JPH02240084A - 1,5-disubstituted-1H-pyrazolo[3,4-b]pyrazine derivatives and antitumor agents containing the compounds - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the following formula (1) [wherein R represents a hydrogen atom, a methyl group, an ethyl group, a 2-hydroxyethyl group, a benzyl group, a vinyl group, or a phenyl group. Moreover, R2 is a hydrogen atom, a cyano group, or C n
Straight chain alkyl group (n
-1 to 17). ] 1,5-disubstituted-IH-pyrazolo[3,4-
b] Virazine derivative.

(However, R1 represents a hydrogen atom, a methyl group, an ethyl group, a 2-hydroxyethyl group, a benzyl group, a vinyl group, or a phenyl group. Also, R2 represents a hydrogen atom, a cyano group, or a CnH
Straight chain alkyl group represented by 2n+iCO (n-
1 to 17). ] Novel 1,5-disubstituted-IH-pyrazolo (
3.4-b) Relating to an antitumor agent containing a pyrazine derivative and the new compound as an active ingredient. The growth is
Chemotherapy agents for cancer include alkylating agents, hormones, plant alkaloids, antibiotics, and antimetabolites, but all of these conventional chemotherapeutic agents are toxic except for the antimetabolites. Strongly, LDo is a number IlIg/
'kx to several tens of kg/kg. Also, alkylating agents,
Plant alkaloids have also been shown to induce secondary cancers when administered. Most of the anti-metabolite products are 5FU derivatives, and the problem of resistance has become an important issue. Therefore, under these circumstances, there is a strong desire for the development of new anticancer drugs, and in particular, there are high hopes for the development of new anticancer drugs among the antimetabolites, which have relatively mild action but low toxicity. There is.

In view of the demand for such an anticancer agent, the present inventor has previously proposed an antitumor agent containing a specific IH-vilazolo (3,4-bl virazine derivative) as an active ingredient (JP-A-58
-135810.135887, 180485.180
4B6.208275, 208293, 59-6259
2, 60-56981, 181016.181087,
62-273979, 63-17882), but there is a need for more effective antitumor agents.

In order to meet the above-mentioned needs, the inventors of the present invention conducted extensive research on substances with relatively low toxicity and excellent antitumor activity, and found that 5-chloro1-monosubstituted-IH -Birazoro (
3,4-b) Reacting pyrazine derivative with sodium sulfide to obtain S-mercabuto 1-monosubstituted-IH-virazolo (3,4-b)
4-1): 5-acylthio-1-monosubstituted-IH-virazolo[3
, 4-b) Pyrazine derivatives can also be converted to 5-thiocyano-1 monosubstituted-IH by reacting with promocyan.
- It was discovered that pyrazolo(3.4-b)virazine derivatives can be obtained, and that all of these new compounds have excellent antitumor effects, have relatively low toxicity, and are effective as antitumor agents. The inventors have discovered that the present invention can be used.

Therefore, the present invention provides the following formula (1) R1 [wherein R1 represents a hydrogen atom, a methyl group, an ethyl group, a 2- and droxyethyl group, a benzyl group, a vinyl group, or a phenyl group. Moreover, R2 is a hydrogen atom, a cyano group, or C n
Straight chain alkyl group (n
=1 to 17). ] 1,5-disubstituted-IH-virazolo[3,4-
b] To provide an antitumor agent containing a virazine derivative and this new compound as an active ingredient. The present invention will be explained in more detail below.

