JPH02172988A - 1h-pyazolo(3,4-b)pyrazine derivative and antineoplastic agent containing the same compound - Google Patents

Abstract

NEW MATERIAL:5-(N-substituted amino)methyl-or 5-(N,N-disubstituted amino) methyl-1H-pyrazolo[3,4-b]pyrazine derivatives of formula I (R1 is H, methyl, etc.; R2 is 3-8C alkyl, phenyl, etc.). EXAMPLE:5-Hexylaminomethyl-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine. USE:A hypotoxic antineoplastio agent having an excellent antineoplastic effect. PREPARATION:With 1-(2-hydroxyethyl)-4,5-diaminopyrazole.HCl of formula II diisonitroacetone of formula III is reacted to prepare a compound of formula IV and the resultant compound is then reacted with NaNO2 to obtain a compound of formula V. The obtained compound is reacted with an amine derivative of formula R2NH2 to prepare a compound of formula VI and the resultant compound is subsequently reduced using NaBH4.

Description

Detailed Description of the Invention The present invention is based on the following formula (1) [wherein R1 represents a hydrogen atom, a methyl group, or an ethyl group, R2 represents an alkyl group having 3 to 8 carbon atoms, a phenyl group, or a halogen atom, Alkyl group having 1 to 6 carbon atoms, 1 to 6 carbon atoms
4 represents a phenyl group having an alkoxy group, a carboxyl group, or a carbethoxyl group. ] 5-(N-substituted amino)methyl- or 5-(
N,N-disubstituted amino)methyl-1H-pyrazolo(3,
4-b) Pyrazine derivative.

[However, R represents a hydrogen atom, a methyl group, or an ethyl group. R2 is an alkyl group having 3 to 8 carbon atoms, a phenyl group, or a halogen atom, an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 6 carbon atoms;
4 represents a phenyl group having an alkoxy group, a carboxyl group, or a carbethoxyl group. ] A novel 5-(N-substituted amino)methyl- or 6-(N,N-disubstituted amino)methyl-1H-pyrazoloH,4-b) pyrazine derivative and the new compound as an active ingredient. Regarding antitumor agents.

Chemotherapy agents for cancer that can be used in the future include alkylating agents, hormones, plant alkaloids, antibiotics, and antimetabolites, but all of them are highly toxic except for the antimetabolites. , L.D. is a number■
/kg to several tens of mg/-. Also, alkylating agents,
Plant alkaloids have also been shown to induce secondary cancer when administered. Most of the anti-metabolite drugs are Zorro products made from 5FU derivatives, and the problem of resistance is a major concern. Therefore, there is a strong desire to develop new anticancer drugs, and in particular,
Expectations are high for the development of new anticancer drugs among the antimetabolites, which have relatively mild effects and low toxicity.

In view of the demand for such an anticancer agent, the present inventors have previously proposed an antitumor agent containing a specific 1H-pyrazolo (3,4-bl pyrazine derivative) as an active ingredient (Japanese Unexamined Patent Application Publication No. 1983-1993).
-135810.135887゜180485.180
486,208275゜208293.59-6259
2.60-56981.181016, 181087.
No. 62-273979.63-17882), but there is a desire for a substance that has even better antitumor effects, has low toxicity, and can be effectively used as an antitumor agent.

In response to the above-mentioned request, the inventors of the present invention conducted extensive research on substances with relatively low toxicity and excellent antitumor activity, and found that 1H-pyrazolo[3,4,-b]pyrazine- 5-
5-(N-substituted imino)methyl-1H-pyrazolo(
3,4-b) By reducing the pyrazine derivative, various new compounds 5-(N=substituted amino)methyl-I H-pyrazolo(3,4-b) pyrazine derivatives described below can be obtained. By alkylating the new 5
-(N,N-disubstituted amino)methyl-1H-pyrazolo(
3,4-b) It was discovered that pyrazine derivatives can be obtained, and all of these new compounds have excellent antitumor effects and relatively low toxicity, and can be effectively used as antitumor agents. This knowledge led to the present invention.

Therefore, the present invention is based on the following formula (1). [However, R represents a hydrogen atom, a methyl group, or an ethyl group; aR2 represents an alkyl group having 3 to 8 carbon atoms, a phenyl group, or a halogen atom, an alkyl group having 1 to 6 carbon atoms, or
4 represents a phenyl group having an alkoxy group, a carboxyl group, or a carbethoxyl group. ] 5-(N-substituted amino)methyl- or 5-(
N,N-disubstituted amino)methyl-1H-pyrazolo(3,
4-b) An antitumor agent containing a pyrazine derivative and this new compound as an active ingredient is provided.

The present invention will be explained in more detail below.

