JPS60130521A - Anticancer agent - Google Patents

Abstract

PURPOSE:An anticancer agent containing an indeno[1,2-c]pyrazole derivative as an active constituent, and having a low toxicity, particularly very weak inhibitory action on myeloma unique to the anticancer agent. CONSTITUTION:An anticancer agent containing a compound expressed by the formula (R<1> is H or methyl; R<2> is phenyl or pyridyl which may or may not have a substituent group; X is carbonyl or methylene which may or may not have a substituent group) as an active constituent. The dosage form is a tablet, capsule, granule or syrup, etc. for oral administration or injection or suppository, etc. for parenteral administration. The dose thereof is 20mg-1g/day/man for oral administration and 2-300mg/day/man for parenteral administration. The compound expressed by the formula (R<1> is H; X is carbonyl) is synthesized by reacting the corresponding 2-acyl-1,3-indanedione with hydrazine.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an anticancer agent containing an indeno(1,2-c) pyrazole derivative as an active ingredient.

In recent years, many anticancer drugs have been developed and are being applied clinically. On the other hand, the mortality rate due to cancer is increasing year by year, indicating that conquering cancer is still an unresolved issue for humanity.

Although some cancers are effective against conventional anticancer drugs, some cancers are difficult to respond to, including due to the development of resistance, and this is one of the reasons why cancer mortality rates cannot be lowered. Therefore, research into anticancer drugs is becoming more and more active, particularly by partially modifying the chemical structure of conventional anticancer drugs such as pyrimidine, nitrosourea, or adriamycin derivatives to develop more potent or less toxic anticancer drugs. Many attempts have been made to develop it. However, even if the anticancer drugs developed in this way have a higher therapeutic index in animal experiments, it is difficult to imagine that they will have a significant effect on cancer patients who are difficult to respond to conventional anticancer drugs. That is, for cancer patients for whom conventional anticancer drugs have difficulty demonstrating their usefulness, it is possible that a different treatment method or a new type of anticancer drug may be effective. Based on this idea, the present inventors have conducted research to find an anticancer drug that is expected to have a chemical structure completely different from that of conventional anticancer drugs, and therefore a mechanism of action different from that of conventional anticancer drugs.

As a result, indeno(1,2-
c) The present invention was completed by discovering that pyrazol-4(IH)-ones and their derivatives not only exhibit strong anticancer effects, but also extremely low toxicity, particularly extremely weak bone marrow suppressive effects specific to anticancer agents.

There have been no reports to date that indeno Ij, 2-c) pyrazole derivatives have anticancer activity. For example, Lemkθ et al.
-butyl-4-aryl-4-hydroxyindeno(1
Although the results of a preliminary study on the effect of ,2-cl pyrazole on Ehrlich ascites cancer have been reported, they have not shown a statistically significant anticancer effect CT, L,
I+emke, L, A, Cate, s, M, S
teeh-berg, and Y. M. Cho.
, r, Pharm, Sci,, 64.1575 (
+975)). In fact, the inventors' inch/[1°2-
〇〇 Research on the relationship between the chemical structure and anticancer activity of pyrazole derivatives has also shown that in the case of the R1 canicyl group of the following general formula (1), similar to the results of Lemce et al. No anticancer effect was observed. Let us further discuss the chemical structure and anticancer effect. C. Not only when n+ is an ethyl group, but also when n+ is a propyl group, a butyl group, a hydroxyethyl group, a chloroethyl group, etc. Even in the case of an alkyl group, an aryl group such as a phenyl group or a pyridyl group with or without a substituent, or an aralkyl group, a remarkable anticancer effect is not observed, and R1 is hydrogen for the expression of an anticancer effect. It turns out that being an atom or a methyl group is a chemical structural requirement.

Furthermore, when R1 is a hydrogen atom, an alkyl group with or without a substituent, or an aralkyl group, no significant anticancer effect is observed, and a phenyl group with or without a substituent, or a pyridyl group with or without a substituent. It has anticancer activity only in this group.