The novel compound 1,5-disubstituted-IH-virazolo(3.4-b) virazine derivative according to the present invention has the above-mentioned formula (1),
That is, it is a compound having the chemical structural formula shown below. here,
Ri represents a hydrogen atom, a methyl group, an ethyl group, a 2-hydroxyethyl group, a benzyl group, a vinyl group, or a phenyl group. Further, R2 is a hydrogen atom, a cyano group, or a C. ,Hz
Straight chain alkyl group represented by n + z C O (n
6 representing an acyl group having =1 to 17) above general formula (1)
For example, the 1,5-disubstituted product represented by
As shown in Figure 3, 5-grollo-1 monosubstituted-IH-virazolo[3.4-b:l pyrazine derivative (2) was reacted with sodium sulfide to produce 5-mercapto-1-monosubstituted-IH-virazolo[3. 3.4-b) Synthesize pyrazine derivative (1a) and react with acid anhydride or acid chloride to synthesize 5-acylthio-1-monosubstituted-IH-pyrazolo (3.4-b). A pyrazine derivative (1b) was obtained, and the sodium salt (la-N a ) of Doki derivative (1a) was reacted with promosyan to give 5-thioneano-1 monosubstituted-
IH-pyrazolo (3,4-b:l virazine derivative (1c
) can be synthesized by obtaining.

Reaction formula A RL (la-Na) (IC) The compound of formula (1) has an excellent antitumor effect and inhibits the proliferation of cancerous cells, so it is effectively used as an antitumor agent.

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 alone or in combination with other pharmaceutical ingredients if necessary. ．． It is administered by methods such as subcutaneous injection, oral administration, and rectal administration using suppositories. The dosage varies depending on the route of administration 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 kg of body weight per day.
The amount of the active ingredient of the present invention is 1 to 50 per day.

The antitumor agent according to the present invention can be prepared by combining an effective amount of the compound represented by the general formula (1) with an appropriate amount of non-toxic carrier and preparing it for administration using any conventional formulation method.

That is, when preparing for oral administration, it can be prepared into soft capsules, hard capsules, tablets, granules, fine granules, powders, active ingredient sustained release agents, solutions, suspensions, etc. When administering parenterally, For injections, drips, suppositories, etc. It can be made by iliI.

In this case, when formulating, non-toxic carriers such as alcohols, esters, polyethylene glycol derivatives, sorbitan fatty acid esters, sulfated fatty alcohols, binders such as sorbitol, gum tragacanth, polyvinylpyrrolidone, sucrose, lactose, Excipients such as starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium aluminate metasilicate, synthetic aluminum metasilicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose, stearin Magnesium acid, talc, lubricating oil such as hydrogenated oil, salt, saccharin, orange oil, licorice extract, citric acid, glucose, menthol, eucalyptus oil, flavoring agents such as malic acid, flavoring agent, coconut oil, olive oil. Sesame oil, peanut oil, calcium lactate,
Suspending agents such as safflower oil and soybean phospholipids, lubricating oils, cellulose such as cellulose acetate phthalate (CAP), carbohydrate derivatives such as sugars, methyl acrylate, acrylic acid copolymers, methyl methacrylate, methacrylic acid It is prepared by a conventional method using components such as acrylic acid copolymers such as copolymers, polyvinyl derivatives such as dibasic acid monoesters, other film forming agents, and coating agents, and is used. In addition, when preparing preparations for mucosal application, especially suppositories, cacao butter, lauric fat, polyethylene glycol, glycerogelatin, sodium stearate, or a mixture thereof can be used as the base material. Furthermore, injections can also be prepared by conventional methods, but when a method of suspending or emulsifying in distilled water for injection is adopted, sorbitol syrup, methylcellulose,
Gelatin, hydroxyethylcellulose, aluminum stearate gel, etc. can be used, and as emulsifiers, sorbitan monooleate, polyoxyethylene hydrogenated castor oil, lecithin, etc. can be used.

The novel compound represented by the above formula (1) of the present invention has an antitumor effect and is therefore useful as an antitumor agent, and an antitumor agent containing this new compound as an active ingredient has excellent antitumor activity. In addition, it has relatively low toxicity and is effective against solid cancers.

Next, production examples of the compounds of the present invention will be shown.

