JPH0643306B2 - Cancer cell metastasis inhibitor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial application] The present invention relates to an agent for suppressing cancer cell metastasis by the anti-metastatic effect of cancer cells of N-substituted-1-deoxynojirimycin derivatives.

[Conventional technology]

Most of the conventional anti-cancer agents are mainly composed of an active substance that kills malignant cells through the cell-killing property of the active substance or the human immune system. Many drugs of this type are currently studied and some of them are clinically used. However, it is still not sufficient for the treatment of cancer. In addition, solid cancers are often treated by surgery or radiation. It is expected that the effectiveness of these current various therapies will be further enhanced by suppressing the metastasis of cancer cells, but there are few substances whose main activity is anti-metastasis,
There is nothing clinically used.

[Problems to be Solved by the Invention]

It is an object of the present invention to find a substance that remarkably suppresses metastasis of cancer cells used for effective and appropriate treatment of cancer, and to provide a cancer cell metastasis inhibitor containing this as an active ingredient. .

[Means for Solving the Problems]

Cancer metastasis is a process in which (a) cancer cells are released from the primary focus, (b) disseminated in the blood vessel, (c) stagnated in capillaries, blocked, etc. and reach the target tissue to start proliferation. is there.

The present inventors have set up an experimental evaluation system for evaluating the suppressive action of this cancer metastasis, and based on this experimental evaluation system, the following general formula (1)
The present invention has been completed by finding that the N-substituted-1-deoxynojirimycin derivative represented by the formula (5) has an extremely excellent cancer cell metastasis inhibitory action.

The present invention has the general formula (I) [Wherein A represents a straight-chain or branched-chain hydrocarbon group having 3 to 5 carbon atoms with or without a double bond, R ₁ and R ₂ are the same or different hydrogen atoms, lower alkyl groups, Or a halogen atom] is a cancer cell metastasis inhibitor containing an N-substituted-1-deoxynojirimycin derivative represented by the following formula as an active ingredient.

The N-substituted-1-deoxynojirimycin represented by the general formula (I), which is an active ingredient of the present invention, is prepared by reacting 1-deoxynojirimycin with sodium hydrogencarbonate and aralkyl halide in dimethylformamide (DMF). , Can be obtained by isolation from the reaction solution (Japanese Patent Publication No. 55-9
051, gazette, Japanese Patent Publication No. 55-47655 gazette). In addition, 1-deoxynojirimycin, which is a raw material of these, was prepared by the present inventors from nojirimycin (5-
Amino-5-deoxy-D-glucopyranose) (see Japanese Patent Publication No. 43-760) is a known compound (Tetrahedron, 24, 2125-2144, 1968).
Year).

It is known that this N-substituted-1-deoxynojirimycin derivative has a blood glucose elevation suppressing action (see Japanese Patent Publication No. 55-9051 and Japanese Patent Publication No. 55-47655). However, it is not known that this substance has a cancer cell metastasis suppressing action.

Examples of the N-substituted-1-deoxynojirimycin derivative which is the active ingredient of the present invention include the following compounds. However, it is not limited to these.

N- (3-phenylpropyl) -1-deoxynojirimycin, N- (4-phenylbutyl) -1-deoxynojirimycin, N- (3-phenylbutyl) -1-deoxynojirimycin, N- (5- Phenylpentyl)-
1-deoxynojirimycin, N- (3-phenyl-2
-Propenyl) -1-deoxynojirimycin, N-
(4-phenyl-3-butenyl) -1-deoxynojirimycin, N- (3-methyl-3-phenyl-2-propenyl) -1-deoxynojirimycin, N- (3-m
-Methylphenyl-2-propenyl) -1-deoxynojirimycin, N- (3-p-methylphenyl-2-propenyl) -1-deoxynojirimycin, N- (3-
o-chlorophenyl-2-propenyl) -1-deoxynojirimycin, N- (3-p-chlorophenyl-3-
Methyl-2-propenyl) -1-deoxynojirimycin, N- (3-p-methylphenyl-3-methyl-2-
Propenyl) -1-deoxynojirimycin, N- (3
-P-Bromophenyl-2-propenyl) -1-deoxynojirimycin.