Novel compound 5-(N-substituted amino)methyl- or 5-(N,N-disubstituted amino)methyl-1H according to the present invention
It is a compound having a chemical structural formula derived from -pyrazolo(3,4-b) pyrazine.

Here, R2 represents a hydrogen atom, a methyl group or an ethyl group. In addition, R2 is an alkyl group (C, H..., :n=3~
8), phenyl group or halogen atom (CI, Br),
Alkyl m (Cn R2n+, : n = 1 to 6), alkoxy group (OCnH2n+x : n = 1 to 4),
Indicates a phenyl group having a carboxyl group or a carbethoxyl group.

The N-1-substituted aminomethyl compound shown in the above-mentioned -fulfillment (1) is, for example, as shown in the following reaction formula A, 1-(2-hydroxyethyl)-4,5-diaminopyrazole.HCI (
2) is reacted with diisonitrosoacetone (3) to form 1-
(2-hydroxyethyl)-1H-pyrazolo[3,4-
b] Synthesize pyrazine-5-carboxaldoxime (4) and react with sodium nitrite to synthesize 1-(2
-hydroxyethyl)-1H-pyrazolo(3,4-b)
Synthesize pyrazine-5-carboxaldehyde (5),
Furthermore, 5-(N-fit-substituted imino)methyl-I H-pyrazolo(3,4-b) pyrazine derivative (6) is synthesized by reacting an amine derivative, and the compound is synthesized by reducing it with sodium borohydride. be able to.

Furthermore, the N,N-disubstituted aminomethyl compound can be synthesized by reacting the N-substituted methyl compound with alkyl iodide or alkyl bromide. In addition, N-substitution-
The N-methylaminomethyl compound can be synthesized by reacting formaldehyde in the presence of sodium cyanoborohydride.

CH, CH, 0H The compound of the formula (1) has excellent antitumor effects and suppresses the proliferation of cancerous cells, so it is effectively used as an antitumor agent.

In addition, when using the compound of formula (1) as an antitumor agent,
In formula (1), R2 is an alkyl group having a straight chain CnHzn
+z (however, n = 3-8), in the case of a phenyl group, the halogen atom of the substituent is 01 or Br, and the alkyl group is Cn H! n+□ (however, n=1 to 6
) and those in which the alkoxy group is OCn Hz n + (where n=1 to 4) are preferred.

The antitumor agent according to the present invention contains a compound of formula (1) as an active ingredient. It is administered by oral administration, rectal administration using suppositories, etc. The dosage varies depending on the administration route and the number of administrations, and can be varied widely depending on the severity of the symptoms, etc., but in general, the therapeutically effective dosage is 1 to 50% of the active ingredient of the present invention per 1 kg of body weight per day. ■It is.

The antitumor agent of the present invention can be prepared for administration by combining an effective amount of the compound represented by 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, a non-toxic carrier such as alcohol,
Esters, polyethylene glycol derivatives, sorbitan fatty acid esters, sulfated fatty alcohols, sorbitol, tragacanth gum, binders such as polyvinylpyrrolidone, sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate , excipients such as magnesium aluminate metasilicate, synthetic aluminum metasilicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose, magnesium stearate, talc, lubricating oil such as hydrogenated oil, 1 common salt, saccharin, Flavoring agents such as orange oil, licorice extract, citric acid, glucose, menthol, eucalyptus oil, and malic acid; suspending agents such as coconut oil, olive oil, sesame oil, peanut oil, calcium lactate, safflower oil, and soybean phospholipids; , lubricating oils, cellulose such as cellulose acetate phthalate (CAP), carbohydrate derivatives such as sugars, acrylic acid copolymers such as methyl acrylate-acrylic acid copolymers, methyl methacrylate-methacrylic acid copolymers, etc. , polyvinyl derivatives such as dibasic monoesters, other film-forming agents, coating agents, and other components by a conventional method, and then used. In addition, when preparing preparations for mucosal application, particularly suppositories, cocoa butter, lauric fat, polyethylene glycol, glycerogelatin, sodium stearate, or a mixture thereof is used as a base. Furthermore, injections are also prepared by conventional methods, but when adopting a method of suspending or emulsifying in distilled water for injection, as a suspending agent,
Sorbitan syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, aluminum stearate gel, etc. can be used, and as emulsifiers, sorbitan monooleate, polyoxyethylene hydrogenated castor oil, lecithin, etc. can be used.

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

Next, a production example of one of the compounds of the present invention will be shown.