[Production Example 1] 5-chloro-1-phenyl-IH-bilazolo (3.4
- b) Pyrazine 4.60 g. Sodium sulfide 6
．． 30 g is dissolved in dimethyl sulfoxide 100- and reacted for 1 hour on a boiling water bath. After the reaction, the reaction mixture is poured into ice water and acidified with hydrochloric acid, and the precipitated crystals are collected by filtration. After drying the filtered crystals, recrystallization from chloroform yields 5-mercapto-1-phenyl-1H-vilazolo (
3.4-b) Pyrazine (yellow-brown needle crystals, melting point 215-
219°C) 4.0g (88%) is obtained. The compound of Production Example 2 shown in Table 1 is also obtained in the same manner.

[Production Example 3] 5-chloro-1-vinyl-IH-virazolo (3.4
- b) 5.40 g of pyrazine, 9.50 g of sodium sulfide
Dissolve g in 150 d of dimethyl sulfoxide and react on a boiling water bath for 1 hour. After the reaction, the reaction mixture is poured into ice water, acidified with hydrochloric acid, and extracted with ethyl acetate. The quasi-ethyl acetate layer was dried over anhydrous sodium sulfate, the ethyl acetate was distilled off, and the precipitated crystals were recrystallized from chloroform.
5-Mercapto-1-vinyl-IH-pyrazolo (3.4
1b] Pyrazine (yellow-red needle crystals, melting point 186-18
7° C.) 4.9 g (92%) are obtained.

The compound of Production Example 4 shown in Table 1 is also obtained in the same manner.

[Production Example 5] 5-chloro IH-pyrazolo (3.4-b) pyrazine 3
．． 10 g of sodium sulfide and 6.30 g of sodium sulfide were dissolved in 100 g of dimethyl sulfoxide and reacted for 5 hours on a boiling water bath. After the reaction, the reaction mixture was poured into ice water,
Acidify with hydrochloric acid and filter the cut out crystals. After drying the filtered crystals, the crystals were recrystallized from ethanol to yield 2.9 g (88%) of 5-mercapto-IH-pyrazolo(3.4-b)virazine (reddish brown flaky crystals, melting point 250-251°C).
) is obtained.

[Production Example 6] 5-mercapto-1-vinyl-IH-virazolo (3.4
-b) Add 10 mQ of acetic anhydride and 0.1 g of sodium carbonate to 1.78 g of pyrazine and reflux for 1 hour. After the reaction, excess acetic anhydride was distilled off, and the residue was dissolved in benzene.
Wash with S% sodium bicarbonate solution. The benzene layer is dried over anhydrous sodium sulfate, and the benzene is distilled off. The obtained crystals were recrystallized from hexane to yield 2.1 g of 5-acetylthio-1-vinyl-IH-virazolo(3.4-b)virazine (colorless needle-like crystals, melting point 102-103°C).
(95%) is obtained.

The compound of Production Example 7 shown in Table 1 is also obtained in the same manner.

[Production Example 8] 5-mercapto-1-vinyl-IH-vilazolo (3.4
-b) Pyrazine 1.78 g, lauroyl chloride 2.3
0g. 0.2 g of triethylamine and 20 g of chloroform
Dissolve in mQ and reflux for 1 hour. After the reaction, chloroform and excess triethylamine are distilled off, and the residue is dissolved in benzene and washed with 5% sodium hydrogen carbonate solution. The benzene layer is dried over anhydrous sodium sulfate, and the benzene is distilled off. Recrystallization of the residue from hexane yields 5-lauroylthio-1-vinyl-I H-pyrazolo[3.4-b]
Pyrazine (colorless needle-like crystals, melting point 55-56°C) 2.6g
(72%) is obtained.

