JPH078794B2 - Cancer cell metastasis inhibitor containing sugar lactam - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial field of use] The present invention relates to an agent for suppressing cancer cell metastasis due to the anti-metastatic effect of D-curcaro-δ-lactam or its alkyl ester group on cancer cells.

[Conventional technology]

The conventional anticancer agents have been mainly agents having 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 action is antimetastasis, and they are used in clinical trials. There is nothing that is said.

[Problems to be Solved by the Invention] The present invention has found a substance that significantly suppresses metastasis of cancer cells used for effective and appropriate treatment of cancer, and provides a cancer cell metastasis inhibitor containing the substance as an active ingredient The purpose is to do.

[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 discovering that the D-glucaro-δ-lactam or the alkyl ester group thereof represented by the formula (3) has an extremely excellent cancer cell metastasis inhibitory action.

The present invention has the general formula [Wherein R represents a hydrogen atom or a linear or branched alkyl group having 1 to 8 carbon atoms] or D-glucaro-δ-lactam or a pharmacologically acceptable salt or D- thereof.
It is a cancer cell metastasis inhibitor containing glucaro-δ-lactam alkyl ester as an active ingredient.

The compound represented by formula (I), which is the active ingredient of the present invention, is known. Regarding the method for producing D-glucaro-δ-lactam, the present inventors have found that the fermentation metabolite of actinomycetes, nojirimycin (5-amino-5-deoxy-D-glucopyranose), (Tetrahedron, 23, 2125, 1968) 3 by chemical or enzymatic oxidation.
That is, chemical oxidation of nojirimycin or treatment with glucose oxidase gives an oxidant of the 1-position hydroxyl group (D
-Gluco-δ-lactam), which is subsequently synthesized by catalytic air oxidation reaction (Annual Report of Meiji Seika, 13, 80)
~ See page 84, Japanese Examined Patent Publication No. 45-28375).

D-glucaro-δ-lactam alkyl ester is D-
It is obtained by reacting glucaro-δ-lactam with an alkyl halide or a cyazoalkane in methanol, tetrahydrofuran, dioxane, N, N-dimethylformamide, or a mixed solvent thereof (JP-A-56).
-34589 and Japanese Patent Publication No. 56-34590).

All of these are β-glucuronidase inhibitors (Japanese Patent Publication
45-28375, Japanese Patent Publication No. 56-34590, J. Biochem, 72 ,
207-211, 1972), and its evaluation as an anti-inflammatory agent (filed by the present inventors as Japanese Patent Application No. 62-90899) has already been determined. However, it is not known that this substance has a cancer cell inhibitory action.

The following compounds may be mentioned as active ingredients of the present invention. However, it is not limited to these.

D-Glucaro-δ-lactam, D-Glucaro-δ-lactam methyl ester, D-Glucaro-δ-lactam ethyl ester, D-Glucaro-δ-lactam-n-propyl ester, D-Glucaro-δ-lactam-n -Butyl ester.

Examples of pharmacologically acceptable salts of the compound of the present invention include ammonium salts, alkali metal salts such as sodium and potassium, alkaline earth metal salts such as magnesium and calcium, triethylamine, triethanolamine,
Examples thereof include salts with organic bases such as diethylaminoethylamine, salts with heterocyclic amines such as piperidine, piperazine and morpholine, or salts with amino acids such as lysine.

The cancer cell metastasis inhibitor of the present invention is the above-mentioned D-glocaro-δ-
Clinically intravenously as an oral or parenteral preparation containing lactam or D-glucaro-δ-lactam alkyl ester,
It is administered via arteries, 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.

Examples of parenteral preparations include sterile aqueous or non-aqueous solutions or emulsions. 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.

Regarding the active ingredient of the present invention, LD of acute toxicity in mice
_{The 50} value is D-glucaro-δ-lactam sodium salt is 3g / kg or more by intravenous injection, 5g / kg or more by oral administration, and D-glucaro-δ-lactam methyl ester is 5g / kg or more by intravenous injection, oral administration Was 10 g / kg or more.