[Production Example 1] 35.7 g of 1-(2-hydroxyethyl)-4,5-diaminopyrazole.HCI is dissolved in 150 d of methanol, and 23.2 g of diisonitrosoacetone dissolved in 50 mM of methanol is added thereto. After standing at room temperature for 24 hours, the precipitated crystals were collected by filtration to yield 24.8 g (60%) of 1-(2-hydroxyethyl)-LH-pyrazolo(3,4-b)pyrazine-5-carboxaldoxime.
is obtained.

Add 20.7g of this compound to 200aQ of concentrated hydrochloric acid or 50%
~ Dissolve in 100 mQ of 60% sulfuric acid and cool with water.

Add Sodium Nitrite Log. After addition, ice water 200ml
When diluted with d and extracted with 1000mM chloroform, 1
-(2-hydroxyethyl)-1H-pyrazolo(3,4
-b) 16.4 g of pyrazine-5-carboxaldehyde
(92%) is obtained. 1.92 g of this compound is dissolved in 50 mQ of methanol, 1.52 g of hexylamine is added, and the mixture is refluxed on a water bath for 2 hours. After the reaction, methanol 5
0d and 0.40 g of sodium borohydride, and reflux for 1 hour. Thereafter, methanol is distilled off, dissolved in 50 d of chloroform, and washed with water. Chloroform was distilled off, and the residue was recrystallized from ethanol to give 5-hexylaminomethyl-1-(2-hydroxyethyl)-18-
2.80 g (65%) of pyrazolo[3,4-b]pyrazine (colorless needle crystals, melting point 52-53 DEG C.) are obtained. Instrumental analysis data are shown in Table 2.

[Production Examples 2 to 13] In the same manner as in Production Example 1, 1-(2-hydroxyethyl)-
1H-pyrazolo(3,4-b)pyrazine-5-carboxaldehyde is synthesized and reacted with various amines shown in Table 1. Then, when reduced with sodium borohydride, 5-(N-substituted amino)methyl-1 shown in Table 1 is obtained.
-(2-hydroxyethyl)-1H-pyrazolo[3,4
-b] A pyrazine derivative is obtained. Melting point, properties, and instrumental analysis data are shown in Table 2.

[Production Example 14] 5-(4-chlorophenylamino) synthesized in Production Example 9
3.04 g of methyl-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-bl pyrazine was dissolved in 125 g of acetonitrile, and 16 g of 35% formaldehyde was dissolved therein.
．． 5d, 3.8g of sodium cyanoborohydride, and 2.5d of acetic acid were added, and the mixture was allowed to react at room temperature for 2 hours. After acetonitrile is distilled off under reduced pressure, chloroform 501d is added, and the chloroform layer is washed with I NN a OH. Dry with anhydrous sodium sulfate and chloroform is distilled off. When the residue is recrystallized from benzene + hexane (1:1),
5-(N-(4-chlorophenyl)-N-methylaminocomethyl-1-(2-hydroxyethyl)-L H-pyrazolo(3,4-b)pyrazine (yellow needles, melting point 8
8-89°C) 2°90g (91%) is obtained. Equipment data is shown in Table 2.

[Production Example 15] 5-(4-butylphenylamino) synthesized in Production Example 7
3.25 g of methyl-1-(2-hydroxyethyl)-1H-pyrazolo(3,4-b)pyrazine was added to 10 g of methanol.
0d, and a reaction mixture of 50all methanol and 0.5g of metallic sodium is added thereto. Further, 3.0 g of methyl iodide was added and the mixture was refluxed for 12 hours. After the reaction, methanol is distilled off. Dissolve the residue in chloroform and wash with water.

Dry with anhydrous sodium sulfate. After that, chloroform was distilled off and the precipitated crystals were recrystallized from benzene.
5-(N-(4-butylphenyl)-N-methylaminocomethyl-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-b]pyrazine (yellow needles, melting point 68
~71° C.) 2.34 g (69%) are obtained. Equipment data is shown in Table 2.

(Production Example 16) 5-(4-butylphenylamino) synthesized in Production Example 7
3.25 g of methyl-1-(2-hydroxyethyl)-1H-pyrazolo(3,4-b)pyrazine was dissolved in 10 ml of ethanol.
A mixture of 50 mQ of methanol and 0.5 g of sodium metal was added to the solution. Furthermore, 3.0 g of methyl bromide was added and the mixture was refluxed for 12 hours. After the reaction,
Distill ethanol away. Dissolve the residue in chloroform,
Wash with water and dry with anhydrous sodium sulfate. Thereafter, chloroform was distilled off, and the precipitated crystals were recrystallized from benzene + hexane (1:1), resulting in 5-CN-(4-butylphenyl)-N-ethylaminocomethyl-1-(2-hydroxyethyl )-1H-pyrazolo(3,4-b)pyrazine (yellow needle-shaped crystals, melting point 50-52°C) 2.40 g
(68%) is obtained. Equipment data is shown in Table 2.

