JPS635026B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel bis-dioxopiperazine derivative and an antitumor agent comprising the same. The bis-dioxopiperazine derivative of the present invention has the following general formula () (In the formula, R ₁ represents a lower alkyl group, and R ₂ and R ₃ represent a saturated 6-membered heterocycle combined with adjacent nitrogen atoms. However, R ₂ and R ₃ represent the constituent atoms of the heterocycle. Contains any one of nitrogen, oxygen, or sulfur atoms in addition to the adjacent nitrogen atom, but in the case of a nitrogen atom, the heterocycle may be substituted with a lower alkyl group or a phenyl group. be. Various bis-dioxopiperazine derivatives have been reported to date, but 1,2-bis(4-morpholinomethyl-3.5-dioxopiperazin-1-yl) is the closest compound to the compound of the present invention.
- Ethane is known, and clinical effects have been reported as an antitumor agent and radiosensitizer (Proceedings of the 8th International Conference on Pharmacology, p. 441, 1981). The present inventor focused on the antitumor activity of this compound,
As a result of conducting research on more excellent derivatives of the same, the present invention was completed by discovering that the compound represented by the above general formula () has a wide range of antitumor activity and metastasis suppressing activity. The compound of the present invention is represented by the general formula (),
The symbol R ₁ in the formula represents a lower alkyl group. Examples of the lower alkyl group include straight-chain or branched-chain groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl and the like, with methyl being particularly preferred. R ₂ and R ₃ combine with adjacent nitrogen atoms to form a saturated 6
Indicates the formation of a membered heterocycle. However, the constituent atoms of the heterocycle include one of nitrogen, oxygen, or sulfur atoms in addition to the adjacent nitrogen atoms of R ₂ and R ₃ , but in the case of a nitrogen atom, the heterocycle is substituted with a lower alkyl group or a phenyl group. It means that it is okay to do something. Examples of the heterocycle formed by combining R ₂ and R ₃ with adjacent nitrogen atoms include piperazine, morpholine, thiomorpholine, etc. Substituents thereof include methyl, ethyl, n-propyl, iso-
Examples include lower alkyl groups such as propyl, and aryl groups such as phenyl and substituted phenyl. In addition, in the compound of general formula (), since the carbon atom to which R ₁ is bonded is an asymmetric carbon atom, not only the dl form (racemic form) but also the d form and l form optical isomers are included in the present invention. Included in compounds. Examples of the compounds of the present invention include the compounds described below. Γ dl of 1-(4-morpholinomethyl-3.5-dioxopiperazin-1-yl)-2-(3.5-dioxopiperazin-1-yl)-propane
body, d body and l body. Γ 1-(4-thiomorpholinomethyl3.5-dioxopiperazin-1-yl)-2-(3.5-dioxopiperazin-1-yl)-propane dl
body, d body and l body. Γ 1-[4-(N-phenylpiperazinomethyl)-3.5-dioxopiperazin-1-yl]
-2-(3.5-dioxopiperazin-1-yl)
- dl, d-form and l-form of propane. Γ 1-[4-(N-methylpiperazinomethyl)
-3.5-dioxopiperazin-1-yl]-2
-(3.5-dioxopiperazin-1-yl)-
dl, d and l forms of propane. The compound () of the present invention can be produced by the method shown below. That is, as the manufacturing method of the present invention, (A) general formula () (In the formula, R ₁ , R ₂ , R ₃ are the same as defined above) A method for obtaining the compound () of the present invention by solvolysis with a protic polar solvent, or (B) General formula() (In the formula, R ₁ is the same as the above definition.) Formaldehyde and the general formula ()