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

〔Example〕

Example 1 D-Glucaro-δ-lactam potassium salt 50 mg Lactose 130 mg Potato starch 70 mg Polyvinylpyrrolidone 10 mg Magnesium stearate 2.5 mg D-Glucaro-δ-lactam potassium salt, lactose and potato starch were mixed, and 20% of polyvinylpyrrolidone was added thereto. % Ethanol solution was added to uniformly wet the mixture, and 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 D-Glucaro-δ-lactam ethyl ester 200 mg Lactose 50 mg CMC-calcium 100 mg Magnesium stearate 3 mg Hard gelatin capsules were filled with the above composition to prepare capsules.

Example 3 D-glucaro-δ-lactam sodium salt 50 mg / ml
The solution was dissolved in distilled water for injection so that the concentration became, and 5 ml of each ampoule was aseptically dispensed to encapsulate the ampoule.

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 Dulbecco's ME medium (DME medium) supplemented with fetal bovine serum, and D-glucaro-δ-lactam or D-glucaro-δ-lactam alkyl ester represented by the general formula (I) was added. , 2-4 days, in the presence of 5% CO ₂ 37
After culturing at 0 ° C., the grown cells were detached from the culture vessel with a trypsin-EDTA solution. The cells were treated with Dulbecco's balanced salt solution containing no Ca ⁺⁺ and Mg ⁺⁺ as living cells at 1 × 1 per ml.
The cells were suspended to give 0 ⁶ cells.

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 D-Glucaro-δ-lactam calcium salt cytotoxicity B16 high metastatic strain and mouse-derived tumor cell L929 strain in DME medium supplemented with 10% fetal bovine serum at 37 ° C. in the presence of 5% CO _2. After culturing, the cells were detached from the culture vessel with a trypsin-EDTA solution, and 1 ml of B16 high metastatic strain was 1 × 10 ⁴ cells and 1 of L929 strain was 1 ml.
The cells were suspended in the above medium to give 10 ³ cells. 150 μ of this suspension was added to 50 μ of each D-glucaro-δ-lactam calcium salt or adriamycin (control) solution and mixed. For B16 high metastatic strain, this was L929 for 3 days.
The strain was cultured for 4 days, and life and death was observed under an inverted microscope to determine cytotoxicity. The results are shown in Table 1.

As is clear from the above test results, the active ingredient of the present invention, D-glucaro-δ-lactam calcium salt, shows remarkable cytotoxicity against B16 highly metastatic strain and L929 strain even at a high concentration of 30 μg / ml. There wasn't.

Test Example 2 D-Glucaro-δ-lactam calcium salt transposition effect B16 high-transition strain was planted in DME medium containing 10% fetal calf serum,
D-glucaro-δ-lactam calcium salt 1 per ml
0 μg was added, and the mixture was cultured at 37 ° C. for 3 days in the presence of 5% CO ₂ . Remove cells from the culture vessel in the same manner as in Test Example 1
Suspend in 1 × 10 ⁶ viable cells in Dulbecco's balanced salt solution containing no Ca ⁺⁺ and Mg ^++, and add 0.1 ml of this to B
DF ₁ mice (8 weeks old, male) were injected into the tail vein and cells were transplanted. 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, the number of metastatic nodules formed in the lung was greatly reduced by the treatment with the D-glucaro-δ-lactam calcium salt, which is the active ingredient of the present invention.

Test Example 3 Anti-metastatic Action of D-Glucaro-δ-lactam Methyl Ester and D-Glucaro-δ-lactam Ethyl Ester In Test Example 2, instead of the D-glucaro-δ-lactam calcium salt, D-glucaro-δ-lactam. The anti-metastatic effect was examined in the same manner as in Test Example 2 using methyl ester and D-glucaro-δ-lactam ethyl ester. The results are shown in Table 3.

As is clear from the above test results, D-glucaro-δ-lactam methyl ester and D which are the active ingredients of the present invention.
-Glucaro-δ-lactam ethyl ester significantly suppressed the lung metastasis of B16 high metastatic strains.

〔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 R represents a hydrogen atom or a linear or branched alkyl group having 1 to 8 carbon atoms] or a D-glucaro-δ-lactam or a pharmacologically acceptable salt or D- thereof.
A cancer cell metastasis inhibitor containing glucaro-δ-lactam alkyl ester as an active ingredient.