethyl)-LH-pyrazolo[
3,4-b] pyrazine 7, sag in chloroform 50 m
Dissolve in Q and add 5.5 g of pyridine and 8 thionyl chloride.
．． Add 27 g and reflux for 1 hour. After the reaction, the chloroform layer was poured into ice water and washed with water to obtain 1-(2-chloroethyl)-1H-pyrazolo[3,4-b]pyrazine (compound of formula (1h) above, VA point 64-65°C )6.74g
(yield 80%).

[Production Example 2] 12.5 g of the compound of formula (3) above was added to 200 m of methanol.
After catalytic reduction by adding 5 g of palladium-carbon to the solution, the catalyst was removed, and 9.74 g of a 50% glyoxylic acid aqueous solution was reacted with the solution, and the precipitated crystals were collected by filtration. -Hydroxy-1-(2-hydroxyethyl)-1H-pyrazolo[3°4-b]pyrazine (compound of formula (4), melting point 259-260°C) 7.0 g (yield 60%) is obtained.

The above 5-hydroxy-1-(2-hydroxyethyl)-
7.0 g of 1H-pyrazolo[3,4-b]pyrazine was dissolved in 50 mQ of chloroform, and 5.5 g of pyridine,
Add 8.27 g of thionyl chloride and reflux for 1 hour. After the reaction, the chloroform layer was poured into ice water and washed with water to give 6.56 g (yield) of 5-hydroxy-1-(2-chloroethyl)-1H-pyrazolo[3,4-b]pyrazine (formula %). 85%).

[Production Example 3] The above 5-hydroxy-1-(2-chloroethyl)-1H
5-chloro-1-(2-chloroethyl)-1H-pyrazolo[3,4-bl pyrazine (formula (1e) Compound, melting point 79-80°C) 3.5
6 g (82% yield) are obtained.

[Production Examples 4 to 9] In Production Example 4, the above 5-hydroxy-1-(2-
180.0 g of hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine was reacted with 500 grams of acetic anhydride to give 5
-acetyloxy-1-(2-acetyloxyethyl)
-LH-pyrazolo[3,4-b copyrazine (formula (5)
) compound, melting point 85-86℃) 260.0g (yield 9
8%).

The above 5-acetyloxy-1-(2-acetyloxyethyl)-1H-pyrazolo[3,4-bl pyrazine 264
．． 0g was dissolved in ether 500-, and 12.0.0g of hexylamine was reacted with it to form 5-hydroxy-1-
(2-acetyloxyethyl)-1H-pyrazolo[3,
4-b] Pyrazine (compound of formula (6) above, melting point 173
~174°C) 211.0 g (yield: $95%) was obtained.

The above 5-hydroxy-1-(2-acetyloxyethyl)-LH-pyrazolo[3,4-b cobylazine 222.0
g with 500 mQ of phosphorus oxychloride to form 5-chloro-1-(2-acetyloxyethyl)-111-pyrazolo[3,4-b]pyrazine (the compound of formula (7) above,
(melting point 52-53°C) 190.0 g (yield 79%) is obtained.

The above 5-chloro-1-(2-acetyloxyethyl)-
10% aqueous sodium hydroxide solution was added to 120.3 g of 1H-pyrano[3,4-b]pyrazine and hydrolyzed,
Obtain 90.4 g (yield 91%) of 5-chloro-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (compound of formula (8) above, melting point 100-102°C) .

The above 5-chloro-1-(2-hydroxyethyl)-1H
- 3.97 g of pyrazolo[3,4-b]pyrazine and 10 mQ of hexylamine were reacted in a sealed tube, excess amine was removed, and 5-hexylamino-1-(2-hydroxyethyl)-1H-pyrazolo[ 3,4-bl pyrazine (
4.48 g (yield: 85%) of the compound of formula (9) is obtained.

2.63 g of the above 5-hexylamino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine
Dissolved in chloroform 50-, add pyridine 1.7
g, 2.5 g of thionyl chloride were added, and the mixture was refluxed for 1 hour. After the reaction, the chloroform layer was poured into ice water and washed with water.
-hexylamino-1-(2-chloroethyl)-1H-
2.22 g (yield: 84%) of pyrazolo[3,4-b]pyrazine (compound of formula (1a) above, melting point 80-81°C) is obtained.

Production Example 5 uses benzylamine instead of hexylamine, and Production Example 6 is operated in the same manner as Production Example 4 using aniline.

In Production Example 7, 3.97 g of the above 5-chloro-1-(2-hydroxyethyl)-LH-pyrazolo[3,4-bl pyrazine and 4.0 g of parachloroaniline were reacted in a sealed tube to remove excess Except for amines.

