JPH078793B2 - Cancer cell metastasis inhibitor containing N-substituted-1-deoxynojirimycin derivative - Google Patents

Description

TECHNICAL FIELD The present invention relates to an agent for suppressing cancer cell metastasis due to 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, and they are used in clinical trials. There is nothing that is said.

[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 by using this experimental evaluation system, the following general formula (I)
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 [Wherein X represents a linear or branched hydrocarbon group having a carbon number of 3 to 5 having or not having a double bond, Y represents oxygen or sulfur, R ₁ and R ₂ are the same or Represents a different hydrogen atom, a lower alkyl group, or a halogen atom] N-
It is a cancer cell metastasis inhibitor containing a substituted-1-deoxynojirimycin derivative 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 using 1-deoxynojirimycin as a raw material and a base such as sodium hydrogencarbonate in a polar solvent such as dimethylformamide (DMF). It can be obtained by reacting a lower aralkyl halide and isolating it from the reaction solution (see JP-A-60-166616). 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 therapeutic effect on diabetes, hyperlipidemia and steatosis (JP-A-60-166616).
(See Japanese Patent Publication). 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- (2-phenoxyethyl) -1-deoxynojirimycin, N- (3-phenoxypropyl) -1-deoxynojirimycin, N- (3-thiophenoxypropyl)-
1-deoxynojirimycin, N- (4-phenoxybutyl) -1-deoxynojirimycin, N- (5-phenoxypenkel-1-deoxynojirimycin, N- (3-phenoxy-trans-propen-1-yl) ) -1-Deoxynojirimycin, N- (3-phenoxy-trans-
2-propenyl) -1-deoxynojirimycin, N-
(4-phenoxy-trans-2-butenyl) -1-deoxynojirimycin, N- (4-thiophenoxy-trans-2-butenyl) -1-deoxynojirimycin, N
-(3-m-Methylphenoxypropyl) -1-deoxynojirimycin, N- (3-o-methylphenoxypropyl) -1-deoxynojirimycin, N- (3-p-methylphenoxypropyl) -1-deoxy Nojirimycin, N- (3-p-chlorphenoxy-3-methyl-2-
Propenyl) -1-deoxynojirimycin, N- (3-
p-methylphenoxy-3-methyl-2-propenyl)
-1-deoxynojirimycin, N- (3-p-chlorphenoxy-2-propenyl) -1-deoxynojirimycin, N- (3-m-fluorophenoxy-2-propenyl) -1-deoxynojirimycin, N -(3-p-Bromophenoxy-2-propenyl) -1-deoxynojirimycin, N- (4-p-chlorphenoxy-trans-
2-butenyl) -1-deoxynojirimycin.

The cancer cell metastasis inhibitor of the present invention is clinically administered as an oral or parenteral preparation containing the above-mentioned N-substituted-1-deoxynojirimycin via vein, artery, skin, subcutaneous, intradermal, rectal and intramuscular. , Or 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.

The acute toxicity in mice of N- (4-phenoxy-trans-2-butenyl) -1-deoxynojirimycin, which is the N-substituted-1-deoxynojirimycin derivative which is the active ingredient of the present invention, was intraperitoneally administered. When I checked in, LD
_{The 50} value was 0.5 g / kg or more.

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

[Example 1] N- (3-phenoxypropyl) -1-deoxynojirimycin 50 mg Lactose 130 mg Potato starch 70 mg Polyvinylpyrrolidone 10 mg Magnesium stearate 2.5 mg N- (3-phenoxypropyl) -1-deoxynojirimycin, lactose and Potato starch was mixed, and a 20% ethanol solution of polyvinylpyrrolidone was added to the mixture to uniformly wet it. The mixture was passed through a 1 mm mesh sieve, dried at 45 ° C., and passed through a 1 mm mesh sieve again. The granules thus obtained were mixed with magnesium stearate and molded into tablets.

[Example 2] N- (3-m-methylphenoxypropyl) -1-deoxynojirimycin was crushed with a pin mill until it became particles of about 10 µ, and then 500 mg thereof was filled in an injection vial. Separately, dissolve 5 mg of Tween 80 and 10 mg of purified JP in 10 ml of distilled water for injection to obtain a solution. At the time of use, 5 ml of this solution was added to N- (3-m-methylphenoxypropyl) -1-deoxynojirimycin filled in the previous vial.
Add l to mix well to form a suspension and administer an appropriate amount intramuscularly.