The cancer cell metastasis inhibitor of the present invention is an oral or parenteral preparation containing the above-mentioned N-substituted-1-deoxynojirimycin, and clinically passes through veins, arteries, skin, subcutaneous, intradermal, rectal and intramuscular, Or, it is administered orally. Further, a stronger effect can be expected by direct administration to the tumor. The dose varies depending on the administration form, dosage form or age, weight, and condition of the patient, but generally 100 to 3000 mg is administered once or several times a day.

As a parenteral preparation, a sterile aqueous or non-aqueous solution or an emulsion can be mentioned. Examples of the base of a non-aqueous solution or emulsion include propylene glycol, polyethylene glycol, glycerin, olive oil, corn oil, ethyl oleate and the like. Examples of oral preparations include capsules, tablets, granules, fine granules and powders. Starch, lactose, mannitol, ethyl cellulose, sodium carboxymethyl cellulose and the like are mixed as excipients in these preparations, and magnesium stearate or calcium stearate is added as a lubricant.
As the binder, gelatin, gum arabic, cellulose ester, polyvinylpyrrolidone or the like is used.

In addition, N-substituted-1-deoxynojirimycin derivatives N- (3-phenylpropyl) -1-deoxynojirimycin and N- (3-phenyl-2-propenyl) -1- which are the active ingredients of the present invention. The acute toxicity (LD ₅₀ value) in mice of deoxynojirimycin is both 0.5 g / kg or more by ip administration.

Next, formulation examples and effects of the present invention will be described.

[Example 1] N- (3-p-bromophenyl-2-propenyl) -1
-Deoxynojirimycin 50 mg Lactose 130 mg Potato starch 70 mg Polyvinylpyrrolidone 10 mg Magnesium stearate 2.5 mg Mix lactose and potato starch, add 20% ethanol solution of polyvinylpyrrolidone to it and wet evenly, and pass through a 1 mm mesh sieve. Dry at 45 ° C and repeat 1
It was passed through a mesh screen of mm. The granules thus obtained were mixed with magnesium stearate and molded into tablets.

[Example 2] N- (5-phenylpentyl) -1-deoxynojirimycin was crushed with a pin mill until it became particles of about 10 µm, and then 500 mg thereof was filled in a vial for injection. Separately Tween
805 mg and Japanese Pharmacopoeia 10 mg purified distilled water for injection 5 m
Dissolve in to make a solution. At the time of use, 5 m of this solution is added to N- (5-phenylpentyl) -1-deoxynojirimycin filled in the above vial to give a well-mixed suspension, and an appropriate amount is intramuscularly administered.

Efficacy test Test method From mouse tumor melanoma B16 strain Fiddler (F
idler) method (Method in Cancer Research, Volume 15, 399-
439, 1978), and a B16 high-metastatic strain was selected and used.

The metastatic inhibitory effect was evaluated based on the method of Kijima-Suda et al. (Cancer Research, 46, 858-862, 1986).

First, the B16 high-metastatic strain was planted in a Dulbecco's medium supplemented with fetal bovine serum, N-substituted-1-deoxynojirimycin represented by the general formula (I) was added, and then in the presence of 5% CO ₂ for _{2 to} 4 days. Cells grown at 37 ° C and grown were trypsin and EDTA.
The solution was peeled from the culture vessel. The cells were suspended in a balanced salt solution as living cells at 1 × 10 ⁶ cells per 1 m.

0.1 m of this suspension was transplanted from the tail vein of a mouse, and after breeding for 14 days, the abdomen was opened and the lung was excised. The colonies of the B16 high metastatic strain formed on the surface and inside the lung were counted and treated with a drug. Compared to the control, which was not.

Test Example 1 Cytotoxicity of N- (3-phenylpropyl) -1-deoxynojirimycin 10% of B16 high metastatic strain and mouse-derived tumor cell L929 strain
Dulbecco's medium supplemented with fetal bovine serum in the presence of 5% CO ₂ .
After culturing at 37 ° C, the cells were detached from the culture vessel with trypsin and EDTA solution.
10 ⁴ cells and L929 strain were suspended in the above medium so as to be 1 × 10 ³ cells. 150 μ of this suspension is added to N- (3-
50 μl of phenylpropyl) -1-deoxynojirimycin or adriamycin (control) solution was added and mixed. This was cultured for 3 days for the B16 high metastatic strain and for 4 days for the L929 strain, and the viability was determined under an inverted microscope to determine the cytotoxicity. The results are shown in Table 1.