5-(4-chlorophenylamino)-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (1) "Compound of formula (9), melting point 187-188°C)"
Obtain 5.33 g (92% yield).

2.90 g of the above 5-(4-chlorophenylamino)-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine was dissolved in 50-chloroform, and 1.7 g of pyridine and 2.2 g of thionyl chloride were dissolved in 50-chloroform. Add .5 g and reflux for 1 hour. After the reaction, the chloroform layer was poured into ice water and washed with water to give 5-(4-chlorophenylamino)-1-
(2-chloroethyl)-1H-pyrazolo[3,4-bl
Pyrazine (Compound 2m points 163-16 of the above formula (1a)
5° C.) 2.45 g (yield 90%) are obtained.

Preparation Example 8 uses varabutylaniline instead of parachloroaniline, and Preparation Example 9 is operated in the same manner as Preparation Example 7 using varaphenetidine.

Table 1 shows the shapes and melting points of the target compounds obtained in Production Examples 1 to 9, and the results of IR and NMR analysis of these compounds.

[! 2 Preparation Example 10] 7.58 g of 1-(2-hydroxyethyl)-LH-pyrazolo[3,4-bl pyrazine (formula 10 above) was dissolved in 50 mQ of chloroform, and 5.5 g of pyridine and 8.5 g of thionyl chloride were dissolved therein. After refluxing for 1 hour, the chloroform layer was poured into ice water, washed with water, and 1-(
2-chloroethyl)-1H-pyrazolo[3,4-bl pyrazine (compound of formula (1h) above, melting point 64-65°C)
6.74 g (yield 8o%) is obtained.

3.65 g of the above 1-(2-chloroethyl)-1H-pyrazolo[3°4-b]pyrazine was dissolved in 50 d of ethanol, and 1.5 g of sodium ethoxide was added to the solution. , add 62g,
After the reaction was refluxed for 1 hour, ice water was added and extracted with benzene to obtain a compound of 1-vinyl-IH-pyrazolo[:3.4-bl pyrazine (formula 1h above), melting point 101-102].
C) 3.32 g (yield 91%) are obtained.

[Production Example 11] Dissolve 7.0 g of 5-hydroxy-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-bl pyrazine (formula 4 above) in 50+all of chloroform, and add 5.5 g of pyridine to the solution. After 6 reactions, 8.27 g of thionyl chloride was added and refluxed for 1 hour, and the chloroform layer was poured into ice water.
Wash with water and 5-hydroxy-1-(2-chloroethyl)
-1H-pyrazoloC3,4-bl pyrazine (formula (1)
Compound c), melting point 205-206'C) 6.56 g (
Yield: 85%).

The above 5-hydroxy-1-(2-chloroethyl)-1H
-Pyrazolo[3,4-b 3.97g of copyrazine was dissolved in 50% of ethanol, and 1.0% of sodium ethoxide was added to the solution.
Add 62 g and reflux for 2 hours. After the reaction, add ice water and
Acidified and extracted with chloroform to give 5-hydroxy-
1-vinyl-IH-pyrazolo[3,4-b]pyrazine (
Compound of formula (1d), melting point 233-234°C)2.
88 g (89% yield) are obtained.

[Production Example 12] The above 5-hydroxy-1-(2-chloroethyl)-1H
-Pyrazolo[3,4-bl 3.97 g of pyrazine was reacted with 20 mQ of phosphorus oxychloride, and 5-chloro-1-(2
-chloroethyl)-1H-pyrazolo[3,4-b]pyrazine (compound of formula (1e) above, melting point 79-80'C)
3.56 g (90% yield) are obtained.

4.34 g of the above 5-chloro-1-(2-chloroethyl)-1H-pyrazolo[3,4-b]pyrazine was added to 100 al.
1 of the mixture was dissolved in butanol, 2.7 g of potassium t-butoxide was added thereto, and the mixture was stirred at room temperature for 3 hours.

After the reaction, thi-butanol was distilled off, ice water was added, and extraction was performed with benzene to obtain 5-chloro-1-vinyl-IH-pyrazolo[3,4-bl pyrazine (the compound of formula (1f) above,
2.89 g (80% yield) of m.p. 55-57°C are obtained.

[Production Example 13] 3.61 g of the above 5-chloro-1-vinyl-111-pyrazolo[3°4-b copyrazine] was dissolved in 50% of ethanol, 5 mQ of hydrazine hydrate was added thereto, and the mixture was refluxed for 1 hour. After the reaction, add ice water, extract with chloroform,
5-hydrazino-1-vinyl-IH-pyrazolo[3,4
-bl pyrazine (compound of the above formula (1g), melting point 190
~191°C) 2.81g (yield 8%) was obtained.