A method for obtaining the compound () of the present invention by reacting an amine represented by the formula: (wherein R ₂ and R ₃ are the same as defined above) is exemplified. Examples of protic polar solvents in method (A) include:
Examples include lower alcohols such as methanol, ethanol, n-butanol, and water. The reaction temperature of method (A) is -20 to 60°C, preferably 0 to 60°C.
It can be carried out in the range of 35℃, and the reaction time is 0.5
~5 hours is appropriate. Further, as a reaction solvent, for example, N,N-dimethylformamide (DMF),
Aprotic polar solvents such as dimethyl sulfoxide (DMSO), pyridine, ethyl acetate, chloroform, etc. may be combined with the protic polar solvents. (B) The amines in the general formula () are R ₂ and R ₃
N-methylpiperazine formed by combining, N
- Examples include cyclic amines such as phenylpiperazine, morpholine, and thiomorpholine. In the reaction of method (B), 0.8 to 1.1 mol of formaldehyde and the amine of general formula () are each used per 1 mol of the compound of general formula (). The reaction temperature of method (B) is -20 to 120℃, preferably 0
The reaction can be carried out at a temperature of -30°C, and the reaction time varies depending on the reaction temperature, but is in the range of 0.5 to 40 hours. Reaction solvents include DMF, DMSO, acetonitrile, ethyl acetate, methanol, ethanol,
Polar solvents such as butanol can be used alone or in combination. The compound of general formula (), which is the starting material for the method (A), is a new compound, and as shown in the production example, the compound of general formula () is reacted with at least twice the mole of formaldehyde and the amine of general formula (), respectively. It can be manufactured by Furthermore, the compound of general formula (), which is the starting material for the method (B), is a known compound and is disclosed in British Patent No. 1234935.
It can be produced according to the method described in the specification. Next, the bis-
The antitumor effect and toxicity of dioxopiperazine derivatives will be explained. The antitumor effect of the compounds of the present invention is shown in Lewis lung cancer, B.
- Confirmed by growth inhibition tests for malignant melanoma 16 and colon 38, as well as growth inhibition rates or survival prolongation rates for other syngeneic experimental tumors. In a growth inhibition test for Lewis Lung carcinoma, a group of 7 BDF was administered subcutaneously in the inguinal region of male mice (6 weeks old, weight 25 ± 2 g) at 5 × 10 ⁵
Lewis lung cancer cells were transplanted, and the test compound was orally administered in a prescribed amount once a day for 8 consecutive days starting from the next day. On the 20th day after tumor transplantation, the tumor was removed and its weight was measured. Summer. The effect of the test compound was expressed as growth inhibition rate (GI) using the following formula. Growth inhibition rate (%) = CT/C x 100 T: Average tumor weight of administration group C: Average tumor weight of control group The results are shown in Table 1 below.

[Table] The compound of the present invention showed a significantly higher growth inhibition rate against Lewis lung cancer cells than the comparative compound. It was also confirmed that lung metastasis of Lewis lung cancer was clearly suppressed compared to the control group. A growth inhibition test for B-16 malignant melanoma was performed by subcutaneously injecting 5 x 10 ⁵ B-16 malignant melanomas into the inguinal region of a group of 7 BDF and male mice (6 weeks old, weight 25 ± 2 g). The cells were transplanted, and the test compound was orally administered once a day for 8 consecutive days from the next day, and the tumor was removed on the 20th day after tumor transplantation.
This was done by measuring the weight. The effect of the test compound was determined as growth inhibition rate (Gl) in the same manner as the Lewis lung cancer growth inhibition test. The results are shown in Table 2 below.

[Table] The compound of the present invention showed a superior growth inhibition rate to B-16 malignant melanoma than the comparative compound. The growth inhibition test for Colon 38 is
Group of 7 BDF, male mice (6 weeks old, body weight 25±
2g), 40mg of Colon 38 tumor piece was implanted subcutaneously in the groin using a transplantation needle, and from the next day, the test compound was orally administered once a day for 8 consecutive days, and 30mg after tumor implantation.
The tumor was excised on the day of the experiment and its weight was measured. The effect of the test compound is the growth inhibition rate (G.
I.) was determined in the same manner as the Lewis lung cancer growth inhibition test. The results are shown in Table 3 below.

【table】

[Table] The compound of the present invention showed a growth inhibition rate equivalent to or better than that of the comparative compound against Colon 38. In the survival test for P388 leukemia tumor, 1.0 × 10 ⁶ P388 leukemia tumor cells were each intraperitoneally transplanted into a group of 10 CDF male mice (6 weeks old, body weight 25 ± 2 g). The experiment was carried out by intraperitoneally administering a predetermined amount of the test compound for 9 consecutive days. The effect of the test compound is the increase of life
Span: ILS) was calculated using the following formula. Survival prolongation rate (%) = (T/C-1) x 100 T: Average survival days of the administration group C: Average survival days of the control group The results are shown in Table 4 below.