Furthermore, effectiveness is exhibited when X is a carbonyl group or a methylene group with or without a substituent.

That is, the present invention has a strong anticancer effect that could not be predicted from the report by Lemkθ et al.
Regarding toxicity, indeno (1,
2-=c:) This relates to an anticancer agent containing a pyrazole derivative as an active ingredient, and is related to the general formula (1) (wherein R is a hydrogen atom or a methyl group, and R is a phenyl group or a pyridyl group with or without a substituent. or a carbonyl group or a methylene group with or without a substituent).

General formula (1) used as an active ingredient of the anticancer agent of the present invention
) are shown in Table 1.

1B Among the compounds represented by the general formula (1), the general formula (in the formula,
The compound represented by R2 represents a phenyl group or a pyridyl group with or without a substituent is described by Brown and Mosher (R, A, Braun and W, A,
Mo5her, J, Wholesale, Chem, Sac, ,
80.4919 (1958)] by the reaction of the corresponding 2-acyl-1-ro-indanedione with dorazine.

Among the compounds represented by the general formula (1), the general formula (in the formula,
R2 is the same as above) is a known compound (R, A
, Braun and W., A., Mo5her,
, T, Org, Ohem, .

It can also be synthesized by converting the corresponding 6-aryl-indeno(1,2-c) pyrazole into a sodium salt or potassium salt and reacting it with methyl halide according to the method of Vol. 24, p. 648 (+959). Do 2-
It can be synthesized in one step by reaction of acyl-1,3-indanedione and methylhydrazine.

Among the compounds represented by the general formula (1), the general formula (in the formula,
R1 is a hydrogen atom or a methyl group, R2 is a phenyl group or a pyridyl group with or without a substituent, R3
represents a hydrogen atom or a substituent such as a hydroxyl group, an amide group, an alkoxy group, an alkylamino group, etc.) is synthesized, for example, as follows.

R575 (hydroxyl group compound can be obtained by reducing the corresponding indeno[1<2-c]pyrazol-4(+H)-one) with sodium borohydride.

A compound in which R3 is a hydrogen atom is synthesized by treating a compound in which R3 is a hydroxyl group with thionyl chloride to give 4-chloroindeno[1°2-0]pyrazole, which is then reduced with lithium aluminum hydride. Also, publicly known (D,
A. Habeck and W. J. Houli
Han, Ger.

It can also be obtained by the method of 0ffen, 2,517,7+6N974)). 4-chloroindeno [1,
2-c) When pyrazole is heated in lower alcohol, R
A compound in which 3 is a lower alkoxy group is obtained, and upon reaction with an alkylamine, a compound in which R3 is an alkylamino group is obtained. Compounds in which R3 has 7 ami groups or 7 alkyl ami groups are known (B, Loev, U, S).

It can also be synthesized according to the method of [Patent 3+ 004,983 (1960)].

Synthesis examples of typical compounds used as active ingredients of the anticancer agent of the present invention are shown below.

Synthesis Example 1 0.1 mol of 2-acyl-1,6-indandione was added to 25
Dissolved in 0-500 me ethanol and a catalytic amount of p-
) Add luenesulfonic acid and 0.2 mol of hydrazine hydrate and heat under reflux for 6-7 hours.

After cooling, the precipitate was recrystallized from ethanol, benzene, methyl cellosolve, or dimethylformamide to obtain compound number L 2.3. Compounds with proboscis 5.6.7.8.9 are obtained in yields of +4.8-78.7%.

Synthesis example 2 2-arylmethylidene-1,6-indanedione 05
mol, p-toluenesulfonyl hydrazide 0.55 mol, p-)luenesulfonic acid 249 and butanol 1.
The mixture of No. 91 was heated under reflux for 10 hours. The precipitate that will be generated is reconstituted from ethanol or methyl cellosolve to form compound number 4゜10, +1. ← 15 compounds of society
．． 2-67.4% yield.