Table 2 (note) Properties C: colorless, Y: yellow, Ly: pale yellow, N: acicular.

Pr niblistic, Sc: scaly solvent H: hexane, B: benzene 9M: methanol.

C: Chloroform MR S: Singlet, B S: Broad
-5inglet.

D: Doublet, T: Triplet, M
: Multiplet.

Q: Quartet Next, based on experimental examples, the antitumor effect and L
D. shows.

[Experimental Example 1] A sample (compounds of Preparation Examples 1 to 16 above) was added to a culture medium containing 1.0 x 10'/- L1210 mouse leukemia cells so that the final concentration was 1100 p/mQ.
After culturing at 37° C. for 48 hours in air containing % carbon dioxide gas, the number of viable cells was measured, and compared with a control measured in the same manner, the cell growth 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)/AX100
A = Number of L12101 [1 cell in control] B: Number of L1210 cells in test group The culture medium was 10% fetal bovine serum and 50 A.
RPM11640 containing kanamycin of /aQ was used.

Table [Experimental Example 2] The antitumor effect (in viv
o) was investigated.

J c 1- I CR 6 weeks old, female mice (- group 6 mice)
I x 10' Sarcoma 180 cancer cells were implanted intraperitoneally, and 24 hours later, the compound dissolved in Lipiodol (
100 mg/kg) was administered intraperitoneally for 5 consecutive days.

Thereafter, the survival days of the mice were determined wt, and the median survival days were taken as the survival days of that group, and the survival days of the control group (administered intraperitoneally with Lipiodol alone for 5 consecutive days) determined in the same way was taken as 100. The life-prolonging effect of the compound is 11mL.
The results are shown in Table 4.

Table 4 Examples of formulations of the antitumor agent of the present invention are shown below in Examples.

[Example 1] Compound of Capsule Production Example 1 200g corn starch 150g talc
At least 30 g of 80 g magnesium stearate is thoroughly mixed, passed through a 60-mesh wire gauze to adjust the particle size, and then filled into 1000 gelatin capsules. It is administered orally in 1 to 3 capsules per day.

[Example 2] Compound of Capsule Production Example 1 180g silicic anhydride
150g magnesium stearate
5g of the compound of Production Example 1 is dissolved in acetone, silicic anhydride is added and dispersed, and then the acetone is distilled off and granulated. The particles are passed through a 60-mesh wire mesh to adjust the particle size, and then magnesium stearate is added and mixed to make it smooth, and the mixture is filled into 1000 gelatin capsules.

[Example 3] Tablet Compound of Tablet Production Example 15 200g Lactose
50g corn starch aog magnesium stearate
5 g of each of the above ingredients are mixed, passed through a 60-mesh wire mesh to adjust the particle size, and then manufactured into 1000 tablets using a tablet press. It is administered orally in 1 to 3 tablets per day.

[Example 4] Suppository cacao butter 1200 g Compound of Production Example 16 140 g Cocoa butter was heated to 50°C and dissolved, the compound of Production Example 16 was added thereto and made uniform, then poured into a container and cooled. Solidify to make 1000 suppositories.

(Example 5) Injection Compound of Production Example 16 Polyoxyethylene hydrogenated castor oil Distilled water for injection Total amount 0ON 500■ 0mQ Fill 1 ampoule 2 times each.

[Example 6] Compound 5001g of Injection Production Example 16 Trisaminomethane 800g Applicant Lion Co., Ltd. Agent Patent Attorney Takashi Kojima Academic Continuation Amendment (Spontaneous) November 9, 1989 1, Display of the case 1985 Patent Application No. 327354 2, Name of the invention 1H-pyrazolo (3,4-bl pyrazine derivative and antitumor agent containing the compound 3) Person making the amendment Address related to the case Name

Claims (1)

[Claims] 1. The following formula (1) ▲There are mathematical formulas, chemical formulas, tables, etc.▼...(1) [However, R_1 represents a hydrogen atom, a methyl group, or an ethyl group. R_2 is an alkyl group having 3 to 8 carbon atoms, a phenyl group,
Alternatively, it represents a phenyl group having a halogen atom, an alkyl group having 1 to 6 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, a carboxyl group, or a carbethoxyl group. ] 5-(N-substituted amino)methyl- or 5-(
N,N-disubstituted amino)methyl-1H-pyrazolo[3,
4-b] Pyrazine derivative. 2. Antitumor containing the 5-(N-substituted amino)methyl- or 5-(N,N-disubstituted amino)methyl-1H-pyrazolo[3,4-b]pyrazine derivative according to claim 1 as an active ingredient. agent.