[Table] The compound of the present invention showed an excellent survival rate against P388 leukemia tumor cells. Next, the acute toxicity of the compound of the present invention was confirmed by the following test. The acute toxicity test was conducted using a group of 10 ddY male mice (5
The test compound prepared in physiological saline supplemented with 0.5% carboxymethyl cellulose (CMC) was administered intraperitoneally to mice (age, body weight 23 ± 2 g) and observed for 14 days. -Wilcoxon) method
Asked for _LD50 . The results are shown in Table 5 below.

[Table] Next, the administration method, dosage form, and dosage when the bis-dioxopiperazine derivative of the present invention is applied to the human body will be explained. The compound of the present invention can be administered orally or parenterally, and dosage forms for oral administration include tablets, coatings, powders, granules, capsules, syrups, etc.; can be used as injections (including freeze-dried injections that are dissolved before use), suppositories, etc. The preparation of these dosage forms may include pharmaceutically acceptable excipients, binders, lubricants, disintegrants, suspending agents, emulsifiers, preservatives, stabilizers and dispersants, such as lactose, sucrose, starch. , dextrin, crystalline cellulose, kaolin, calcium carbonate, talc, magnesium stearate,
It is carried out using distilled water or physiological saline. The dosage varies depending on the patient's symptoms, age, weight, etc., but the daily dose for adults is 50 to 3000.
mg, preferably 500 to 1000 mg, can be administered in 1 to 3 divided doses. As mentioned above, the compound () of the present invention is a compound of the known 1,2-bis(4-morpholinomethyl-3.5-
Not only can dioxopiperazin-1-yl)-ethane be expected to have antitumor effects and radiosensitizing effects, but the test results show that it has a broader antitumor spectrum than the known comparative compounds.
Since it has been confirmed that antitumor activity is enhanced, it is extremely useful as an antitumor agent. Next, production examples will be shown to explain the present invention in more detail, but the present invention is not limited thereto. Production example 1 dl-1,2-bis(4-morpholinomethyl-
3.5-dioxopiperazin-1-yl)-propane dl-1,2-bis(3,5-dioxopiperazin-1-yl)-propane 2.0g (7.4m mol) and morpholine 1.4ml (16.2m mol) After adding 20 ml of DMF and 5 ml of absolute ethanol and stirring at 150°C for 10 minutes,
1.40 ml of a 37% formaldehyde aqueous solution was added, and the mixture was heated and stirred at the same temperature for an additional 1.5 hours. After the reaction is complete,
The reaction mixture was heated, and the liquid was concentrated under reduced pressure to obtain crystals, which were thoroughly washed with ether and dried under reduced pressure to obtain 2.36 g (yield: 67.8%) of the title intermediate compound. Melting point: 163-165℃ Elemental analysis value: _{C21H34N6O6 Theoretical} value (%): C: _54.06 H: _7.35 N: 18.01 Actual value (%): C: 54.28 H: 7.58 N: 18.05 Manufacture The following intermediate compounds were prepared from the corresponding starting materials in the same manner as in Example 1. Γ dl-1.2-bis(4-thiomorpholinomethyl-3.5-dioxopiperazin-1-yl)-propane Melting point: 140-143℃ Elemental analysis value: Theoretical value as C ₂₁ H ₃₄ N ₆ S ₂ O ₄ (% ):
C: 50.58 H: 6.87 N: 16.85 S: 12.86 Actual value (%):
C: 50.33 H: 6.95 N: 16.71 S: 12.66 Γdl-1.2-bis[4-(N-phenylpiverazinomethyl)-3.5-dioxopiperazine-1-
] - Propane Melting point: 98-101℃ Elemental analysis value: C ₃₃ H ₄₄ N ₈ O ₄ Theoretical value (%): C: 64.26 H: 7.19 N: 18.17 Actual value (%): C: 64.10 H: 7.28 N: 18.02 Production Example 2 dl-1-(4-monopholinomethyl-3.5-dioxopiperazin-1-yl)-2-(3.5-dioxopiperazin-1-yl)-propane Intermediate compound obtained in Production Example 1 above Add 200 ml of water to 4.0 g of dl-1.2-bis(4-morpholinomethyl-3.5-dioxopiperazin-1-yl)-propane, stir at room temperature for 0.5 hours, then separate the suspended matter and stir the liquid for another 3 hours at room temperature. did. After freezing the reaction mixture in a dry ice-acetone bath, the temperature of this frozen product is gradually raised at room temperature to obtain an aqueous solution (liquid temperature 0).
The colorless solid precipitated in the water (℃) was collected and dried over phosphorus pentoxide under reduced pressure to give 1.22 g of the title compound (yield: 40
%) was obtained. Melting point: 162-165℃ Elemental analysis value: Theoretical value (%) as C ₁₆ H ₂₅ N ₅ O ₅ : C: 52.31 H: 6.86 N: 19.06 Actual value (%): C: 52.05 H: 6.95 N: 19.26 Red External absorption spectrum (KBr) cm ^-1 : 2960, 2850, 2800, 1700, 1685