Synthesis Example 6 0.2 mol of 2-acyl-1,6-indanedione was added to 0.2 mol.
5-11 in ethanol, a catalytic amount of p-toluenesulfonic acid and 0.4 mol of methylhydrazine were added, and the mixture was heated under reflux for 6-7 hours. The precipitate that will precipitate in the future is purified by the fractional crystallization method to obtain compound number 12.13i 4,15,1
6,17i 8i 9.2 + compound 21.5-7
Obtained with a yield of 8.9%.

Synthesis Example 4 Compound 20p of Compound No. 12 was dissolved in 200me of concentrated sulfuric acid, and nitric acid was added to the solution while maintaining the temperature below -5°C.
Add 87 gradually. After stirring for 1 hour, the reaction mixture was poured into water and the resulting precipitate was collected. 1-Methyl-6-(4-nitrophenyl)inch/(4,2-c
]Pyrazol-4(+H)-one (Compound No. 30) 8
．． Get 6P. Next, compound 5y of compound number 60 was added to 5
Dissolve in 200me of 0% sulfuric acid and add 6y of iron powder to 60-7
Stir at 0°C for 24 hours. After pouring the reaction mixture into water to remove insoluble matter, a 10% aqueous sodium hydroxide solution was added to the p solution to make it alkaline. The resulting precipitate is recrystallized from methyl cellosolve to obtain compound No. 20 (0.8y).

Synthesis Example 5 Corresponding indeno(1,2-c)pyrazole-4(1H
0.1 mol of )-one is dissolved in 500 m/ml of ethanol, 5-0.2 mol of sodium borohydride O,I is added, and the mixture is heated under reflux for 4-6 hours. After distilling off the solvent, add a small amount of water, extract with chloroform, dry the chloroform layer, distill it off, and recrystallize the residue from benzene, ethanol, or a mixture thereof to obtain compound number 24.25.26.
The compound is S+, +-9S.

Obtained with a yield of 4%.

Synthesis Example 6 4-Hydroxyindeno(1) obtained according to Synthesis Example 5
, 2-c:) 60me of thionyl chloride is added to 0.1 mol of pyrazole and heated under reflux for 6 hours. After distilling off excess thionyl chloride, the residue was washed with ether to obtain crude 4-
Chloroindeno(1,2-c)pyrazole 92.5-
Obtained with a yield of 94.0%.

4-chloroindeno(+,
2-c) After dissolving 50 mmol of pyrazole and decomposing the 50 lithium aluminum oxide, the solution is acidified with hydrochloric acid and extracted with chloroform. The extract is dried and the solvent is distilled off. The residue is recrystallized from ethanol to obtain compound No. 22.25 with yields of 78.7 and 41.8%, respectively.

Synthesis Example 7 Compound 5.5y of Compound No. 19 is added to 70me formic acid amide and heated under reflux for 1 hour. The precipitate that will precipitate out is washed with ethanol and then ether, 25% hydrochloric acid 46me is added, and the mixture is heated under reflux for 5 hours. Remove the insoluble matter when heated, concentrate the p solution to half its volume, and leave it in a cool place. Take the precipitated crystals and recrystallize them from water to obtain 0.85y of compound No. 27.
obtained as the hydrochloride.

Synthesis Example 8 1-Methyl-6-phenyl- obtained according to Synthesis Example 6
4-chloroindeno Ij, 2-Q) pyrazole 0, 1
mol was dissolved in anhydrous benzene filter, 0.22 mol of alkylamine was added thereto, and the mixture was heated under reflux for 15 hours.

Compound No. 2 was extracted from the reaction mixture according to a conventional method.8. 2
The free amine or hydrochloride salt of 9 is obtained in a yield of 556-60.7%.

Next, the anticancer effects of typical compounds used in the anticancer agent of the present invention will be described.

(1) Effect of intraperitoneal administration on allogeneic tumors The study drug was administered intraperitoneally 5 times every other day from the 1st day after 6 x 10 cells of Sarcoma 180 cells, which are allogeneic tumors, were subcutaneously implanted in the lumbar region of 7-8 ICE mice in one group. administered.