[Production Examples 14 to 24] The above 5-chloro-1-(2-chloroethyl)-1H-pyrazolo[3,4-b]pyrazine was reacted with the compounds shown in Table 3, respectively, to achieve the purpose of Production Examples 14 to 24. The compound was obtained.

For example, in Production Example 14, 20all methanol
Dissolve 4.34 g of 5-chloro-1-(2-chloroethyl)-1H-pyrazolo[3,4-b]pyrazine in the solution, add 1.62 g of sodium methoxide, and reflux for 1 hour. and extracted with benzene to obtain 5-
Methoxy-1-vinyl-IH-pyrazolo[3,4-b]
Pyrazine (melting point 77-78°C) 3.24g (yield 92%)
).

In addition, in Production Example 24, dimethyl sulfoxide 2
5-chloro-1-vinyl-IH-pyrazolo[3
, 4-b] 3.61 g of pyrazine (formula 1f above) was dissolved, and 5.0 g of sodium paramethoxy phenoxide was added thereto.
was added and reacted for 1 hour on a boiling water bath, then ice water was added and extracted with chloroform to obtain 5-paramethoxyphenoxy-1-vinyl-IH-pyrazolo[3,4-b]pyrazine (melting point 113-134 'C) 4.56g (yield 85%
).

[Production Examples 25 to 30] 5-chloro-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (formula 8 above) was reacted with the compounds shown in Table 3, and 5- Amino-1-(2
-hydroxyethyl)-1H-pyrazolo[3,4-bl
A pyrazine derivative (formula 9 above) is synthesized, and this is reacted with thionyl chloride in the presence of pyridine to synthesize a 1-(2-chloroethyl)-5-amino-IH-pyrazolo[3,4-b]pyrazine derivative (formula 9 above). 1a) and reacted with an alkali to form 5-amino-1-vinyl-IH-pyrazolo[3
, 4-b] pyrazine derivative (1b) was obtained.

For example, in Production Example 14, 5-chloro-1-(2
-hydroxyethyl)-1H-pyrazolo[3,4-b]
3.97 g of pyrazine and 10-hexylamine were reacted in a sealed tube, excess amine was removed, and 5-hexylamino-1-(2-hydroxyethyl)-1H-pyrazolo[
3,4-b] pyrazine (compound of formula (9) above, melting point 8
0-81°C) 4.48g (yield 85%) is obtained.

2.63 g of the above 5-hexylamino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine
Dissolved in chloroform 50-, add pyridine 1.7
After adding 2.5 g of thionyl chloride and refluxing for 1 hour, the chloroform layer was poured into ice water and washed with water.
-hexylamino-1-(2-chloroethyl)-1H-
Pyrazolo[3,4-b]pyrazine (melting point 80-81°C)
Obtain 2.22 g (84% yield).

The above 5-hexylamino-1-(2-chloroethyl)-
2.75 g of LH-pyrazolo[3,4-b copyrazine is dissolved in 50% of ethanol, 1.0 Og of sodium ethoxide is added thereto, and the mixture is refluxed for 1 hour. After the reaction, ice water was added and extracted with benzene to obtain 5-hexylamino-1-vinyl-IH-pyrazolo[3,4-b]pyrazine (melting point 1
01-102°C) 1.84 g (yield 77%) is obtained.

In addition, in Production Example 30, 5-chloro-1-(2-
hydroxyethyl)-II(-pyrazolo[3,4-bl
3.97 g of pyrazine and 5.50 g of paraphenetidine were reacted in a sealed tube, excess amine was removed, and 5-(4-
ethoxyphenylamino)-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b copyrazine (compound of formula (9) above, melting point 165-166°C) 5.15 g
(yield 86%).

The above 5-(4-ethoxyphenylamino)-1-(2-
2.99 g of hydroxyethyl)-1H-pyrazolo[3,4-bl pyrazine were dissolved in chloroform 50IIIQ.

To this were added 1.7 g of pyridine and 2.5 g of thionyl chloride, and the mixture was refluxed for 1 hour. After the reaction, the chloroform layer was poured into ice water and washed with dilute hydrochloric acid and then with water to give 5-(4-ethoxyphenylamino)-1-(2-chloroethyl)-1.
H-pyrazolo[3,4-bl pyrazine (melting point 136-1
37° C.) 2.67 g (yield 84%) are obtained.