As is clear from the above test results, 100 μg of N- (3-phenylpropyl) -1-deoxynojirimycin, which is the active ingredient of the present invention, was added to the B16 high-transgenic strain and the L929 strain.
No significant cytotoxicity was exhibited even at a high concentration of / m.

Test Example 2 Anti-metastatic effect of N- (3-phenylpropyl) -1-deoxynojirimycin The B16 high-metastatic strain was planted in a Dulbecco's medium containing 10% fetal bovine serum, and N-3-phenylpropyl) -1-deoxy was added. Add nojirimycin 30,100, or 300μg per 1m, 5
Cultured at 37 ° C in the presence of% CO ₂ for 3 days. The cells were peeled from the culture vessel in the same manner as in Test Example 1, and the cells were suspended in a balanced salt solution so that the number of viable cells was 1 × 10 ⁶ cells / m.
The 0.1 m was administered to the tail vein of a BDF ₁ mouse (8-week-old, male) and transplanted. After 14 days of breeding observation, the abdomen was opened and the lungs were excised, and the colonies of the B16 high metastatic strain formed on the surface and inside the lungs were counted. The results are shown in Table 2.

As is clear from the above test results, the number of colonies formed in the lung of N- (3-phenylpropyl) -1-deoxynojirimycin, which is the active ingredient of the present invention, decreased as the drug concentration was increased. , Lung metastasis is strongly suppressed.

Test Example 3 N- (3-phenyl-2-propenyl) -1
-Anti-metastatic action of deoxynojirimycin and N- (3-p-chlorophenyl-3-methyl-2-propenyl) -1-deoxynojirimycin N- (3-phenylpropyl) -1-in Test Example 2
Instead of deoxynojirimycin, N- (3-phenyl-2-propenyl) -1-deoxynojirimycin,
Using N- (3-p-chlorophenyl-3-methyl-2-propenyl) -1-deoxynojirimycin, the anti-metastatic effect was examined in the same manner as in Test Example 2. However, in this example, the B16 high-metastatic strain was transplanted with 2.5 × 10 ⁵ cells per mouse, 3 in each group. The results are shown in Table 3.

As is apparent from the above test results, N- (3-phenyl-2-propenyl) -1-deoxynojirimycin and N- (3-p-chlorophenyl-), which are the active ingredients of the present invention, are used.
3-Methyl-2-propenyl) -1-deoxynojirimycin remarkably suppressed the lung metastasis of the B16 high metastatic strain.

〔The invention's effect〕

The cancer cell metastasis inhibitor of the present invention is a drug that remarkably suppresses the diffusion and metastasis of cancer cells and has very little cytotoxicity. Therefore, when administered at the same time as the existing anticancer drug,
It is an extremely useful drug that can remarkably enhance the carcinostatic effect when used during surgery or radiation therapy.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Akira Nakabayashi 760 Shimooka-cho, Kohoku-ku, Yokohama-shi, Kanagawa Meiji Confectionery Co., Ltd. Pharmaceutical Research Laboratory (72) Inventor Takashi Tsuruoka 760, Shimooka-cho, Kohoku-ku, Yokohama-shi, Kanagawa Meiji Seika Co., Ltd., Pharmaceutical Research Laboratory (72) Inventor Shigeharu Inoue, 760, Shimooka-cho, Kohoku-ku, Yokohama-shi, Kanagawa Meiji Seika Co., Ltd., Pharmaceutical Research Laboratory (72) Inventor, Shinichi Kondo 760, Shimooka-cho, Kohoku-ku, Yokohama-shi, Kanagawa Prefecture Address Meiji Seika Co., Ltd. Pharmaceutical Research Center

Claims (1)

[Claims] 1. A general formula [In the formula, A represents a linear or branched hydrocarbon group having 3 to 5 carbon atoms, which may or may not have a double bond, and R ₁ , R ₂
Represent the same or different hydrogen atom, lower alkyl group, or halogen atom]. A cancer cell metastasis inhibitor comprising an N-substituted-1-deoxynojirimycin derivative represented by