The compound of Production Example 9 shown in Table 1 is also obtained in the same manner, [Reverse Production Example 10] S-mercapto-1-phenyl-IH-pyrazolo (3.
4-bl pyrazine 2.2 8 E is dissolved in 120 mQ of NaOH methanol solution (containing 0.80 g of NaOH), and an ethanol solution 10 of 1.10 g of cyanogen bromide is added dropwise while keeping the temperature at 0°C. Furthermore, the reaction was carried out while maintaining the temperature at 0° C. for 30 minutes. After the reaction, ice water was added, and the precipitated crystals were collected by filtration, washed with water, dried, and recrystallized from hexane to give S-thiocyano-1-phenyl-IH-vilazolo (
3.41b] Pyrazine (pale yellow scaly crystals, melting point 128
-129℃) 2.30g (91%) is obtained, the first
The compound of Production Example 11 shown in the table is also obtained in the same manner. Table 2 shows the characteristic values of the compounds in Table 1 obtained by the above method.

Next, the antitumor effect of the present invention and LD. shows.

[Experiment Example 1] 1. Add the sample (compounds of Preparation Examples 1 to 16 above) to a culture solution containing 105 OX/IILQ L1210 mouse leukemia cells at a final concentration of 100x/1,
After culturing at 37° C. for 48 hours in air containing 5% carbon dioxide gas, the number of viable cells was measured, and compared with Control 1 and Roll, which were measured in the same manner, 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)/AxlOOA
: Number of L1210 cells in control B : Number of L1210 cells in test group The culture medium was 10% fetal bovine serum and 50 t
[Experimental Example 2] Using RPMI 1640 containing kanamycin, the antitumor effect (in vivo) of the compound of the production example against Sarcoma 180 was investigated by the following method.

JCL-ICR 6 week old female mice (6 mice per group) were injected with Sarcoma 180 cancer cells 1. Transplant OX10 into the abdominal cavity,
After 24 hours, the compound dissolved in Libiodol (100
/kg) was administered intraperitoneally for 5 consecutive days. Thereafter, the number of survival days of the mice was observed, and the median number of survival days was taken as the survival number of that group, and the survival number of the control group (intraperitoneally administered with only Lipiodol for 5 consecutive days) determined in the same manner was set as 100.
The compound was found to have a life-prolonging effect. The results are shown in Table 4.

No. table 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
80g magnesium stearate
At least 30 g of the mixture is thoroughly mixed, passed through a 60-mesh wire gauze to adjust the particle size, and then filled into 100 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 Dissolve the compound of Production Example 1 in acetone, add silicic anhydride and disperse, then distill off the acetone and granulate. . 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 the particles smooth and filled into 1000 gelatin capsules.

[Example 3] Tablet Compound of Production Example 10 200g Lactose 50g Corn starch 30g Magnesium stearate 5g The above ingredients were mixed and passed through a 60-mesh wire gauze to adjust the particle size. Manufacture 1000 tablets. It is administered orally in 1 to 3 tablets per day.

[Example 4] Suppository cacao butter 1200 g Compound of Production Example 8 140 g Cocoa butter was heated to 50°C and dissolved, the compound of Production Example 8 was added thereto to make it homogeneous, then poured into a container and cooled. Solidify to make 100 suppositories.

[Example 5] Injection Preparation Compound of Example 8 400 mg Distilled water for injection Total amount 10 mQ Prepare an injection according to the above recipe in a conventional manner, and fill 1 ampoule of 2 d each. [Example 6] Compound of Injection Preparation Example 8 50 ■ Tris-aminomethane 800 ■ Distilled water for injection Appropriate amount In accordance with the above recipe, an injection is prepared by a conventional method and filled into 1 ampoule (5-5 mm).

Applicant: Laion Co., Ltd. Agent: Patent Attorney Takashi Kojima

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 methyl group, an ethyl group, 2-
Indicates a hydroxyethyl group, benzyl group, vinyl group or phenyl group. In addition, R_2 is a hydrogen atom, a cyano group, or a C
An acyl group having a straight chain alkyl group (n=1 to 17) represented by _nH_2_n_+_1CO. ] 1,5-disubstituted-1H-pyrazolo[3,4-
b] Pyrazine derivative. 2. An antitumor agent containing the 1,5-disubstituted-1H-pyrazolo[3,4-b]pyrazine derivative according to claim 1 as an active ingredient.