The above 5-(4-ethoxyphenylamino)-1-(2-
chloroethyl)-LH-pyrazolo[3,4-b]pyrazine (3.18 g) was dissolved in 50 mQ of ethanol, and 1.1 g of sodium ethoxide was added thereto. Add OOg and reflux for 1 hour. After the reaction, ice water was added and extracted with benzene to give 5-hexylamino-1-vinyl-IH-pyrazo a[:3.4-
b] Pyrazine (melting point 101-102'C) 1.84 g (
yield of 77%).

Table 2 shows the shape and melting point of the target compounds obtained in Production Examples 10 to 30, and the results of analysis by IR and NMR of the compound of M et al.

Next, according to experimental examples, anti-M! Tumor action and LD. shows.

[Experiment example]

A sample (compounds of Preparation Examples 1 to 30) was added to a culture medium containing 1.0xlO'/all L 5178Y cancer cells at a concentration of 100 g/-, and the mixture was incubated in air containing 5% carbon dioxide for 48 hours. After culturing, L 5178Y
The number of cells was measured using an automatic blood cell counter, and compared with Control 1 to Roll, the cell proliferation inhibition rate was determined using the following formula. The results are shown in Table 3. Also, LD in mice. I will also write about.

Cancer cell growth inhibition rate (%) = (A-B)/AX 100A
: Number of L5178YII cells in the control B: Number of L5178Y cells in the test group Note that the culture solution is 10%
RP M containing fetal bovine serum and 50 scraps/+Jl of kanamycin 1. -1640 was used.

Table 3 below shows the anti-B1. An example of a formulation of an anticancer drug is shown below.

[Example 1] Compound of Capsule Production Example 5 200g corn starch 150g talc
80g magnesium stearate 3
After thoroughly mixing 0 g or more and adjusting the particle size by passing through a 60-mesh wire mesh, the mixture is filled into 1000 gelatin capsules. It is administered orally in 1 to 3 capsules per day.

[Example 2] Compound of Capsule Production Example 8 180g silicic anhydride
Dissolve 60g magnesium stearate 5g in acetone,! l! After adding and dispersing water silicic acid, the acetone is distilled off and the mixture is granulated.

The particles are passed through a 60-mesh wire gauze to adjust the particle size, and then magnesium stearate is added and mixed to make it smooth, and the mixture is filled into 100.0 gelatin capsules.

[Example 3] Compound of Tablet Production Example 10 200g Milk M50g Corn starch 30g Magnesium stearate 5g The above components were mixed and 60 g
After passing through a mesh wire mesh and reducing the particle size to 5111,
1000 tablets are manufactured using a tablet press. This is 1
Administer 1 to 3 tablets orally per day.

[Example 4] Suppository cacao butter 1200 g Compound of Production Example 17 140 g Cocoa butter was heated to 50°C and dissolved, the compound of Production Example 14 was added thereto to make it homogeneous, and then poured into a container and cooled and solidified. to produce 1000 pieces of waste agent.

[Example 5] Compound of Injection Production Example 16 400g Polyoxyethylene hydrogenated castor oil 500g Black distilled water for injection
Total volume: 10 ml An injection is prepared according to the above-mentioned recipe in a conventional manner, and each ampoule is filled into 2 ampoules.

[Example 6] Compound of Injection Production Example 6 50 mg N'-nitoxymethyl-5-fluorouracil 200 mg Trisaminomethane 800 mg Distilled water for injection
Adequate amount Prepare an injection according to the above prescription using a conventional method.

Fill each ampoule with SmQ.

Claims (1)

[Claims] 1. The following formula (1) ▲There are mathematical formulas, chemical formulas, tables, etc.▼...(1) [However, R_1 is a hydrogen atom, a hydroxyl group, a halogen atom, a hydrazino group, -O-R' group or -NH-R' group (where R' is an alkyl group, a benzyl group, a phenyl group, or a phenyl group in which one or more hydrogen atoms of the phenyl group is substituted with a halogen atom, an alkyl group, or an alkoxy group) , R_2 is a vinyl group or -CH_2CH_2X group (
Here, X is a halogen atom). ] A 1,5-substituted-1H-pyrazolo[3,4-b]pyrazine derivative. 2. Formula (1) below ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼... (1) [However, R_1 is a hydrogen atom, hydroxyl group, halogen atom, hydrazino group, -O-R' group, or -NH-R ' group (here, R' is an alkyl group, a benzyl group, a phenyl group, or a phenyl group in which one or more hydrogen atoms of the phenyl group is substituted with a halogen atom, an alkyl group, or an alkoxy group), R_2 is a vinyl group or -CH_2CH_2X group (
Here, X is a halogen atom). 1. An antitumor agent comprising a 1,5-substituted-1H-pyrazolo[3,4-b]pyrazine derivative represented by the following as an active ingredient.