On the 21st day after transplantation, the tumor was excised, its weight was measured, and the tumor growth inhibition rate (%) was calculated using the following formula.

The results are shown in Table 2. As is clear from the table, these compounds markedly inhibit the growth of allogeneic tumors when administered intraperitoneally.

Table 2 (2) Effect of oral administration on homogeneous tumors in Group 1, 7 animals
3X+06 sarcoma 180 subcutaneously in the lumbar region of R mouse
The cells were transplanted, and the test drug was orally administered once a day for 10 consecutive days starting the day after transplantation. On the 21st day after transplantation, the tumor was excised and weighed, and the tumor growth inhibition rate ($) was calculated using the above formula.

The results are shown in Table 6. As the table shows, these compounds markedly inhibit tumor growth even when administered orally.

(B) Growth inhibition effect on syngeneic tumors 107 cells of breast cancer MM102, which is a syngeneic tumor of the same mice, were subcutaneously implanted into the lumbar region of 7 C3H/He mice in group 1, and the test drug was intraperitoneally administered 5 times every other day from the next day. did.

On the 5th day after transplantation, the tumor was excised, its weight was measured, and the tumor growth inhibition rate was calculated from the above formula.

The results are shown in Table 4. As is clear from the table, these compounds have a remarkable tumor growth inhibiting effect not only on allogeneic tumors but also on syngeneic tumors.

Table 4 (4) Survival prolongation effect in syngeneic tumors 107 cells of a syngeneic tumor, breast cancer MM102, were subcutaneously implanted in the lumbar region of 10 c3H/He mice in group 1, and the test drug was administered intraperitoneally 5 times every other day starting from the next day. The life and death of tumor-bearing mice were observed for 60 days after transplantation, and the effectiveness was determined based on the average survival days of each experimental group and the survival rate when the control group was set as 100.

The results are shown in Table 5. As is clear from Table 5, these compounds exhibit a significant survival effect on syngeneic tumors at doses of +0-40 mg/Ag.

Table 5 shows the toxicity of typical compounds used in the anticancer agent of the present invention.

(1) Acute toxicity in mice Test drug 1000.500 was administered to 5 male engineered CR mice per group.
and 250 my/kl? was administered intraperitoneally or orally at 2000.1O00 and 500 mg/Iv, and the progress was observed for 7 days. 1000 and 50
In the case of intraperitoneal administration at 0 my/kg, sedation and piloerection were observed immediately after administration depending on the test drug, but recovery occurred 1 hour after administration, and no deaths were observed in any of the experimental groups. In the case of oral administration, no findings that appeared to be toxic symptoms due to the test drug were observed. In other words, as shown in Table 6, the LD5Q values of these compounds in mice are >+000 my/ky when administered intraperitoneally, and >2000 my/ky when administered orally.
It is more than oomg/kg.

(2) Acute toxicity Test drug 3000.20 to 5 male and female CD rats per group
00 and + 000 my/kg were orally administered and the progress was observed for 7 days. 3000 and 2000+++g/
Other than the presence of the administered drug in the feces of the lv administration group, no findings such as toxic symptoms that were considered to be due to the administration of the test drug were observed. That is, when these compounds are orally administered to rats, the LD5Q values are 3000 mf1kg or more for both males and females, as shown in Table 7.

Table 7 (6) Myelotoxicity Test drug 400 mg/lv to 5 male CD rats in 1 group
was orally administered once a day for 5 consecutive days, and bone marrow (femur) and peripheral blood were collected on the 6th day. The bone marrow was fixed with 10% neutral buffered formalin solution, paraffin sections were prepared, hematoxylin-eosin staining was performed, and microscopic examination was performed. In peripheral blood, the number of red blood cells, hemoglobin, hematocrit, white blood cells, and platelets were measured according to conventional methods. No noteworthy findings were observed in the bone marrow specimens of the study drug administration group. In peripheral blood, as shown in Table 8, the number of red blood cells, hemoglobin,
No statistically significant differences were observed in hematocrit and platelets between the control group and the control group.