Efficacy test Test method From the melanoma B16 strain, which is a tumor cell of mouse, the method of Fidler (Method in Cancer Research, 15
Vol. 399-439, 1978) based on the selection of B16 high metastatic strains,
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 ME medium (DME medium) supplemented with fetal bovine serum, N-substituted-1-deoxynojirimycin represented by the general formula (I) was added, and the mixture was kept at 5% for 2-4 days.
After culturing at 37 ° C. in the presence of CO ₂ , the grown cells were detached from the culture vessel with a trypsin-EDTA solution. Call this cell Ca ⁺⁺
As live cells in Dulbecco's balanced salt solution without Mg ⁺⁺
The cells were suspended at 1 × 10 ⁶ cells / ml.

0.1 ml of this suspension was injected into the tail vein of the mouse, and after breeding for 14 days, the laparotomy was performed to remove the lung, and the metastatic nodules of the B16 high metastatic strain formed on the lung surface and inside were counted. Compared to untreated control.

Test Example 1 N- (4-phenoxy-trans-2-butenyl) -1-deoxynojirimycin cytotoxicity B16 high metastatic strain and mouse-derived tumor cell L929 strain in DME medium supplemented with 10% fetal calf serum After culturing at 37 ° C in the presence of 5% CO ₂ , the cells were detached from the culture vessel with a trypsin-EDTA solution, and 1 ml of B16 high metastatic strain became 1 × 10 ⁴ cells and L929 strain became 1 × 10 ³ cells per 1 ml. As above was suspended in the above medium. 150 μ of this suspension was added to N- (4-phenoxy-trans-
2-butenyl) -1-deoxynojirimycin or adriamycin (control) solution (50 μm) was added to each and mixed. The B16 high metastatic strain was cultured for 3 days and the L929 strain was cultured for 4 days, and life and death were observed under an inverted microscope to determine cytotoxicity. The results are shown in Table 1.

As is clear from the above test results, N- (4-phenoxy-trans-2-butenyl) which is the active ingredient of the present invention.
-1-Deoxynojirimycin is a highly metastatic strain of B16 and L929
Even at 30 μg / ml against the strain, no significant cytotoxicity was shown.

Test Example 2 Anti-metastatic effect of N- (4-phenoxy-trans-2-butenyl) -1-deoxynojirimycin The B16 high-metastatic strain was planted in a DME medium supplemented with 10% fetal calf serum,
N- (4-phenoxy-trans-2-butenyl) -1
-Add 30 μg of deoxynojirimycin per ml, 5
Incubated at 37 ° C in the presence of% CO ₂ for 3 days. The cells were detached from the culture vessel in the same manner as in Test Example 1 and suspended in a Dulbecco's balanced salt solution containing no Ca ⁺⁺ and Mg ⁺⁺ to give 1 × 10 ⁶ viable cells per ml, and 0.1 ml thereof was added. Was injected into the tail vein of BDF ₁ mouse (8 weeks old, male) to transplant cells. After 14 days of breeding observation, the abdomen was opened and the lungs were excised, and the metastatic nodules of the B16 high metastatic strain formed on the lung surface and inside were counted. The results are shown in Table 2.

As is clear from the above test results, N- (4-phenoxy-trans-2-butenyl) which is the active ingredient of the present invention.
In the test group of -1-deoxynojirimycin, the number of lung metastatic nodules was small, and metastasis was strongly suppressed.

Test Example 3 N- (2-phenoxyethyl) -1-deoxynojirimycin anti-metastatic action In Test Example 2, N- (4-phenoxy-trans-2).
N- (2-phenoxyethyl) -1-deoxynojirimycin was used instead of -butenyl) -1-deoxynojirimycin, and the anti-metastatic effect was examined in the same manner as in Test Example 2. The results are shown in Table 3.

As is clear from the above test results, metastasis of the B16 high metastatic strain to the lung was significantly suppressed by N- (2-phenoxyethyl) -1-deoxynojirimycin, which is the active ingredient of the present invention.

〔The invention's effect〕

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

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

Claims (1)

[Claims] 1. A general formula [Wherein X represents a straight-chain or branched hydrocarbon group having 3 to 5 carbon atoms with or without a double bond, Y represents oxygen or sulfur, and R ₁ and R ₂ are the same or Representing a different hydrogen atom, lower alkyl group or halogen atom] N
A cancer cell metastasis inhibitor containing a -substituted-1-deoxynojirimycin derivative as an active ingredient.