【table】 / \
Mass spectrum: m/e=367 (M ⁺ ) Production example 3 dl-1-(4-thiomorpholinomethyl-3.5-
Dioxopiperazin-1-yl)-2-
(3.5-dioxopiperazin-1-yl)-propane Intermediate compound dl-1,2-bis(4-thiomorpholinomethyl-3.5-dioxopiperazin-1-yl)-propane obtained in Production Example 1 0.6 g water 30
ml to form a suspension solution, which was stirred at room temperature for 5 hours. After the reaction was completed, the reaction mixture was filtered, and the white precipitate collected was dried under reduced pressure. Add 1 ml of chloroform to this white precipitate and stir at room temperature for 20 minutes.
Insoluble matter was removed and dried under reduced pressure to obtain 0.2 g (yield 43%) of the title compound. Melting point: 205.5-208.5℃ Elemental analysis value: Theoretical value (%) as C ₁₆ H ₂₅ N ₅ SO ₄ :
C: 50.12 H: 6.57 N: 18.26 S: 8.36 Actual value (%):
C50.41 H: 6.40 N: 18.01 S: 8.16 Infrared absorption spectrum (KBr) cm ^-1 : 2960, 2800, 1700, 1680

【table】 / \
The following compound was produced from the corresponding intermediate compound in the same manner as in Production Example 3. Γ dl-1-[4-(N-phenylpiperazinomethyl)-3.5-dioxopiperazin-1-yl]
-2-(3.5-dioxopiperazin-1-yl)
-Propane melting point: 188-190℃ Elemental analysis value: Theoretical value (%) as C ₂₂ H ₃₀ N ₆ O ₄ : C: 59.71 H: 6.83 N: 18.99 Actual value (%): C: 59.50 H: 6.99 N: 18.85 Infrared absorption spectrum (KBr) cm ^-1 : 2975, 2820, 1710, 1690

【table】

[Table] Production example 4 dl-1-[4-(N-phenylpiperazinomethyl)-3.5-dioxopiperazin-1-yl]
-2-(3.5-dioxopiperazin-1-yl)
-Propane dl-1.2-bis(3.5-dioxopiperazine-1
-yl)-propane 1.34g (51m mol), N-phenylpiperazine 0.81g (5m mol), DMF 13.4
ml, absolute ethanol 3.3ml, formaldehyde
0.43 ml of 37% aqueous solution was added and stirred at room temperature for 48 hours.
After the reaction was completed, a white precipitate was collected, washed successively with absolute ethanol and ether, and then dried under reduced pressure to obtain 1.39 g (yield: 62%) of the title compound. It was confirmed from the melting point (188-190°C) and infrared absorption spectrum that the title compound obtained was the same as the compound obtained in the same manner as Production Example 3.

Claims (1)

[Claims] 1 General formula () (In the formula, R ₁ represents a lower alkyl group, and R ₂ and R ₃ represent a saturated 6-membered heterocycle combined with adjacent nitrogen atoms. However, R ₂ and R ₃ represent the constituent atoms of the heterocycle. contains any one of nitrogen, oxygen, or sulfur atoms in addition to the adjacent nitrogen atom, and in the case of a nitrogen atom, the heterocycle may be substituted with a lower alkyl group or a phenyl group. Dioxopiperazine derivative. 2 General formula () (In the formula, R ₁ represents a lower alkyl group, and R ₂ and R ₃ represent a saturated 6-membered heterocycle combined with adjacent nitrogen atoms. However, R ₂ and R ₃ represent the constituent atoms of the heterocycle. contains any one of nitrogen, oxygen, or sulfur atoms in addition to the adjacent nitrogen atom, and in the case of a nitrogen atom, the heterocycle may be substituted with a lower alkyl group or a phenyl group. An antitumor agent characterized by containing a dioxopiperazine derivative as an active ingredient.