It is known that the number of white blood cells decreases with many anticancer drugs due to their bone marrow toxicity, but with these compounds, on the contrary, an increase or a tendency to increase was observed.

These results indicate that the indeno(1,2-Q) pyrazole derivative used as the active ingredient of the anticancer agent of the present invention has extremely low or no bone marrow toxicity.

In carrying out the present invention, it is administered in oral dosage forms such as tablets, capsules, granules, and syrups, or in parenteral dosage forms such as injections and suppositories, using commonly used formulation techniques. The dosage varies depending on the patient's condition, but for adults it is usually 20 -1 per day when administered orally.
1. In the case of parenteral administration, good results can be obtained at 2 to -300 μm.

Examples of formulations are given below.

Example 1 1-Methyl-3-(4-hydroxyphenyl)-indenoCL2-C)pyrazol-4NH)-one (compound number +7) 50kg Lactose +00klJ Crystalline cellulose 35kg Magnesium stearate 5kp Hydroxypropylmethylcellulose 7.5 kg Polyethylene Glycol 1500 1ktt titanium oxide
1.5kg Total 200kg The above ingredients are compressed into tablets and coated to make each tablet weigh 200m (contains 50m of active ingredient)
Coated tablets are manufactured.

Example 2 1-Methyl-3-(4-methoxyphenyl)-inz
(j, '2-c) Pyrazol-4(IH)-one (compound number +9) ' 25kg Magnesium stearate 51v Lactose 135kg Potato starch 50kl? Total weight: 250 kg The above mixture is granulated by dry granulation or wet granulation to produce granules containing 10% crude drug.

Example 5 One granule obtained according to Example 2 was placed in a No. 1 hard capsule.
Each capsule is filled with 250 rng to prepare capsules containing 25 my of the active ingredient per capsule.

Example 4 Sodium chloride 1602 Distilled water for injection Total amount 201 The above components are dissolved using a portion of distilled water for injection and the total amount is made up to 206 with distilled water for injection. After allowing sterilization with a 022μm filter, fill it into a 1ml ampoule, 1208C
The mixture was sterilized in an autoclave for 20 minutes to prepare a 1 me chain injection containing 10 mg of the active ingredient.

Example 5 1-Methyl-ro-(4-bromphenyl)-indeno[j,2-Q)pyrazole-4(IH
)-one (compound number +6) 200y was dissolved, purified soybean lecithin L2 kg and glycerol 2.5. kg
Add and warm to 40-80℃.

Add an appropriate amount of distilled water for injection, emulsify with a homomixer, and emulsify again with a high-pressure emulsifier. Distilled water for injection was added to bring the total volume to 1004, and the mixture was sterilized and filtered using a membrane filter. 5m
By filling e into a tube bottle, sealing it with a rubber stopper, and sterilizing it with high-pressure steam, it will contain no particles larger than 1 μm and have an average particle size of about 0.
An injectable emulsion having a diameter of 2 μm and containing 10 mg of the active ingredient is prepared.

Claims (1)

Scope of Claims: (1) General formula (wherein R1 is a hydrogen atom or a methyl group, R2 is a phenyl group or a pyridyl group with or without a substituent, and X is a carbonyl group or a substituent or Indeno (1
, 2-c) An anticancer agent containing a pyrazole derivative as an active ingredient. (2) A patent claim in which R2 is a phenyl group, a pyridyl group, a lower alkyl group having 1 or 2 carbon atoms, a phenyl group substituted with a halogen atom, a lower alkoxy group, a hydroxyl group, or an amine 7 group, and X is a carbonyl group range 1
The anticancer agent described in Section 1. (5) The anticancer agent according to claim 1, wherein x is a methylene group, a hydroxymethylene group, an aminomethylene group, or a lower alkylaminomethylene group. (4) The anticancer agent according to claim 1, which is in an oral administration form. (5) The anticancer agent according to claim 1, which is in a parenteral administration form.